Sample records for por sds-page nivel

  1. Preparative SDS PAGE as an Alternative to His-Tag Purification of Recombinant Amelogenin

    PubMed Central

    Gabe, Claire M.; Brookes, Steven J.; Kirkham, Jennifer

    2017-01-01

    Recombinant protein technology provides an invaluable source of proteins for use in structure-function studies, as immunogens, and in the development of therapeutics. Recombinant proteins are typically engineered with “tags” that allow the protein to be purified from crude host cell extracts using affinity based chromatography techniques. Amelogenin is the principal component of the developing enamel matrix and a frequent focus for biomineralization researchers. Several groups have reported the successful production of recombinant amelogenins but the production of recombinant amelogenin free of any tags, and at single band purity on silver stained SDS PAGE is technically challenging. This is important, as rigorous structure-function research frequently demands a high degree of protein purity and fidelity of protein sequence. Our aim was to generate His-tagged recombinant amelogenin at single band purity on silver stained SDS PAGE for use in functionality studies after His-tag cleavage. An acetic acid extraction technique (previously reported to produce recombinant amelogenin at 95% purity directly from E. coli) followed by repeated rounds of nickel column affinity chromatography, failed to generate recombinant amelogenin at single band purity. This was because following an initial round of nickel column affinity chromatography, subsequent cleavage of the His-tag was not 100% efficient. A second round of nickel column affinity chromatography, used in attempts to separate the cleaved His-tag free recombinant from uncleaved His-tagged contaminants, was still unsatisfactory as cleaved recombinant amelogenin exhibited significant affinity for the nickel column. To solve this problem, we used preparative SDS PAGE to successfully purify cleaved recombinant amelogenins to single band purity on silver stained SDS PAGE. The resolving power of preparative SDS PAGE was such that His-tag based purification of recombinant amelogenin becomes redundant. We suggest that acetic

  2. Determination of the Subunit Molecular Mass and Composition of Alcohol Dehydrogenase by SDS-PAGE

    ERIC Educational Resources Information Center

    Nash, Barbara T.

    2007-01-01

    SDS-PAGE is a simple, rapid technique that has many uses in biochemistry and is readily adaptable to the undergraduate laboratory. It is, however, a technique prone to several types of procedural pitfalls. This article describes the use of SDS-PAGE to determine the subunit molecular mass and composition of yeast alcohol dehydrogenase employing…

  3. Identification of fish species after cooking by SDS-PAGE and urea IEF: a collaborative study.

    PubMed

    Etienne, M; Jérôme, M; Fleurence, J; Rehbein, H; Kündiger, R; Mendes, R; Costa, H; Pérez-Martín, R; Piñeiro-González, C; Craig, A; Mackie, I; Malmheden Yman, I; Ferm, M; Martínez, I; Jessen, F; Smelt, A; Luten, J

    2000-07-01

    A collaborative study, to validate the use of SDS-PAGE and urea IEF, for the identification of fish species after cooking has been performed by nine laboratories. By following optimized standard operation procedures, 10 commercially important species (Atlantic salmon, sea trout, rainbow trout, turbot, Alaska pollock, pollack, pink salmon, Arctic char, chum salmon, and New Zealand hake) had to be identified by comparison with 22 reference samples. Some differences in the recoveries of proteins from cooked fish flesh were noted between the urea and the SDS extraction procedures used. Generally, the urea extraction procedure appears to be less efficient than the SDS extraction for protein solubilization. Except for some species belonging to the Salmonidae family (Salmo, Oncorhynchus), both of the analytical techniques tested (urea IEF, SDS-PAGE) enabled identification of the species of the samples to be established. With urea IEF, two laboratories could not differentiate Salmo salar from Salmo trutta. The same difficulties were noted for differentiation between Oncorhynchus gorbuscha and Oncorhynchus keta samples. With SDS-PAGE, three laboratories had some difficulties in identifying the S. trutta samples. However, in the contrast with the previous technique, SDS-PAGE allows the characterization of most of the Oncorhynchus species tested. Only Oncorhynchus mykiss was not clearly recognized by one laboratory. Therefore, SDS-PAGE (Excel gel homogeneous 15%) appears to be better for the identification, after cooking, of fish such as the tuna and salmon species which are characterized by neutral and basic protein bands, and urea IEF (CleanGel) is better for the gadoid species, which are characterized by acid protein bands (parvalbumins). Nevertheless, in contentious cases it is preferable to use both analytical methods.

  4. Teaching the structure of immunoglobulins by molecular visualization and SDS-PAGE analysis.

    PubMed

    Rižner, Tea Lanišnik

    2014-01-01

    This laboratory class combines molecular visualization and laboratory experimentation to teach the structure of the immunoglobulins (Ig). In the first part of the class, the three-dimensional structures of the human IgG and IgM molecules available through the RCSB PDB database are visualized using freely available software. In the second part, IgG and IgM are studied using electrophoretic methods. Through SDS-PAGE analysis under reducing conditions, the students determine the number and molecular masses of the polypeptide chains, while through SDS-PAGE under nonreducing conditions, the students assess the oligomerization of these Ig molecules. The aims of this class are to expand upon the knowledge and understanding of the Ig structure that the students have gained from classroom lectures. The combination of this molecular visualization of the Ig molecules and the SDS-PAGE experimentation ensures variety in the teaching techniques, while the implication of the Ig molecules in human disease promotes interest for biomedical students. © 2014 by The International Union of Biochemistry and Molecular Biology.

  5. Real-time Fluorescence Polarization Microscopy of the Moving Boundary in Cross-Gradient SDS-PAGE

    NASA Astrophysics Data System (ADS)

    Hwang, Jeeseong; Giulian, Gary

    2003-03-01

    Real-time Fluorescence Polarization Microscopy of the Moving Boundary in Cross-Gradient SDS-PAGE Jeeseong Hwang, Jeffrey R. Krogmeier, Angela M. Bardo, Scott N. Goldie, Lori S. Goldner; Optical Technology Division, National Institute of Standards and Technology, Gaithersburg, MD 20899 Gary G. Giulian, Carl R. Merril; National Institute of Mental Health, National Institutes of Health, Bethesda, MD 20892 Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE) is a popular method to separate proteins by their apparent molecular weight. However, it is a limited technique due, in part, to its low spatial resolution. In order to improve the resolution and to enhance the detection sensitivity of proteins separated by SDS-PAGE we are studying the detergent properties at the moving boundary of precast Tris-Tricine-Acetate cross-gradient gels using fluorescent cationic and pH indicating dyes. We have developed real-time full-field fluorescence polarization microscopy to monitor the dynamic fluorescence anisotropy from the cationic tetramethylindocarbocyanine dyes localized in the "extended stack", a concentrated detergent zone. We will present quantitative results of the fluorescence anisotropy. Our system is capable of analyzing local structures of the detergent molecules in the moving boundary of SDS-PAGE and the microenvironment(s) near the boundary. We will discuss the significance of these results and their potential role in enhanced protein separation.

  6. Effects of high intensity exercise on isoelectric profiles and SDS-PAGE mobility of erythropoietin.

    PubMed

    Voss, S; Lüdke, A; Romberg, S; Schänzer, E; Flenker, U; deMarees, M; Achtzehn, S; Mester, J; Schänzer, W

    2010-06-01

    Exercise induced proteinuria is a common phenomenon in high performance sports. Based on the appearance of so called "effort urines" in routine doping analysis the purpose of this study was to investigate the influence of exercise induced proteinuria on IEF profiles and SDS-PAGE relative mobility values (rMVs) of endogenous human erythropoietin (EPO). Twenty healthy subjects performed cycle-ergometer exercise until exhaustion. VO (2)max, blood lactate, urinary proteins and urinary creatinine were analysed to evaluate the exercise performance and proteinuria. IEF and SDS-PAGE analyses were performed to test for differences in electrophoretic behaviour of the endogenous EPO before and after exercise. All subjects showed increased levels of protein/creatinine ratio after performance (8.8+/-5.2-26.1+/-14.4). IEF analysis demonstrated an elevation of the relative amount of basic band areas (13.9+/-11.3-36.4+/-12.6). Using SDS-PAGE analysis we observed a decrease in rMVs after exercise and no shift in direction of the recombinant human EPO (rhEPO) region (0.543+/-0.013-0.535+/-0.012). Following identification criteria of the World Anti Doping Agency (WADA) all samples were negative. The implementation of the SDS-PAGE method represents a good solution to distinguish between results influenced by so called effort urines and results of rhEPO abuse. Thus this method can be used to confirm adverse analytical findings.

  7. SDS-PAGE analysis of Aβ oligomers is disserving research into Alzheimer´s disease: appealing for ESI-IM-MS

    NASA Astrophysics Data System (ADS)

    Pujol-Pina, Rosa; Vilaprinyó-Pascual, Sílvia; Mazzucato, Roberta; Arcella, Annalisa; Vilaseca, Marta; Orozco, Modesto; Carulla, Natàlia

    2015-10-01

    The characterization of amyloid-beta peptide (Aβ) oligomer forms and structures is crucial to the advancement in the field of Alzheimer´s disease (AD). Here we report a critical evaluation of two methods used for this purpose, namely sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), extensively used in the field, and ion mobility coupled to electrospray ionization mass spectrometry (ESI-IM-MS), an emerging technique with great potential for oligomer characterization. To evaluate their performance, we first obtained pure cross-linked Aβ40 and Aβ42 oligomers of well-defined order. Analysis of these samples by SDS-PAGE revealed that SDS affects the oligomerization state of Aβ42 oligomers, thus providing flawed information on their order and distribution. In contrast, ESI-IM-MS provided accurate information, while also reported on the chemical nature and on the structure of the oligomers. Our findings have important implications as they challenge scientific paradigms in the AD field built upon SDS-PAGE characterization of Aβ oligomer samples.

  8. Genetic Variability among Lucerne Cultivars Based on Biochemical (SDS-PAGE) and Morphological Markers

    NASA Astrophysics Data System (ADS)

    Farshadfar, M.; Farshadfar, E.

    The present research was conducted to determine the genetic variability of 18 Lucerne cultivars, based on morphological and biochemical markers. The traits studied were plant height, tiller number, biomass, dry yield, dry yield/biomass, dry leaf/dry yield, macro and micro elements, crude protein, dry matter, crude fiber and ash percentage and SDS- PAGE in seed and leaf samples. Field experiments included 18 plots of two meter rows. Data based on morphological, chemical and SDS-PAGE markers were analyzed using SPSSWIN soft ware and the multivariate statistical procedures: cluster analysis (UPGMA), principal component. Analysis of analysis of variance and mean comparison for morphological traits reflected significant differences among genotypes. Genotype 13 and 15 had the greatest values for most traits. The Genotypic Coefficient of Variation (GCV), Phenotypic Coefficient of Variation (PCV) and Heritability (Hb) parameters for different characters raged from 12.49 to 26.58% for PCV, hence the GCV ranged from 6.84 to 18.84%. The greatest value of Hb was 0.94 for stem number. Lucerne genotypes could be classified, based on morphological traits, into four clusters and 94% of the variance among the genotypes was explained by two PCAs: Based on chemical traits they were classified into five groups and 73.492% of variance was explained by four principal components: Dry matter, protein, fiber, P, K, Na, Mg and Zn had higher variance. Genotypes based on the SDS-PAGE patterns all genotypes were classified into three clusters. The greatest genetic distance was between cultivar 10 and others, therefore they would be suitable parent in a breeding program.

  9. Analysis of the axial filaments of Treponema hyodysenteriae by SDS-PAGE and immunoblotting.

    PubMed

    Kent, K A; Sellwood, R; Lemcke, R M; Burrows, M R; Lysons, R J

    1989-06-01

    Purified axial filaments from eight serotypes of Treponema hyodysenteriae and two non-pathogenic intestinal spirochaetes were characterized by SDS-PAGE and Western blotting. Axial filaments of all ten strains had similar SDS-PAGE profiles; five major axial filament polypeptides were identified, with molecular masses of 43.8, 38, 34.8, 32.8 and 29.4 kDa. Hyperimmune gnotobiotic pig serum raised against purified axial filaments of strain P18A (serotype 4) cross-reacted with all other serotypes and with the non-pathogens, and convalescent serum taken from a pig with persistent swine dysentery also showed a strong response to the axial filament polypeptides. Hyperimmune gnotobiotic pig serum raised against axial filaments failed to agglutinate viable organisms and did not inhibit growth in vitro. Hence, the axial filaments of T. hyodysenteriae have been identified as major immunodominant antigens, although the role that antibodies to these antigens play in protection has yet to be established.

  10. Purification of recombinant Aβ(1-42) and pGlu-Aβ(3-42) using preparative SDS-PAGE.

    PubMed

    Spahn, Claudia; Wermann, Michael; Eichentopf, Rico; Hause, Gerd; Schlenzig, Dagmar; Schilling, Stephan

    2017-08-01

    Recombinant expression and purification of amyloid peptides represents a common basis for investigating the molecular mechanisms of amyloid formation and toxicity. However, the isolation of the recombinant peptides is hampered by inefficient separation from contaminants such as the fusion protein required for efficient expression in E. coli. Here, we present a new approach for the isolation of highly purified Aβ(1-42) and pGlu-Aβ(3-42), which is based on a separation using preparative SDS-PAGE. The method relies on the purification of the Aβ fusion protein by affinity chromatography followed by preparative SDS-PAGE under reducing conditions and subsequent removal of detergents by precipitation. The application of preparative SDS-PAGE represents the key step to isolate highly pure recombinant Aβ, which has been applied for characterization of aggregation and toxicity. Thereby, the yield of the purification strategy was  >60%. To the best of our knowledge, this is the first description of an electrophoresis-based method for purification of a recombinant Aβ peptide. Therefore, the method might be of interest for isolation of other amyloid peptides, which are critical for conventional purification strategies due to their aggregation propensity. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Lab-on-a-chip and SDS-PAGE analysis of hemolymph protein profile from Rhipicephalus microplus (Acari: Ixodidae) infected with entomopathogenic nematode and fungus.

    PubMed

    Golo, Patrícia Silva; Dos Santos, Alessa Siqueira de Oliveira; Monteiro, Caio Marcio Oliveira; Perinotto, Wendell Marcelo de Souza; Quinelato, Simone; Camargo, Mariana Guedes; de Sá, Fillipe Araujo; Angelo, Isabele da Costa; Martins, Marta Fonseca; Prata, Marcia Cristina de Azevedo; Bittencourt, Vânia Rita Elias Pinheiro

    2016-09-01

    In the present study, lab-on-a-chip electrophoresis (LoaC) was suggested as an alternative method to the conventional polyacrylamide gel electrophoresis under denaturing conditions (SDS-PAGE) to analyze raw cell-free tick hemolymph. Rhipicephalus microplus females were exposed to the entomopathogenic fungus Metarhizium anisopliae senso latu IBCB 116 strain and/or to the entomopathogenic nematode Heterorhabditis indica LPP1 strain. Hemolymph from not exposed or exposed ticks was collected 16 and 24 h after exposure and analyze by SDS-PAGE or LoaC. SDS-PAGE yielded 15 bands and LoaC electrophoresis 17 bands. Despite the differences in the number of bands, when the hemolymph protein profiles of exposed or unexposed ticks were compared in the same method, no suppressing or additional bands were detected among the treatments regardless the method (i.e., SDS-PAGE or chip electrophoresis using the Protein 230 Kit®). The potential of LoaC electrophoresis to detect protein bands from tick hemolymph was considered more efficient in comparison to the detection obtained using the traditional SDS-PAGE method, especially when it comes to protein subunits heavier than 100 KDa. LoaC electrophoresis provided a very good reproducibility, and is much faster than the conventional SDS-PAGE method, which requires several hours for one analysis. Despite both methods can be used to analyze tick hemolymph composition, LoaC was considered more suitable for cell-free hemolymph protein separation and detection. LoaC hemolymph band percent data reported changes in key proteins (i.e., HeLp and vitellogenin) exceptionally important for tick embryogenesis. This study reported, for the first time, tick hemolymph protein profile using LoaC.

  12. Combination of SDS-PAGE and intact mass analysis for rapid determination of heterogeneities in monoclonal antibody therapeutics.

    PubMed

    Yamada, Hideaki; Matsumura, Chiemi; Yamada, Keita; Teshima, Koichiro; Hiroshima, Takashi; Kinoshita, Mitsuhiro; Suzuki, Shigeo; Kakehi, Kazuaki

    2017-05-01

    mAbs are currently mainstream in biopharmaceuticals, and their market has been growing due to their high target specificity. Characterization of heterogeneities in mAbs is performed to secure their quality and safety by physicochemical analyses. However, they require time-consuming task, which often strain the resources of drug development in pharmaceuticals. Rapid and direct method to determine the heterogeneities should be a powerful tool for pharmaceutical analysis. Considering the advantages of electrophoresis and MS, this study addresses the combination of SDS-PAGE and intact mass analysis, which provides direct, rapid, and orthogonal determination of heterogeneities in mAb therapeutics. mAb therapeutics that migrated in SDS-PAGE were recovered from gel by treatment with SDC-containing buffer. Usage of SDC-containing buffer as extraction solvent and ethanol-based staining solution enhanced the recovery of intact IgG from SDS-PAGE gels. Recovery of mAbs reached more than 86% with 0.2% SD. The heterogeneities, especially N-glycan variants in the recovered mAb therapeutics, were clearly determined by intact mass analysis. We believe that the study is important in pharmaceuticals‧ perspective since orthogonal combination of gel electrophoresis and intact mass analysis should be pivotal role for rapid and precise characterization of mAbs. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. SDS-PAGE Electrophoretic Property of Human Chorionic Gonadotropin (hCG) and its β-subunit

    PubMed Central

    2005-01-01

    The microheterogeneity property of hCG with regards to its sialic acid contents resulted in variable mobility of the glycoprotein in SDS-PAGE. The intact hCG molecule is composed of two dissimilar subunits, namely α- and β-subunits. The identification of hCG bands in SDS-PAGE was accomplished by the immunoblotting experiment, whereby the antibody directed toward the specific region of β-subunit of hCG was used. The data shows that the different mobility of intact hCG was attributed to the different degree of desialylation of the glycoprotein. Nevertheless, unlike the intact hCG, the mobility of its β-subunit was not affected by its variety sialic acid content. This characteristic of β-hCG is beneficial when semi-quantification of total hCG is required. Quantification of hCG using the HPLC-reversed phase C18 analytical column is not possible as the glycoprotein was eluted in multiple fractions at different retention times. The identification of denatured hCG (HPLC eluted fractions) was carried out by immunoblotting experiment whilst immunoassay technique failed to detect its presence in any fraction. PMID:16094462

  14. Two-dimensional blue native/SDS-PAGE analysis of whole cell lysate protein complexes of rice in response to salt stress.

    PubMed

    Hashemi, Amenehsadat; Gharechahi, Javad; Nematzadeh, Ghorbanali; Shekari, Faezeh; Hosseini, Seyed Abdollah; Salekdeh, Ghasem Hosseini

    2016-08-01

    To understand the biology of a plant in response to stress, insight into protein-protein interactions, which almost define cell behavior, is thought to be crucial. Here, we provide a comparative complexomics analysis of leaf whole cell lysate of two rice genotypes with contrasting responses to salt using two-dimensional blue native/SDS-PAGE (2D-BN/SDS-PAGE). We aimed to identify changes in subunit composition and stoichiometry of protein complexes elicited by salt. Using mild detergent for protein complex solubilization, we were able to identify 9 protein assemblies as hetero-oligomeric and 30 as homo-oligomeric complexes. A total of 20 proteins were identified as monomers in the 2D-BN/SDS-PAGE gels. In addition to identifying known protein complexes that confirm the technical validity of our analysis, we were also able to discover novel protein-protein interactions. Interestingly, an interaction was detected for glycolytic enzymes enolase (ENO1) and triosephosphate isomerase (TPI) and also for a chlorophyll a-b binding protein and RuBisCo small subunit. To show changes in subunit composition and stoichiometry of protein assemblies during salt stress, the differential abundance of interacting proteins was compared between salt-treated and control plants. A detailed exploration of some of the protein complexes provided novel insight into the function, composition, stoichiometry and dynamics of known and previously uncharacterized protein complexes in response to salt stress. Copyright © 2016 Elsevier GmbH. All rights reserved.

  15. Protein identification and quantification from riverbank grape, Vitis riparia: Comparing SDS-PAGE and FASP-GPF techniques for shotgun proteomic analysis.

    PubMed

    George, Iniga S; Fennell, Anne Y; Haynes, Paul A

    2015-09-01

    Protein sample preparation optimisation is critical for establishing reproducible high throughput proteomic analysis. In this study, two different fractionation sample preparation techniques (in-gel digestion and in-solution digestion) for shotgun proteomics were used to quantitatively compare proteins identified in Vitis riparia leaf samples. The total number of proteins and peptides identified were compared between filter aided sample preparation (FASP) coupled with gas phase fractionation (GPF) and SDS-PAGE methods. There was a 24% increase in the total number of reproducibly identified proteins when FASP-GPF was used. FASP-GPF is more reproducible, less expensive and a better method than SDS-PAGE for shotgun proteomics of grapevine samples as it significantly increases protein identification across biological replicates. Total peptide and protein information from the two fractionation techniques is available in PRIDE with the identifier PXD001399 (http://proteomecentral.proteomexchange.org/dataset/PXD001399). © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. The rumen microbial metaproteome as revealed by SDS-PAGE.

    PubMed

    Snelling, Timothy J; Wallace, R John

    2017-01-07

    Ruminal digestion is carried out by large numbers of bacteria, archaea, protozoa and fungi. Understanding the microbiota is important because ruminal fermentation dictates the efficiency of feed utilisation by the animal and is also responsible for major emissions of the greenhouse gas, methane. Recent metagenomic and metatranscriptomic studies have helped to elucidate many features of the composition and activity of the microbiota. The metaproteome provides complementary information to these other -omics technologies. The aim of this study was to explore the metaproteome of bovine and ovine ruminal digesta using 2D SDS-PAGE. Digesta samples were taken via ruminal fistulae and by gastric intubation, or at slaughter, and stored in glycerol at -80 °C. A protein extraction protocol was developed to maximise yield and representativeness of the protein content. The proteome of ruminal digesta taken from dairy cows fed a high concentrate diet was dominated by a few very highly expressed proteins, which were identified by LC-MS/MS to be structural proteins, such as actin and α- and β-tubulins, derived from ciliate protozoa. Removal of protozoa from digesta before extraction of proteins revealed the prokaryotic metaproteome, which was dominated by enzymes involved in glycolysis, such as glyceraldehyde-3-phosphate dehydrogenase, phosphoenolpyruvate carboxykinase, phosphoglycerate kinase and triosephosphate isomerase. The enzymes were predominantly from the Firmicutes and Bacteroidetes phyla. Enzymes from methanogenic archaea were also abundant, consistent with the importance of methane formation in the rumen. Gels from samples from dairy cows fed a high proportion of grass silage were consistently obscured by co-staining of humic compounds. Samples from beef cattle and fattening lambs receiving a predominantly concentrate diet produced clearer gels, but the pattern of spots was inconsistent between samples, making comparisons difficult. This work demonstrated for the

  17. Comparison of low molecular weight glutenin subunits identified by SDS-PAGE, 2-DE, MALDI-TOF-MS and PCR in common wheat.

    PubMed

    Liu, Li; Ikeda, Tatsuya M; Branlard, Gerard; Peña, Roberto J; Rogers, William J; Lerner, Silvia E; Kolman, María A; Xia, Xianchun; Wang, Linhai; Ma, Wujun; Appels, Rudi; Yoshida, Hisashi; Wang, Aili; Yan, Yueming; He, Zhonghu

    2010-06-24

    Low-molecular-weight glutenin subunits (LMW-GS) play a crucial role in determining end-use quality of common wheat by influencing the viscoelastic properties of dough. Four different methods - sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), two-dimensional gel electrophoresis (2-DE, IEF x SDS-PAGE), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) and polymerase chain reaction (PCR), were used to characterize the LMW-GS composition in 103 cultivars from 12 countries. At the Glu-A3 locus, all seven alleles could be reliably identified by 2-DE and PCR. However, the alleles Glu-A3e and Glu-A3d could not be routinely distinguished from Glu-A3f and Glu-A3g, respectively, based on SDS-PAGE, and the allele Glu-A3a could not be differentiated from Glu-A3c by MALDI-TOF-MS. At the Glu-B3 locus, alleles Glu-B3a, Glu-B3b, Glu-B3c, Glu-B3g, Glu-B3h and Glu-B3j could be clearly identified by all four methods, whereas Glu-B3ab, Glu-B3ac, Glu-B3ad could only be identified by the 2-DE method. At the Glu-D3 locus, allelic identification was problematic for the electrophoresis based methods and PCR. MALDI-TOF-MS has the potential to reliably identify the Glu-D3 alleles. PCR is the simplest, most accurate, lowest cost, and therefore recommended method for identification of Glu-A3 and Glu-B3 alleles in breeding programs. A combination of methods was required to identify certain alleles, and would be especially useful when characterizing new alleles. A standard set of 30 cultivars for use in future studies was chosen to represent all LMW-GS allelic variants in the collection. Among them, Chinese Spring, Opata 85, Seri 82 and Pavon 76 were recommended as a core set for use in SDS-PAGE gels. Glu-D3c and Glu-D3e are the same allele. Two new alleles, namely, Glu-D3m in cultivar Darius, and Glu-D3n in Fengmai 27, were identified by 2-DE. Utilization of the suggested standard cultivar set, seed of which is available from the

  18. In vitro pepsin resistance of proteins: Effect of non-reduced SDS-PAGE analysis on fragment observation.

    PubMed

    Pickles, Juliette; Rafiq, Samera; Cochrane, Stella A; Lalljie, Anja

    2014-01-01

    The introduction of novel proteins to food products carries with it the need to assess the potential allergenicity of such materials. Resistance to in vitro pepsin digestion is one parameter considered in such a risk assessment using a weight of evidence approach; however, the methodology used to investigate this has not been fully standardised. In vitro pepsin resistance assays typically involve SDS-PAGE performed under reducing conditions, with limited published data available on the assay using non-reducing conditions despite the need to consider non-reducing analysis techniques having been highlighted by regulatory bodies such as the European Food Safety Authority (EFSA). The purpose of the work reported here was to investigate the applicability of (and additional insight provided by) non-reducing analyses, by digesting a set of proteins using a ring-trial validated method, with analysis by SDS-PAGE under both reducing and non-reducing conditions. In silico prediction of digest fragments was also investigated. Significant differences were observed between results under reduced and non-reduced conditions for proteins in which disulphide bonds have a major role in protein structure, such as ribulose 1,5-diphosphate carboxylase (RUBISCO) and bovine serum albumin. For proteins with no or few disulphide bonds, no significant differences were seen in the results. Structural information such as disulphide bond numbers and positions should be considered during experimental design, as a non-reduced approach may be appropriate for some proteins. The in silico approach was a useful tool to suggest potential digest fragments, however the predictions were not always confirmed in vitro and should be considered a guide only.

  19. A high precision method for length-based separation of carbon nanotubes using bio-conjugation, SDS-PAGE and silver staining.

    PubMed

    Borzooeian, Zahra; Taslim, Mohammad E; Ghasemi, Omid; Rezvani, Saina; Borzooeian, Giti; Nourbakhsh, Amirhasan

    2018-01-01

    Parametric separation of carbon nanotubes, especially based on their length is a challenge for a number of nano-tech researchers. We demonstrate a method to combine bio-conjugation, SDS-PAGE, and silver staining in order to separate carbon nanotubes on the basis of length. Egg-white lysozyme, conjugated covalently onto the single-walled carbon nanotubes surfaces using carbodiimide method. The proposed conjugation of a biomolecule onto the carbon nanotubes surfaces is a novel idea and a significant step forward for creating an indicator for length-based carbon nanotubes separation. The conjugation step was followed by SDS-PAGE and the nanotube fragments were precisely visualized using silver staining. This high precision, inexpensive, rapid and simple separation method obviates the need for centrifugation, additional chemical analyses, and expensive spectroscopic techniques such as Raman spectroscopy to visualize carbon nanotube bands. In this method, we measured the length of nanotubes using different image analysis techniques which is based on a simplified hydrodynamic model. The method has high precision and resolution and is effective in separating the nanotubes by length which would be a valuable quality control tool for the manufacture of carbon nanotubes of specific lengths in bulk quantities. To this end, we were also able to measure the carbon nanotubes of different length, produced from different sonication time intervals.

  20. Functionality screen of streptavidin mutants by non-denaturing SDS-PAGE using biotin-4-fluorescein.

    PubMed

    Humbert, Nicolas; Ward, Thomas R

    2008-01-01

    Site-directed mutagenesis or directed evolution of proteins often leads to the production of inactive mutants. For streptavidin and related proteins, mutations may lead to the loss of their biotin-binding properties. With high-throughput screening methodologies in mind, it is imperative to detect, prior to the high-density protein production, the bacteria that produce non-functional streptavidin isoforms. Based on the incorporation of biotin-4-fluorescein in streptavidin mutants present in Escherichia coli bacterial extracts, we detail a functional screen that allows the identification of biotin-binding streptavidin variants. Bacteria are cultivated in a small volume, followed by a rapid treatment of the cells; biotin-4-fluorescein is added to the bacterial extract and loaded on an Sodium Dodecyl Sulfate Poly-Acrylamide Gel Electrophoresis (SDS-PAGE) under non-denaturing conditions. Revealing is performed using a UV transilluminator. This screen is thus easy to implement, cheap and requires only readily available equipment.

  1. Comparative examination of adsorption of serum proteins on HSA- and PLGA-based nanoparticles using SDS-PAGE and LC-MS.

    PubMed

    Gossmann, R; Fahrländer, E; Hummel, M; Mulac, D; Brockmeyer, J; Langer, K

    2015-06-01

    The behavior of nanosized drug carrier systems under cell culture conditions and therefore also the destiny in the body are highly influenced by the protein corona, which is formed upon entering a biological environment. Some of the adsorbed proteins, named opsonins, lead to a shortened plasma circulation half-life of the nanoparticles. Others are attributed to promote the transport of nanoparticles into other compartments of the body, just to mention two examples. Hence, detailed knowledge concerning the composition of the protein corona is of great importance. The aim of this work was to investigate the influence of the nanoparticle starting material and the surface modification on the composition of the adsorbed serum proteins in a cell culture environment. Therefore, positively charged nanoparticles based on the biodegradable polymer poly(dl-lactide-co-glycolide) (PLGA) stabilized with didodecyldimethylammonium bromide (DMAB) and negatively charged nanoparticles based on human serum albumin (HSA) were prepared and modified with hydrophilic polymers. By incubating the nanoparticles with fetal bovine serum (FBS) the adsorption of serum proteins on the colloidal system was investigated. Using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) a semi-quantitative analysis of the protein corona was performed and after enzymatic in-solution-digestion the adsorbed proteins were identified using high resolution LC-MS. Our study accentuates the influence of the core material, surface charge, and surface modification on the amount and nature of the adsorbed proteins. The combination of SDS-PAGE and LC-MS turns out to be a simple and reliable method to investigate the protein corona of nanoparticles. Copyright © 2015 Elsevier B.V. All rights reserved.

  2. Characterization of glycoprotein biopharmaceutical products by Caliper LC90 CE-SDS gel technology.

    PubMed

    Chen, Grace; Ha, Sha; Rustandi, Richard R

    2013-01-01

    Over the last decade, science has greatly improved in the area of protein sizing and characterization. Efficient high-throughput methods are now available to substitute for the traditional labor-intensive SDS-PAGE methods, which alternatively take days to analyze a very limited number of samples. Currently, PerkinElmer(®) (Caliper) has designed an automated chip-based fluorescence detection method capable of analyzing proteins in minutes with sensitivity similar to standard SDS-PAGE. Here, we describe the use and implementation of this technology to characterize and screen a large number of formulations of target glycoproteins in the 14-200 kDa molecular weight range.

  3. Proteomic analysis of human bladder epithelial cells by 2D blue native SDS-PAGE reveals TCDD-induced alterations of calcium and iron homeostasis possibly mediated by nitric oxide.

    PubMed

    Verma, Nisha; Pink, Mario; Petrat, Frank; Rettenmeier, Albert W; Schmitz-Spanke, Simone

    2015-01-02

    A proteomic analysis of the interaction among multiprotein complexes involved in 2,3,7,8-dibenzo-p-dioxin (TCDD)-mediated toxicity in urinary bladder epithelial RT4 cells was performed using two-dimensional blue native SDS-PAGE (2D BN/SDS-PAGE). To enrich the protein complexes, unexposed and TCDD-exposed cells were fractionated. BN/SDS-PAGE of the resulting fractions led to an effective separation of proteins and protein complexes of various origins, including cell membrane, mitochondria, and other intracellular compartments. Major differences between the proteome of control and exposed cells involved the alteration of many calcium-regulated proteins (calmodulin, protein S100-A2, annexin A5, annexin A10, gelsolin isoform b) and iron-regulated proteins (ferritin, heme-binding protein 2, transferrin). On the basis of these findings, the intracellular calcium concentration was determined, revealing a significant increase after 24 h of exposure to TCDD. Moreover, the concentration of the labile iron pool (LIP) was also significantly elevated in TCDD-exposed cells. This increase was strongly inhibited by the calmodulin (CaM) antagonist W-7, which pointed toward a possible interaction between iron and calcium signaling. Because nitric oxide (NO) production was significantly enhanced in TCDD-exposed cells and was also inhibited by W-7, we hypothesize that alterations in calcium and iron homeostasis upon exposure to TCDD may be linked through NO generated by CaM-activated nitric oxide synthase. In our model, we propose that NO produced upon TCDD exposure interacts with the iron centers of iron-regulatory proteins (IRPs) that modulate the alteration of ferritin and transferrin, resulting in an augmented cellular LIP and, hence, increased toxicity.

  4. Accelerated SDS depletion from proteins by transmembrane electrophoresis: Impacts of Joule heating.

    PubMed

    Unterlander, Nicole; Doucette, Alan Austin

    2018-02-08

    SDS plays a key role in proteomics workflows, including protein extraction, solubilization and mass-based separations (e.g. SDS-PAGE, GELFrEE). However, SDS interferes with mass spectrometry and so it must be removed prior to analysis. We recently introduced an electrophoretic platform, termed transmembrane electrophoresis (TME), enabling extensive depletion of SDS from proteins in solution with exceptional protein yields. However, our prior TME runs required 1 h to complete, being limited by Joule heating which causes protein aggregation at higher operating currents. Here, we demonstrate effective strategies to maintain lower TME sample temperatures, permitting accelerated SDS depletion. Among these strategies, the use of a magnetic stir bar to continuously agitate a model protein system (BSA) allows SDS to be depleted below 100 ppm (>98% removal) within 10 min of TME operations, while maintaining exceptional protein recovery (>95%). Moreover, these modifications allow TME to operate without any user intervention, improving throughput and robustness of the approach. Through fits of our time-course SDS depletion curves to an exponential model, we calculate SDS depletion half-lives as low as 1.2 min. This promising electrophoretic platform should provide proteomics researchers with an effective purification strategy to enable MS characterization of SDS-containing proteins. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. An investigation into the use of SDS-PAGE of cell surface extracts and proteolytic activity to differentiate Prevotella nigrescens and Prevotella intermedia.

    PubMed

    Cookson, A L; Wray, A; Handley, P S; Jacob, A E

    1996-02-15

    By comparison of the cell surface proteins derived from the outer membrane and fibrils from 14 Prevotella intermedia and 19 Prevotella nigrescens strains using SDS and analysed by SDS-PAGE, it was possible to distinguish the two species. A polypeptide of approx. 21 kDa distinguished P. intermedia strains, whereas two polypeptides of approx. 18 and 22 kDa could be used to identify P. nigrescens strains. Four other human oral black pigmented bacterial species (Porphyromonas gingivalis, Prevotella denticola, Prevotella loescheii and Prevotella melaninogenica) did not have the 18-, 21- or 22-kDa polypeptides shown by P. intermedia or P. nigrescens. The cell-associated proteolytic activity of eight strains of P. intermedia, 14 strains of P. nigrescens and one strain of P. gingivalis (W50) was assessed using four chromogenic substrates. The hydrolysis of the substrate GPPNA (indicative of dipeptidyl peptidase IV-like activity) and SAAPPNA (elastase-like activity) by P. intermedia strains varied from 32 to 114 units and 0.5 to 12.6 units of activity respectively, where one unit was defined as the amount of protease enzyme catalysing the formation of 1 nmol of p-nitroaniline under experimental conditions. 37.5% (3 of 8) of P. intermedia strains hydrolysed SAAPPNA (chymotrypsin-like enzyme activity) with activities of between 7 and 12 units. The hydrolysis of GPPNA and SAAAPNA by P. nigrescens strains was 32-149 and 3-16 units, respectively. 57% (8 of 14) of P. nigrescens strains hydrolysed SAAPPPNA with activities ranging from 3 to 8 units. None of the P. intermedia or P. nigrescens strains examined were found to have trypsin-like enzyme activity (BAPNA hydrolysis). The GPPNA and SAAAPNA hydrolytic activity associated with the proteases from Porphyromonas gingivalis W50 was at least twice that of P. intermedia and P. nigrescens strains. The similar peptidase activities of P. intermedia and P. nigrescens against chromogenic substrates cannot be used to differentiate the

  6. Genetic variation of jointed goatgrass (Aegilops cylindrica Host.) from Iran using RAPD-PCR and SDS-PAGE of seed proteins.

    PubMed

    Farkhari, M; Naghavi, M R; Pyghambari, S A; Sabokdast

    2007-09-01

    Genetic variation of 28 populations of jointed goatgrass (Aegilops cylindrica Host.), collected from different parts of Iran, were evaluated using both RAPD-PCR and SDS-PAGE of seed proteins. The diversity within and between populations for the three-band High Molecular Weight (HMW) subunits of glutenin pattern were extremely low. Out of 15 screened primers of RAPD, 14 primers generated 133 reproducible fragments which among them 92 fragments were polymorphic (69%). Genetic similarity calculated from the RAPD data ranged from 0.64 to 0.98. A dendrogram was prepared on the basis of a similarity matrix using the UPGMA algorithm and separated the 28 populations into two groups. Confusion can happen between populations with the same origin as well as between populations of very diverse geographical origins. Our results show that compare to seed storage protein, RAPD is suitable for genetic diversity assessment in Ae. cylindrica populations.

  7. Using SDS-PAGE gel fingerprinting to identify soft-bodied wood-boring insect larvae to species.

    PubMed

    O'Neill, Mark A; Denos, Mia; Reed, Daniel

    2018-03-01

    This paper describes the progress that we have made in assessing the feasibility of 'fingerprinting' using imaged SDS-PAGE gels of haemolymph proteins, to identify soft-bodied wood-boring insect larvae such as the Asian longhorn beetle, Anoplophora glabripennis (Motscholsky, 1853) (Coleoptera: Cerambycidae). Because of stringent import restrictions and difficulty in obtaining licences to work with these organisms, we opted to work with four species of scarab beetle, Mecynorhina polyphemus (Fabricius, 1781), Pachnoda sinuata (Fabricius, 1775), Eucidella shiratica (Csiki, 1909) and Eucidella shultzeorum (Kolbe, 1906) which have near identical larval morphologies. We show that this technology when combined with an advanced pattern matching system (Digital Automated Identification SYstem - DAISY) can classify soft-bodied insect larvae that are almost identical morphologically to species at a level of accuracy is in excess of 98%. The study also indicates that the technology copes well with noisy data and small training sets. The experience gained in undertaking this study gives us confidence that we will be able to develop a field deployable system in the medium term. We believe that as a high-throughput identification tool, this technology is superior to competitor technologies (e.g. fingerprinting of imaged DNA gels) in terms of speed, cost and ease of use; and therefore, is suitable for low-cost deployment in the field. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  8. Development of Two Analytical Methods Based on Reverse Phase Chromatographic and SDS-PAGE Gel for Assessment of Deglycosylation Yield in N-Glycan Mapping.

    PubMed

    Eckard, Anahita D; Dupont, David R; Young, Johnie K

    2018-01-01

    N -lined glycosylation is one of the critical quality attributes (CQA) for biotherapeutics impacting the safety and activity of drug product. Changes in pattern and level of glycosylation can significantly alter the intrinsic properties of the product and, therefore, have to be monitored throughout its lifecycle. Therefore fast, precise, and unbiased N -glycan mapping assay is desired. To ensure these qualities, using analytical methods that evaluate completeness of deglycosylation is necessary. For quantification of deglycosylation yield, methods such as reduced liquid chromatography-mass spectrometry (LC-MS) and reduced capillary gel electrophoresis (CGE) have been commonly used. Here we present development of two additional methods to evaluate deglycosylation yield: one based on LC using reverse phase (RP) column and one based on reduced sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE gel) with offline software (GelAnalyzer). With the advent of rapid deglycosylation workflows in the market for N -glycan profiling replacing overnight incubation, we have aimed to quantify the level of deglycosylation in a selected rapid deglycosylation workflow. Our results have shown well resolved peaks of glycosylated and deglycosylated protein species with RP-LC method allowing simple quantification of deglycosylation yield of protein with high confidence. Additionally a good correlation, ≥0.94, was found between deglycosylation yields estimated by RP-LC method and that of reduced SDS-PAGE gel method with offline software. Evaluation of rapid deglycosylation protocol from GlycanAssure™ HyPerformance assay kit performed on fetuin and RNase B has shown complete deglycosylation within the recommended protocol time when evaluated with these techniques. Using this kit, N -glycans from NIST mAb were prepared in 1.4 hr and analyzed by hydrophilic interaction chromatography (HILIC) ultrahigh performance LC (UHPLC) equipped with a fluorescence detector (FLD

  9. A simple SDS-PAGE protein pattern from pitcher secretions as a new tool to distinguish Nepenthes species (Nepenthaceae).

    PubMed

    Biteau, Flore; Nisse, Estelle; Miguel, Sissi; Hannewald, Paul; Bazile, Vincent; Gaume, Laurence; Mignard, Benoit; Hehn, Alain; Bourgaud, Frederic

    2013-12-01

    Carnivorous plants have always fascinated scientists because these plants are able to attract, capture, and digest animal prey using their remarkable traps that contain digestive secretions. Nepenthes is one of the largest genera of carnivorous plants, with 120 species described thus far. Despite an outstanding diversity of trap designs, many species are often confused with each other and remain difficult to classify because they resemble pitchers or of the occurrence of interspecific hybrids. Here, we propose a new method to easily distinguish Nepenthes species based on a SDS PAGE protein pattern analysis of their pitcher secretions. Intraspecific comparisons were performed among specimens growing in different environmental conditions to ascertain the robustness of this method. Our results show that, at the juvenile stage and in the absence of prey in the pitcher, an examined species is characterized by a specific and stable profile, whatever the environmental conditions. The method we describe here can be used as a reliable tool to easily distinguish between Nepenthes species and to help with potential identification based on the species-specific protein pattern of their pitcher secretions, which is complementary to the monograph information.

  10. Nonelectrophoretic bidirectional transfer of a single SDS-PAGE gel with multiple antigens to obtain 12 immunoblots.

    PubMed

    Kurien, Biji T; Scofield, R Hal

    2009-01-01

    Protein blotting is an invaluable technique in immunology to detect and characterize proteins of low abundance. Proteins resolved on sodium dodecyl sulfate (SDS) polyacrylamide gels are normally transferred electrophoretically to adsorbent membranes such as nitrocellulose or polyvinylidene diflouride membranes. Here, we describe the nonelectrophroretic transfer of the Ro 60 (or SSA) autoantigen, 220- and 240-kD spectrin antigens, and prestained molecular weight standards from SDS polyacrylamide gels to obtain up to 12 immunoblots from a single gel and multiple sera.

  11. Fluorescent Labeling of Proteins and Its Application to SDS-PAGE and Western Blotting.

    PubMed

    Alba, F Javier; Bartolomé, Salvador; Bermúdez, Antonio; Daban, Joan-Ramon

    2015-01-01

    This chapter describes very simple fluorescent methods developed in our laboratory allowing the rapid monitoring of total protein patterns on both sodium dodecyl sulfate (SDS) polyacrylamide gels and western blots. The noncovalent dye Nile red (9-diethylamino-5H-benzo[α]phenoxazine-5-one) is used for the sensitive staining of proteins in SDS gels. This method is compatible with the electroblotting of protein bands and with the staining of the resulting blot with the covalent dye MDPF (2-methoxy-2,4-diphenyl-3(2H)-furanone). These staining procedures are applied sequentially; there is no need to run a duplicate unstained gel for protein blotting. Furthermore, since only the adduct formed by the reaction of MDPF with proteins is fluorescent, there is no need to destain the membrane after protein labeling. In addition, MDPF staining is compatible with further immunodetection of specific bands with polyclonal antibodies. Finally, using the adequate conditions described below, MDPF staining does not preclude the N-terminal sequence analysis of proteins in selected bands.

  12. Analysis by two-dimensional Blue Native/SDS-PAGE of membrane protein alterations in rat soleus muscle after hindlimb unloading.

    PubMed

    Basco, Davide; Nicchia, Grazia Paola; Desaphy, Jean-François; Camerino, Diana Conte; Frigeri, Antonio; Svelto, Maria

    2010-12-01

    Muscle atrophy occurring in several pathophysiological conditions determines decreases in muscle protein synthesis, increases in the rate of proteolysis and changes in muscle fiber composition. To determine the effect of muscle atrophy induced by hindlimb unloading (HU) on membrane proteins from rat soleus, a proteomic approach based on two-dimensional Blue Native/SDS-PAGE was performed. Proteomic analysis of normal and HU soleus muscle demonstrates statistically significant changes in the relative level of 36 proteins. Among the proteins identified by mass spectrometry, most are involved in pathways associated with muscle fuel utilization, indicating a shift in metabolism from oxidative to glycolytic. Moreover, immunoblotting analysis revealed an increase in aquaporin-4 (AQP4) water channel and an alteration of proteins belonging to the dystrophin-glycoprotein complex (DGC). AQP4 and DGC are regulated in soleus muscle subjected to simulated microgravity in response to compensatory mechanisms induced by muscle atrophy, and they parallel the slow-to-fast twitch conversion that occurs in soleus fibers during HU. In conclusion, the alterations of soleus muscle membrane proteome may play a pivotal role in the mechanisms involved in disuse-induced muscle atrophy.

  13. Hybridization parameters revisited: solutions containing SDS.

    PubMed

    Rose, Ken; Mason, John O; Lathe, Richard

    2002-07-01

    Salt concentration governs nucleic acid hybridization according to the Schildkraut-Lifson equation. High concentrations of SDS are used in some common protocols, but the effects of SDS on hybridization stringency have not been reported. We investigated hybridization parameters in solutions containing SDS. With targets immobilized on nylon membranes and PCR- or transcription-generated probes, we report that the 50% dissociation temperature (Tm*) in the absence of SDS was 15 degrees C-17degrees C lower than the calculated Tm. SDS had only modest effects on Tm* [1% (w/v) equating to 8 mM NaCl]. RNA/DNA hybrids were approximately 11 degrees C more stable than DNA/DNA hybrids. Incomplete homology (69%) significantly reduced the Tm* for DNA/DNA hybrids (approximately /4degrees C; 0.45 degrees C/% nonhomology) but far less so for RNA/DNA hybrids (approximately 2.3 degrees C; approximately 0.07 degrees C/% non-homology); incomplete homology also markedly reduced the extent of hybridization. On these nylonfilters, SDS had a major effect on nonspecific binding. Buffers lacking SDS, or with low salt concentration, gave high hybridization backgrounds; buffers containing SDS, or high-salt buffers, gave reproducibly low backgrounds.

  14. Resistance of bromelain to SDS binding.

    PubMed

    Bhattacharya, Reema; Bhattacharyya, Debasish

    2009-04-01

    Interaction of the plant cysteine protease bromelain with SDS has been studied using CD spectroscopy, intrinsic fluorescence emission, extrinsic fluorescence probe pyrene, isothermal calorimetric (ITC) investigations and inhibition of hydrolyzing activity. Results exhibit number of synchronous transitions when plotted against the total SDS concentration. SDS at submicellar level caused conformation change of bromelain leading to a stable entity. ITC and pyrene experiments suggest that the structural modifications below 5 mM, the cmc(app) of SDS solutions containing bromelain, are the result of alterations of solvent hydrophobicity or non-specific weak binding and/or adsorption of SDS monomers. Melting temperature (T(m)) and the free energy change for thermal unfolding (DeltaG(unf)) of the SDS induced conformers was decreased by 5 degrees C and 0.5 kcal/mol respectively, compared to native bromelain. Below 5 mM, SDS caused large decrease in V(max) without affecting K(m) for the substrate Z-Arg-Arg-NHMec. Analysis of kinetic data imply that SDS acts as a partial non-competitive inhibitor since even at 100 mM, the residual activity of bromelain was retained by 3%. Inhibition studies show an IC(50) of 0.55 mM and a high K(i) of 0.145 mM. These demonstrate that bromelain is resistant to SDS binding and denaturation, a property known for beta-sheet rich kinetically stable proteins.

  15. SDS: A Framework for Scientific Data Services

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Dong, Bin; Byna, Surendra; Wu, Kesheng

    2013-10-31

    Large-scale scientific applications typically write their data to parallel file systems with organizations designed to achieve fast write speeds. Analysis tasks frequently read the data in a pattern that is different from the write pattern, and therefore experience poor I/O performance. In this paper, we introduce a prototype framework for bridging the performance gap between write and read stages of data access from parallel file systems. We call this framework Scientific Data Services, or SDS for short. This initial implementation of SDS focuses on reorganizing previously written files into data layouts that benefit read patterns, and transparently directs read callsmore » to the reorganized data. SDS follows a client-server architecture. The SDS Server manages partial or full replicas of reorganized datasets and serves SDS Clients' requests for data. The current version of the SDS client library supports HDF5 programming interface for reading data. The client library intercepts HDF5 calls and transparently redirects them to the reorganized data. The SDS client library also provides a querying interface for reading part of the data based on user-specified selective criteria. We describe the design and implementation of the SDS client-server architecture, and evaluate the response time of the SDS Server and the performance benefits of SDS.« less

  16. Analyzing bean extracts using time-dependent SDS trapping to quantify the kinetic stability of phaseolin proteins.

    PubMed

    Thibeault, Jane; Church, Jennifer; Ortiz-Perez, Brian; Addo, Samuel; Hill, Shakeema; Khalil, Areeg; Young, Malaney; Xia, Ke; Colón, Wilfredo

    2017-09-30

    In common beans and lima bean, the storage protein phaseolin is difficult to degrade and SDS-resistant, a sign of kinetic stability. Kinetically stable proteins (KSPs) are characterized by having a high-energy barrier between the native and denatured states that results in very slow unfolding. Such proteins are resistant to proteolytic degradation and detergents, such as SDS. Here the method SDS-Trapping of Proteins (S-TraP) is applied directly on bean extracts to quantify the kinetic stability of phaseolin in lima bean and several common beans, including black bean, navy bean, and small red bean. The bean extracts were incubated in SDS at various temperatures (60-75 °C) for different time periods, followed by SDS-PAGE analysis at room temperature, and subsequent band quantification to determine the kinetics of phaseolin unfolding. Eyring plot analysis showed that the phaseolin from each bean has high kinetic stability, with an SDS-trapping (i.e. unfolding) half-life ranging from about 20-100 years at 24 °C and 2-7 years at 37 °C. The remarkably high kinetic stability of these phaseolin proteins is consistent with the low digestibility of common beans and lima bean, as well as their relatively high germination temperatures. From a practical perspective, this work exemplifies that S-TraP is a useful and cost-effective method for quantifying the kinetic stability of proteins in biological extracts or lysates. Depending on the protein to be studied and its abundance, S-TraP may be performed directly on the extract without need for protein purification. Copyright © 2017 Elsevier Inc. All rights reserved.

  17. How Old Is the New SDS?

    ERIC Educational Resources Information Center

    Isserman, Maurice

    2007-01-01

    Students for a Democratic Society (SDS) was the principal campus radical organization of the 1960s. When SDS first took form in 1960-62 under the leadership of Al Haber and Tom Hayden, it was a small organization of a few hundred members. By the time the author joined the Reed College chapter as a freshman in 1968, SDS had grown into a very large…

  18. Refolding of SDS-Unfolded Proteins by Nonionic Surfactants.

    PubMed

    Kaspersen, Jørn Døvling; Søndergaard, Anne; Madsen, Daniel Jhaf; Otzen, Daniel E; Pedersen, Jan Skov

    2017-04-25

    The strong and usually denaturing interaction between anionic surfactants (AS) and proteins/enzymes has both benefits and drawbacks: for example, it is put to good use in electrophoretic mass determinations but limits enzyme efficiency in detergent formulations. Therefore, studies of the interactions between proteins and AS as well as nonionic surfactants (NIS) are of both basic and applied relevance. The AS sodium dodecyl sulfate (SDS) denatures and unfolds globular proteins under most conditions. In contrast, NIS such as octaethylene glycol monododecyl ether (C 12 E 8 ) and dodecyl maltoside (DDM) protect bovine serum albumin (BSA) from unfolding in SDS. Membrane proteins denatured in SDS can also be refolded by addition of NIS. Here, we investigate whether globular proteins unfolded by SDS can be refolded upon addition of C 12 E 8 and DDM. Four proteins, BSA, α-lactalbumin (αLA), lysozyme, and β-lactoglobulin (βLG), were studied by small-angle x-ray scattering and both near- and far-UV circular dichroism. All proteins and their complexes with SDS were attempted to be refolded by the addition of C 12 E 8 , while DDM was additionally added to SDS-denatured αLA and βLG. Except for αLA, the proteins did not interact with NIS alone. For all proteins, the addition of NIS to the protein-SDS samples resulted in extraction of the SDS from the protein-SDS complexes and refolding of βLG, BSA, and lysozyme, while αLA changed to its NIS-bound state instead of the native state. We conclude that NIS competes with globular proteins for association with SDS, making it possible to release and refold SDS-denatured proteins by adding sufficient amounts of NIS, unless the protein also interacts with NIS alone. Copyright © 2017 Biophysical Society. Published by Elsevier Inc. All rights reserved.

  19. Development of the Space Debris Sensor (SDS)

    NASA Technical Reports Server (NTRS)

    Hamilton, J.; Liou, J.-C.; Anz-Meador, P. D.; Corsaro, B.; Giovane, F.; Matney, M.; Christiansen, E.

    2017-01-01

    The Space Debris Sensor (SDS) is a NASA experiment scheduled to fly aboard the International Space Station (ISS) starting in 2018. The SDS is the first flight demonstration of the Debris Resistive/Acoustic Grid Orbital NASA-Navy Sensor (DRAGONS) developed and matured at NASA Johnson Space Center's Orbital Debris Program Office. The DRAGONS concept combines several technologies to characterize the size, speed, direction, and density of small impacting objects. With a minimum two-year operational lifetime, SDS is anticipated to collect statistically significant information on orbital debris ranging from 50 microns to 500 microns in size. This paper describes the features of SDS and how data from the ISS mission may be used to update debris environment models. Results of hypervelocity impact testing during the development of SDS and the potential for improvement on future sensors at higher altitudes will be reviewed.

  20. Mass spectrometry-compatible silver staining of histones resolved on acetic acid-urea-Triton PAGE.

    PubMed

    Pramod, Khare Satyajeet; Bharat, Khade; Sanjay, Gupta

    2009-05-01

    Acetic acid-Urea-Triton (AUT) PAGE is commonly used method to separate histone variants and their post-translationally modified forms. Coomassie staining is the preferred method for protein visualization; however, its sensitivity is less than that of silver staining. Though silver staining of histones in AUT-PAGE has been reported, the method is time-consuming, dependent on prior staining by Amido black and has not been reported suitable for mass spectrometry. Here, we propose 'SDS-Silver' method for rapid, sensitive and mass spectrometry-compatible staining of histones resolved on AUT-PAGE.

  1. Sexual dimorphism in stature (SDS), jealousy and mate retention.

    PubMed

    Brewer, Gayle; Riley, Charlene

    2010-10-02

    Previous research has investigated the manner in which absolute height impacts on jealousy and mate retention. Although relative height is also important, little information exists about the potential influence of sexual dimorphism in stature (SDS) within established relationships. The current study investigated the relationship between SDS and the satisfaction, jealousy and mate retention behaviors reported by men and women. Heterosexual men (n = 98) and women (n = 102) completed a questionnaire. Men in high SDS relationships reported the lowest levels of cognitive and behavioral jealousy, although the impact of SDS on relationship satisfaction was less clear. SDS was not associated with the overall use of mate retention strategies; SDS did however affect the use of three specific strategies (vigilance, monopolization of time, love and care). SDS did not affect women's relationship satisfaction, jealousy (cognitive, behavioral, or emotional) or the use of mate retention strategies (with the exception of resource display).

  2. Isolation of thylakoid membrane complexes from rice by a new double-strips BN/SDS-PAGE and bioinformatics prediction of stromal ridge subunits interaction.

    PubMed

    Shao, Jinzhen; Zhang, Yubo; Yu, Jianlan; Guo, Lin; Ding, Yi

    2011-01-01

    Thylakoid membrane complexes of rice (Oryza sativa L.) play crucial roles in growth and crop production. Understanding of protein interactions within the complex would provide new insights into photosynthesis. Here, a new "Double-Strips BN/SDS-PAGE" method was employed to separate thylakoid membrane complexes in order to increase the protein abundance on 2D-gels and to facilitate the identification of hydrophobic transmembrane proteins. A total of 58 protein spots could be observed and subunit constitution of these complexes exhibited on 2D-gels. The generality of this new approach was confirmed using thylakoid membrane from spinach (Spinacia oleracea) and pumpkin (Cucurita spp). Furthermore, the proteins separated from rice thylakoid membrane were identified by the mass spectrometry (MS). The stromal ridge proteins PsaD and PsaE were identified both in the holo- and core- PSI complexes of rice. Using molecular dynamics simulation to explore the recognition mechanism of these subunits, we showed that salt bridge interactions between residues R19 of PsaC and E168 of PasD as well as R75 of PsaC and E91 of PsaD played important roles in the stability of the complex. This stromal ridge subunits interaction was also supported by the subsequent analysis of the binding free energy, the intramolecular distances and the intramolecular energy.

  3. Analysis of drought-tolerant sugar beet (Beta vulgaris L.) mutants induced with gamma radiation using SDS-PAGE and ISSR markers.

    PubMed

    Sen, Ayse; Alikamanoglu, Sema

    2012-01-01

    Drought is one of the major environmental stresses which greatly affect the plant growth and productivity. In the present study, various doses (0-75Gy) of gamma rays were applied to investigate the effect of radiation on shoot tip explants. It was observed that the regeneration rates and plant fresh weights decreased significantly with an increase in radiation dose. The optimal irradiation doses for mutation induction were determined at 15 and 20Gy. Afterwards, the induction of somatic mutation in sugar beet (Beta vulgaris L.) was investigated by irradiation of shoot tips with 15 and 20Gy gamma rays. Irradiated shoot tips were sub-cultured and M(1)V(1)-M(1)V(3) generations were obtained. Mutants tolerant to drought stress were selected on MS medium, supplemented with 10 and 20gl(-1) PEG6000. Of the M(1)V(3) plantlets, drought-tolerant mutants were selected. Leaf soluble proteins obtained from the control and drought-tolerant mutants were analyzed by SDS-PAGE. A total of 22 protein bands were determined and 2 of them were observed to be drought-tolerant mutants except the control. Polymorphism was also detected among the control and drought-tolerant mutants by DNA fingerprinting using ISSR markers. A total of 106 PCR fragments were amplified with 19 ISSR primers and 91 of them were polymorphic. The dendrograms were separated into two main clusters. First cluster included M8 mutant plant, which was applied 20Gy gamma radiation and regenerated on selective culture media containing 10gl(-1) PEG6000 concentration, and the second cluster was further divided into five sub-clusters. Copyright © 2012 Elsevier B.V. All rights reserved.

  4. Evaluation and standardization of different purification procedures for fish bile and liver metallothionein quantification by spectrophotometry and SDS-PAGE analyses.

    PubMed

    Tenório-Daussat, Carolina Lyrio; Resende, Marcia Carolina Martinho; Ziolli, Roberta L; Hauser-Davis, Rachel Ann; Schaumloffel, Dirk; Saint'Pierre, Tatiana D

    2014-03-01

    Fish bile metallothioneins (MT) have been recently reported as biomarkers for environmental metal contamination; however, no studies regarding standardizations for their purification are available. Therefore, different procedures (varying centrifugation times and heat-treatment temperatures) and reducing agents (DTT, β-mercaptoethanol and TCEP) were applied to purify MT isolated from fish (Oreochromis niloticus) bile and liver. Liver was also analyzed, since these two organs are intrinsically connected and show the same trend regarding MT expression. Spectrophotometrical analyses were used to quantify the resulting MT samples, and SDS-PAGE gels were used to qualitatively assess the different procedure results. Each procedure was then statistically evaluated and a multivariate statistical analysis was then applied. A response surface methodology was also applied for bile samples, in order to further evaluate the responses for this matrix. Heat treatment effectively removes most undesired proteins from the samples, however results indicate that temperatures above 70 °C are not efficient since they also remove MTs from both bile and liver samples. Our results also indicate that the centrifugation times described in the literature can be decreased in order to analyze more samples in the same timeframe, of importance in environmental monitoring contexts where samples are usually numerous. In an environmental context, biliary MT was lower than liver MT, as expected, since liver accumulates MT with slower detoxification rates than bile, which is released from the gallbladder during feeding, and then diluted by water. Therefore, bile MT seems to be more adequate in environmental monitoring scopes regarding recent exposure to xenobiotics that may affect the proteomic and metalloproteomic expression of this biological matrix. Copyright © 2013 Elsevier B.V. All rights reserved.

  5. Efficient renaturation of inclusion body proteins denatured by SDS.

    PubMed

    He, Chuan; Ohnishi, Kouhei

    2017-09-02

    Inclusion bodies are often formed when the foreign protein is over expressed in Escherichia coli. Since proteins in inclusion bodies are inactive, denaturing and refolding of inclusion body proteins are necessary to obtain the active form. Instead of the conventional denaturants, urea and guanidine hydrochloride, a strong anionic detergent SDS was used to solubilize C-terminal His-tag form of ulvan lyase in the inclusion bodies. Solution containing SDS-solubilized enzyme were kept on ice to precipitate SDS, followed by SDS-KCl insoluble crystal formation to remove SDS completely. After removing the precipitate by centrifugation, the supernatant was applied to Ni-NTA column to purify His-tagged ulvan lyase. The purified protein showed a dimeric form and ulvan lyase activity, demonstrating that SDS-denatured protein was renatured and recovered enzyme activity. This simple method could be useful for refolding other inclusion body proteins. Copyright © 2017 Elsevier Inc. All rights reserved.

  6. Two dimensional Blue Native-/SDS-PAGE analysis of SLP family adaptor protein complexes.

    PubMed

    Swamy, Mahima; Kulathu, Yogesh; Ernst, Sandra; Reth, Michael; Schamel, Wolfgang W A

    2006-04-15

    SH2 domain containing leukocyte protein (SLP) adaptor proteins serve a central role in the antigen-mediated activation of lymphocytes by organizing multiprotein signaling complexes. Here, we use two dimensional native-/SDS-gel electrophoresis to study the number, size and relative abundance of protein complexes containing SLP family proteins. In non-stimulated T cells all SLP-76 proteins are in a approximately 400 kDa complex with the small adaptor protein Grb2-like adaptor protein downstream of Shc (Gads), whereas half of Gads is monomeric. This constitutive SLP-76/Gads complex could be reconstituted in Drosophila S2 cells expressing both components, suggesting that it might not contain additional subunits. In contrast, in B cells SLP-65 exists in a 180 kDa complex as well as in monomeric form. Since the complex was not found in S2 cells expressing only SLP-65, it was not di/trimeric SLP-65. Upon antigen-stimulation only the complexed SLP-65 was phosphorylated. Surprisingly, stimulation-induced alteration of SLP complexes could not be detected, suggesting that active signaling complexes form only transiently, and are of low abundance.

  7. BMI and BMI SDS in childhood: annual increments and conditional change.

    PubMed

    Brannsether, Bente; Eide, Geir Egil; Roelants, Mathieu; Bjerknes, Robert; Júlíusson, Pétur Benedikt

    2017-02-01

    Background Early detection of abnormal weight gain in childhood may be important for preventive purposes. It is still debated which annual changes in BMI should warrant attention. Aim To analyse 1-year increments of Body Mass Index (BMI) and standardised BMI (BMI SDS) in childhood and explore conditional change in BMI SDS as an alternative method to evaluate 1-year changes in BMI. Subjects and methods The distributions of 1-year increments of BMI (kg/m 2 ) and BMI SDS are summarised by percentiles. Differences according to sex, age, height, weight, initial BMI and weight status on the BMI and BMI SDS increments were assessed with multiple linear regression. Conditional change in BMI SDS was based on the correlation between annual BMI measurements converted to SDS. Results BMI increments depended significantly on sex, height, weight and initial BMI. Changes in BMI SDS depended significantly only on the initial BMI SDS. The distribution of conditional change in BMI SDS using a two-correlation model was close to normal (mean = 0.11, SD = 1.02, n = 1167), with 3.2% (2.3-4.4%) of the observations below -2 SD and 2.8% (2.0-4.0%) above +2 SD. Conclusion Conditional change in BMI SDS can be used to detect unexpected large changes in BMI SDS. Although this method requires the use of a computer, it may be clinically useful to detect aberrant weight development.

  8. HFIP Extraction Followed by 2D CTAB/SDS-PAGE Separation: A New Methodology for Protein Identification from Tissue Sections after MALDI Mass Spectrometry Profiling for Personalized Medicine Research

    PubMed Central

    Longuespée, Rémi; Tastet, Christophe; Desmons, Annie; Kerdraon, Olivier; Day, Robert

    2014-01-01

    Abstract Matrix-assisted laser desorption ionization mass spectrometry imaging (MALDI-MSI) and profiling technology have become the easiest methods for quickly accessing the protein composition of a tissue area. Unfortunately, the demand for the identification of these proteins remains unmet. To overcome this bottleneck, we combined several strategies to identify the proteins detected via MALDI profiling including on-tissue protein extraction using hexafluoroIsopropanol (1,1,1,3,3,3-hexafluoro-2-propanol, HFIP) coupled with two-dimensional cetyl trimethylammonium bromide/sodium dodecyl sulfate–polyacrylamide gel electrophoresis (2D CTAB/SDS-PAGE) for separation followed by trypsin digestion and MALDI-MS analyses for identification. This strategy was compared with an on-tissue bottom-up strategy that we previously developed. The data reflect the complementarity of the approaches. An increase in the number of specific proteins identified has been established. This approach demonstrates the potential of adapted extraction procedures and the combination of parallel identification approaches for personalized medicine applications. The anatomical context provides important insight into identifying biomarkers and may be considered a first step for tissue-based biomarker research, as well as the extemporaneous examination of biopsies during surgery. PMID:24841221

  9. Sds22 regulates aurora B activity and microtubule–kinetochore interactions at mitosis

    PubMed Central

    Posch, Markus; Khoudoli, Guennadi A.; Swift, Sam; King, Emma M.; DeLuca, Jennifer G.

    2010-01-01

    We have studied Sds22, a conserved regulator of protein phosphatase 1 (PP1) activity, and determined its role in modulating the activity of aurora B kinase and kinetochore–microtubule interactions. Sds22 is required for proper progression through mitosis and localization of PP1 to mitotic kinetochores. Depletion of Sds22 increases aurora B T-loop phosphorylation and the rate of recovery from monastrol arrest. Phospho–aurora B accumulates at kinetochores in Sds22-depleted cells juxtaposed to critical kinetochore substrates. Sds22 modulates sister kinetochore distance and the interaction between Hec1 and the microtubule lattice and, thus, the activation of the spindle assembly checkpoint. These results demonstrate that Sds22 specifically defines PP1 function and localization in mitosis. Sds22 regulates PP1 targeting to the kinetochore, accumulation of phospho–aurora B, and force generation at the kinetochore–microtubule interface. PMID:20921135

  10. Best kept secrets ... Source Data Systems, Inc. (SDS).

    PubMed

    Andrew, W F

    1991-03-01

    The SDS/MEDNET system is a cost-effective option for small- to medium-size hospitals (up to 400 beds). The parameter-driven system lets users control operations with only occasional SDS assistance. A full application set, available for modular selection to reduce upfront costs while facilitating steady growth and protecting client investment, is adaptable to multi-facility environments. The industry-standard, Intel-based multi-user processors, network communications and protocols assure high efficiency, low-cost solutions independent of any one hardware vendor. Sustained growth in both client base and product offerings point to a high level of responsiveness and healthcare industry commitment. Corporate emphasis on user involvement and open systems integration assures clients of leading-edge capabilities. SDS/MEDNET will be a strong contender in selected marketing environments.

  11. Tunable smart digital structure (SDS) to modularly assemble soft actuators with layered adhesive bonding

    NASA Astrophysics Data System (ADS)

    Jin, Hu; Dong, Erbao; Xu, Min; Xia, Qirong; Liu, Shuai; Li, Weihua; Yang, Jie

    2018-01-01

    Many shape memory alloy (SMA)-based soft actuators have specific composite structures and manufacture processes, and are therefore unique. However, these exclusive characteristics limit their capabilities and applications, so in this article a soft and smart digital structure (SDS) is proposed that acts like a modular unit to assemble soft actuators by a layered adhesive bonding process. The SDS is a fully soft structure that encapsulates a digital skeleton consisting of four groups of parallel and independently actuated SMA wires capable of outputting a four-channel tunable force. The layered adhesive bonding process modularly bonds several SDSs with an elastic backbone to fabricate a layered soft actuator where the elastic backbone is used to recover the SDSs in a cooling process using the SMA wires. Two kinds of SDS-based soft actuators were modularly assembled, an actuator, SDS-I, with a two-dimensional reciprocal motion, and an actuator, SDS-II, capable of bi-directional reciprocal motion. The thermodynamics and phase transformation modeling of the SDS-based actuator were analyzed. Several extensional soft actuators were also assembled by bonding the SDS with an anomalous elastic backbone or modularly assembling the SDS-Is and SDS-IIs. These modularly assembled soft actuators delivered more output channels and a complicated motion, e.g., an actinomorphic soft actuator with four SDS-Is jumps in a series of hierarchical heights and directional movement by tuning the input channels of the SDSs. This result showed that the SDS can modularly assemble multifarious soft actuators with diverse capabilities, steerability and tunable outputs.

  12. Molecular dynamics simulation of sodium dodecylsulfate (SDS) bilayers.

    PubMed

    Zhang, Hongshu; Yuan, Shiling; Sun, Jichao; Liu, Jianqiang; Li, Haiping; Du, Na; Hou, Wanguo

    2017-11-15

    Sodium dodecylsulfate (SDS) - a simple single tailed surfactant (STS) can form stable vesicles from its micellar solution without any additives under the mediation of solid surfaces. To further understand the mechanism of this transition on the molecular level, molecular dynamics simulations are performed to study segments of SDS bilayers (as part of vesicles) in the bulk solution systematically, at the moment that the lower leaflet of bilayers already detached from solid surfaces. The SDS membrane would rather keep their bilayers structure than return to micelles when the initial interdigitated degree (δ i ) between alkyl chains is more than 8.0±1.4%. And the interdigitated degree is always approaching to 31.7±2.0% while the equilibrium is reached. The aggregates behave as curved bilayers, planar bilayers, perforated bilayers, and micelles with the increase of the lower leaflet cross-sectional area. Besides, the structures of salt bridge and water bridge structures are formed between DS - and Na + ions or water molecules, which contribute to the stability of SDS bilayers. The distribution difference of the salt bridges along the direction of S-O axis between the two leaflets leads to the asymmetry of the bilayers, which plays supplementary role to the formation of bilayers curvature. We expect that this work help to shed light on the understanding of interface phenomena and the mechanism of simple single-tailed surfactant vesicle self-assembly on the molecular level. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Sarcoidosis diagnostic score (SDS): a systematic evaluation to enhance the diagnosis of sarcoidosis.

    PubMed

    Bickett, Alexandra N; Lower, Elyse E; Baughman, Robert P

    2018-05-17

    The diagnosis of sarcoidosis is made by the combination of clinical features and biopsy results. The clinical features of sarcoidosis can be quite variable. We developed a Sarcoidosis Diagnostic Score (SDS) to summarize the clinical features of possible sarcoidosis patients. Biopsy confirmed sarcoidosis patients seen during a seven-month time period at the University of Cincinnati Sarcoidosis clinic were prospectively identified. Non-sarcoidosis patients seen at the same clinic were used as controls. Using a modified WASOG organ assessment instrument, we scored all patients for presence of biopsy, one or more highly probable symptom, and one or more at least probable symptom for each area. Two sarcoidosis scores were generated: SDS biopsy (with biopsy) and SDS clinical (without biopsy). The 980 evaluable patients were divided into two cohorts: an initial 600 patients (450 biopsy confirmed sarcoidosis, 150 controls) to establish cut-off values for SDS biopsy and SDS clinical and a validation cohort of 380 patients (103 biopsy confirmed sarcoidosis patients and 277 controls). The best cutoff value for SDS biopsy was > 6 (sensitivity =99.3%; specificity=100%). For the total the 980 patients, an SDS clinical > 3 had a sensitivity of 94.2%, specificity of 88.8%, and a likelihood ratio of 7.9. An SDS clinical score > 4 had a lower sensitivity of (76.9%) but higher specificity (98.6%). For sarcoidosis, the presence of specific clinical features, especially multi-organ involvement, can enhance the diagnostic certainty. The SDS scoring system quantitated the clinical features consistent with sarcoidosis. Copyright © 2018. Published by Elsevier Inc.

  14. Evaluation of SDS depletion using an affinity spin column and IMS-MS detection

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hengel, Shawna M.; Floyd, Erica A.; Baker, Erin Shammel

    2012-11-01

    While the use of detergents is necessary for a variety of protein isolation preparation protocols, often prior to mass spectral (MS) analysis, they are not compatible with MS analysis due to ion suppression and adduct formation. This manuscript describes optimization of detergent removal, using commercially available SDS depletion spin columns containing an affinity resin, providing for both increased protein recovery and thorough SDS removal. Ion mobility spectrometry coupled with mass spectrometry (IMS-MS) allowed for a concurrent analysis of both analyte and detergent. In the case of both proteins and peptides, higher detergent concentrations than previously reported provided an increase ofmore » sample recovery; however there was a limit as SDS was detected by IMS-MS at higher levels of SDS indicating incomplete detergent depletion. The results also suggest optimal conditions for SDS removal are dependent on the sample concentration. Overall, this study provides a useful guide for proteomic studies where SDS is required for efficient sample preparation.« less

  15. Small total dose measurement system for SDS-1

    NASA Astrophysics Data System (ADS)

    Kimoto, Yugo; Satoh, Yohei; Tachihara, Hiroshi

    2009-11-01

    The Japanese Aerospace Exploration Agency (JAXA) uses monitors on board satellites to measure and record in-flight data on ionization effects in space. A compact, total dose measurement system for the small satellite (SDS-1) was developed based on the previous system for measuring total ionizing dose effects. Especially, the sensor for SDS-1 is quite smaller than the sensor for SOHLA-1, which is presented in the last year. The sensor is 8 mm wide×3 mm high×19 mm long and weighs approximately 4 g with 500 mm its wire harness. Eight pin LCC RADFET and temperature sensor are arranged on it. Seven sensors are arranged on some components inside the SDS-1. One of the sensors is arranged on a printed board in advanced microprocessing in-ORBIT experiment equipment (AMI). The AMI demonstrate 320 MIPS microprocessor and DC-DC converter for space. The absorbed dose at the points where the sensors are arranged was evaluated before flight and will be compared with resulting flight data.

  16. Beyond the detergent effect: a binding site for sodium dodecyl sulfate (SDS) in mammalian apoferritin

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Liu, Renyu, E-mail: renyu.liu@uphs.upenn.edu; Bu, Weiming; Xi, Jin

    2012-05-01

    Using X-ray crystallography and isothermal titration calorimetry, we show that sodium dodecyl sulfate (SDS) binds specifically to a pre-formed internal cavity in horse-spleen apoferritin. Although sodium dodecyl sulfate (SDS) is widely used as an anionic detergent, it can also exert specific pharmacological effects that are independent of the surfactant properties of the molecule. However, structural details of how proteins recognize SDS are scarce. Here, it is demonstrated that SDS binds specifically to a naturally occurring four-helix bundle protein: horse apoferritin. The X-ray crystal structure of the apoferritin–SDS complex was determined at a resolution of 1.9 Å and revealed that themore » SDS binds in an internal cavity that has previously been shown to recognize various general anesthetics. A dissociation constant of 24 ± 9 µM at 293 K was determined by isothermal titration calorimetry. SDS binds in this cavity by bending its alkyl tail into a horseshoe shape; the charged SDS head group lies in the opening of the cavity at the protein surface. This crystal structure provides insights into the protein–SDS interactions that give rise to binding and may prove useful in the design of novel SDS-like ligands for some proteins.« less

  17. Interactions of PAMAM dendrimers with SDS at the solid-liquid interface.

    PubMed

    Arteta, Marianna Yanez; Eltes, Felix; Campbell, Richard A; Nylander, Tommy

    2013-05-14

    This work addresses structural and nonequilibrium effects of the interactions between well-defined cationic poly(amidoamine) PAMAM dendrimers of generations 4 and 8 and the anionic surfactant sodium dodecyl sulfate (SDS) at the hydrophilic silica-water interface. Neutron reflectometry and quartz crystal microbalance with dissipation monitoring were used to reveal the adsorption from premixed dendrimer/surfactant solutions as well as sequential addition of the surfactant to preadsorbed layers of dendrimers. PAMAM dendrimers of both generations adsorb to hydrophilic silica as a compact monolayer, and the adsorption is irreversible upon rinsing with salt solution. SDS adsorbs on the dendrimer layer and at low bulk concentrations causes the expansion of the dendrimer layers on the surface. When the bulk concentration of SDS is increased, the surfactant layer consists of aggregates or bilayer-like structures. The adsorption of surfactant is reversible upon rinsing, but slight changes of the structure of the preadsorbed PAMAM monolayer were observed. The adsorption from premixed solutions close to charge neutrality results in thick multilayers, but the surface excess is lower when the bulk complexes have a net negative charge. A critical examination of the pathway of adsorption for the interactions of SDS with preadsorbed PAMAM monolayers and premixed PAMAM/SDS solutions with hydrophilic silica revealed that nonequilibrium effects are important only in the latter case, and the application of a thermodynamic model to such experimental data would be inappropriate.

  18. Gallium(III)/4-(2-pyridylazo)resorcinol system in water and SDS solution: kinetics and thermodynamics.

    PubMed

    Biver, T; Boggioni, A; Secco, F; Venturini, M

    2008-01-01

    The equilibria and kinetics of the complex formation and dissociation reaction between gallium(III) and PAR [4-(2-pyridylazo)resorcinol] have been investigated in water and in the presence of SDS micelles. The reactive form of Ga(III) is GaOH2+ in both cases. The addition of SDS results in an increase of both the binding affinity and velocity, the maximum accelerating effect being observed just above the cmc value of SDS that, under the conditions of the experiments, is 5.6 x 10-3 M. At pH = 3.2, the maximum value of the equilibrium constant ratio Kapp(SDS)/Kapp(H2O) is 27.4, whereas that of the binding rate constants kf(SDS)/kf(H2O) is 16. The results are interpreted in terms of increased concentrations of the reactants on the micelle surface and on competition of PAR and SDS for GaOH2+.

  19. Phosphorylation of PP1 Regulator Sds22 by PLK1 Ensures Accurate Chromosome Segregation.

    PubMed

    Duan, Hequan; Wang, Chunli; Wang, Ming; Gao, Xinjiao; Yan, Maomao; Akram, Saima; Peng, Wei; Zou, Hanfa; Wang, Dong; Zhou, Jiajia; Chu, Youjun; Dou, Zhen; Barrett, Gregory; Green, Hadiyah-Nichole; Wang, Fangjun; Tian, Ruijun; He, Ping; Wang, Wenwen; Liu, Xing; Yao, Xuebiao

    2016-09-30

    During cell division, accurate chromosome segregation is tightly regulated by Polo-like kinase 1 (PLK1) and opposing activities of Aurora B kinase and protein phosphatase 1 (PP1). However, the regulatory mechanisms underlying the aforementioned hierarchical signaling cascade during mitotic chromosome segregation have remained elusive. Sds22 is a conserved regulator of PP1 activity, but how it regulates PP1 activity in space and time during mitosis remains elusive. Here we show that Sds22 is a novel and cognate substrate of PLK1 in mitosis, and the phosphorylation of Sds22 by PLK1 elicited an inhibition of PP1-mediated dephosphorylation of Aurora B at threonine 232 (Thr 232 ) in a dose-dependent manner. Overexpression of a phosphomimetic mutant of Sds22 causes a dramatic increase in mitotic delay, whereas overexpression of a non-phosphorylatable mutant of Sds22 results in mitotic arrest. Mechanistically, the phosphorylation of Sds22 by PLK1 strengthens the binding of Sds22 to PP1 and inhibits the dephosphorylation of Thr 232 of Aurora B to ensure a robust, error-free metaphase-anaphase transition. These findings delineate a conserved signaling hierarchy that orchestrates dynamic protein phosphorylation and dephosphorylation of critical mitotic regulators during chromosome segregation to guard chromosome stability. © 2016 by The American Society for Biochemistry and Molecular Biology, Inc.

  20. Small Total Dose Measurement System for SOHLA-1 and SDS-1

    NASA Astrophysics Data System (ADS)

    Kimoto, Yugo; Satoh, Yohei; Tachihara, Hiroshi

    The Japanese Aerospace Exploration Agency (JAXA) uses monitors on board satellites to measure and record in-flight data about ionization effects in space. A compact, total-dose measurement system for small satellites—Space-Oriented Higashiosaka Leading Association -1 (SOHLA-1) and Small Demonstration-Satellite -1 (SDS-1)—was developed based on a prior system for measuring total ionizing dose effects. Especially, the sensor for SDS-1 is much smaller than the sensor for SOHLA-1. The sensor for SDS-1 is 8 mm wide × 3 mm high × 19 mm long and weighs approximately 4 g with 500 mm with its wire harness. An 8-pin Lead less Chip Carrier (LCC) RADFET and temperature sensor are arranged on it. Seven sensors are mounted on some components inside the SDS-1. The sensor for SOHLA-1 is a 14-pin Dual Inline Package (DIP) type RADFET. The four sensors, which have RADFET on a printed board covered with an aluminum chassis, are mounted both inside and outside the satellite. This report presents small total dose measurement systems and ground irradiation test results for two small satellites.

  1. NMR investigations of self-aggregation characteristics of SDS in a model assembled tri-block copolymer solution.

    PubMed

    Kumar, B V N Phani; Priyadharsini, S Umayal; Prameela, G K S; Mandal, Asit Baran

    2011-08-01

    The present work was undertaken with a view to understand the influence of a model non-ionic tri-block copolymer PEO-PPO-PEO (poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide)) with molecular weight 5800 i.e., P123 [(EO)(20)-(PO)(70)-(EO)(20)] on the self-aggregation characteristics of the anionic surfactant sodium dodecylsulfate (SDS) in aqueous solution (D(2)O) using NMR chemical shift, self-diffusion and nuclear spin-relaxation as suitable experimental probes. In addition, polymer diffusion has been monitored as a function of SDS concentration. The concentration-dependent chemical shift, diffusion data and relaxation data indicated the significant interaction of polymeric micelles with SDS monomers and micelles at lower and intermediate concentrations of SDS, whereas the weak interaction of the polymer with SDS micelles at higher concentrations of SDS. It has been observed that SDS starts aggregating on the polymer at a lower concentration i.e., critical aggregation concentration (cac=1.94 mM) compared to polymer-free situation, and the onset of secondary micelle concentration (C(2)=27.16 mM) points out the saturation of the 0.2 wt% polymer or free SDS monomers/micelles at higher concentrations of SDS. It has also been observed that the parameter cac is almost independent in the polymer concentrations of study. The TMS (tetramethylsilane) has been used as a solubilizate to measure the bound diffusion coefficient of SDS-polymer mixed system. The self-diffusion data were analyzed using two-site exchange model and the obtained information on aggregation dynamics was commensurate with that inferred from chemical shift and relaxation data. The information on slow motions of polymer-SDS system was also extracted using spin-spin and spin-lattice relaxation rate measurements. The relaxation data points out the disintegration of polymer network at higher concentrations of SDS. The present NMR investigations have been well corroborated by surface tension and

  2. Childhood obesity treatment; Effects on BMI SDS, body composition, and fasting plasma lipid concentrations.

    PubMed

    Nielsen, Tenna Ruest Haarmark; Fonvig, Cilius Esmann; Dahl, Maria; Mollerup, Pernille Maria; Lausten-Thomsen, Ulrik; Pedersen, Oluf; Hansen, Torben; Holm, Jens-Christian

    2018-01-01

    The body mass index (BMI) standard deviation score (SDS) may not adequately reflect changes in fat mass during childhood obesity treatment. This study aimed to investigate associations between BMI SDS, body composition, and fasting plasma lipid concentrations at baseline and during childhood obesity treatment. 876 children and adolescents (498 girls) with overweight/obesity, median age 11.2 years (range 1.6-21.7), and median BMI SDS 2.8 (range 1.3-5.7) were enrolled in a multidisciplinary outpatient treatment program and followed for a median of 1.8 years (range 0.4-7.4). Height and weight, body composition measured by dual-energy X-ray absorptiometry, and fasting plasma lipid concentrations were assessed at baseline and at follow-up. Lipid concentrations (total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), non-HDL, and triglycerides (TG)) were available in 469 individuals (264 girls). Linear regressions were performed to investigate the associations between BMI SDS, body composition indices, and lipid concentrations. At baseline, BMI SDS was negatively associated with concentrations of HDL (p = 6.7*10-4) and positively with TG (p = 9.7*10-6). Reductions in BMI SDS were associated with reductions in total body fat percentage (p<2*10-16) and percent truncal body fat (p<2*10-16). Furthermore, reductions in BMI SDS were associated with improvements in concentrations of TC, LDL, HDL, non-HDL, LDL/HDL-ratio, and TG (all p <0.0001). Changes in body fat percentage seemed to mediate the changes in plasma concentrations of TC, LDL, and non-HDL, but could not alone explain the changes in HDL, LDL/HDL-ratio or TG. Among 81 individuals with available lipid concentrations, who increased their BMI SDS, 61% improved their body composition, and 80% improved their lipid concentrations. Reductions in the degree of obesity during multidisciplinary childhood obesity treatment are accompanied by improvements in body composition and fasting plasma

  3. Nutritional status in sick children and adolescents is not accurately reflected by BMI-SDS.

    PubMed

    Fusch, Gerhard; Raja, Preeya; Dung, Nguyen Quang; Karaolis-Danckert, Nadina; Barr, Ronald; Fusch, Christoph

    2013-01-01

    Nutritional status provides helpful information of disease severity and treatment effectiveness. Body mass index standard deviation scores (BMI-SDS) provide an approximation of body composition and thus are frequently used to classify nutritional status of sick children and adolescents. However, the accuracy of estimating body composition in this population using BMI-SDS has not been assessed. Thus, this study aims to evaluate the accuracy of nutritional status classification in sick infants and adolescents using BMI-SDS, upon comparison to classification using percentage body fat (%BF) reference charts. BMI-SDS was calculated from anthropometric measurements and %BF was measured using dual-energy x-ray absorptiometry (DXA) for 393 sick children and adolescents (5 months-18 years). Subjects were classified by nutritional status (underweight, normal weight, overweight, and obese), using 2 methods: (1) BMI-SDS, based on age- and gender-specific percentiles, and (2) %BF reference charts (standard). Linear regression and a correlation analysis were conducted to compare agreement between both methods of nutritional status classification. %BF reference value comparisons were also made between 3 independent sources based on German, Canadian, and American study populations. Correlation between nutritional status classification by BMI-SDS and %BF agreed moderately (r (2) = 0.75, 0.76 in boys and girls, respectively). The misclassification of nutritional status in sick children and adolescents using BMI-SDS was 27% when using German %BF references. Similar rates observed when using Canadian and American %BF references (24% and 23%, respectively). Using BMI-SDS to determine nutritional status in a sick population is not considered an appropriate clinical tool for identifying individual underweight or overweight children or adolescents. However, BMI-SDS may be appropriate for longitudinal measurements or for screening purposes in large field studies. When accurate nutritional

  4. Medium dependent dual turn on/turn off fluorescence sensing for Cu2 + ions using AMI/SDS assemblies

    NASA Astrophysics Data System (ADS)

    Gujar, Varsha B.; Ottoor, Divya

    2017-02-01

    Behavior of Amiloride (AMI) as a metal ion sensor in anionic surfactant assemblies of varying concentrations at different pH is depicted in this work. From a non-sensor fluorophore, AMI has been transformed in to a tunable fluorosensor for Cu2 + ions in various SDS concentrations. At premicellar concentration of SDS, ion-pair complex is expected to be formed between AMI and SDS due to electrostatic interactions between them. However at CMC concentrations of SDS, fluorescence intensity of AMI is greatly enhanced with red shift in emission, due to the incorporation of AMI molecule in the hydrophobic micellar interface. The behavior of metal sensing by AMI-SDS assemblies gives rise to several interesting observations. Micellation of SDS has been greatly enhanced by increasing copper ion concentrations, as these counter ions screens the charge on monomers of SDS which lead to the aggregation at premicellar concentrations only. Concentrations and pH dependent discrete trends of interactions between SDS-AMI and SDS-Cu2 + ions, have given tunable fluorescence responses (fluorescence turn on/turn off) of AMI for added Cu2 + ions. The electrostatic interaction between the metal cations and the anionic surfactants is the driving force for bringing the metal ions near to the vicinity of micelle where AMI resides. Thus, a comprehensive understanding of the mechanism related to the 'turn on-turn off' fluorescence response of AMI with respect to pH and SDS concentration for effective Cu2 + ion sensing is illustrated in this work.

  5. Childhood obesity treatment; Effects on BMI SDS, body composition, and fasting plasma lipid concentrations

    PubMed Central

    Fonvig, Cilius Esmann; Dahl, Maria; Mollerup, Pernille Maria; Lausten-Thomsen, Ulrik; Pedersen, Oluf; Hansen, Torben; Holm, Jens-Christian

    2018-01-01

    Objective The body mass index (BMI) standard deviation score (SDS) may not adequately reflect changes in fat mass during childhood obesity treatment. This study aimed to investigate associations between BMI SDS, body composition, and fasting plasma lipid concentrations at baseline and during childhood obesity treatment. Methods 876 children and adolescents (498 girls) with overweight/obesity, median age 11.2 years (range 1.6–21.7), and median BMI SDS 2.8 (range 1.3–5.7) were enrolled in a multidisciplinary outpatient treatment program and followed for a median of 1.8 years (range 0.4–7.4). Height and weight, body composition measured by dual-energy X-ray absorptiometry, and fasting plasma lipid concentrations were assessed at baseline and at follow-up. Lipid concentrations (total cholesterol (TC), low-density lipoprotein (LDL), high-density lipoprotein (HDL), non-HDL, and triglycerides (TG)) were available in 469 individuals (264 girls). Linear regressions were performed to investigate the associations between BMI SDS, body composition indices, and lipid concentrations. Results At baseline, BMI SDS was negatively associated with concentrations of HDL (p = 6.7*10−4) and positively with TG (p = 9.7*10−6). Reductions in BMI SDS were associated with reductions in total body fat percentage (p<2*10−16) and percent truncal body fat (p<2*10−16). Furthermore, reductions in BMI SDS were associated with improvements in concentrations of TC, LDL, HDL, non-HDL, LDL/HDL-ratio, and TG (all p <0.0001). Changes in body fat percentage seemed to mediate the changes in plasma concentrations of TC, LDL, and non-HDL, but could not alone explain the changes in HDL, LDL/HDL-ratio or TG. Among 81 individuals with available lipid concentrations, who increased their BMI SDS, 61% improved their body composition, and 80% improved their lipid concentrations. Conclusion Reductions in the degree of obesity during multidisciplinary childhood obesity treatment are accompanied by

  6. Screening of SDS-degrading bacteria from car wash wastewater and study of the alkylsulfatase enzyme activity

    PubMed Central

    Shahbazi, Razieh; Kasra-Kermanshahi, Roha; Gharavi, Sara; Moosavi-Nejad, Zahra; Borzooee, Faezeh

    2013-01-01

    Background and Objectives Sodium dodecyl sulfate (SDS) is one of the main surfactant components in detergents and cosmetics, used in high amounts as a detergent in products such as shampoos, car wash soap and toothpaste. Therefore, its bioremediation by suitable microorganisms is important. Alkylsulfatase is an enzyme that hydrolyses sulfate -ester bonds to give inorganic sulfate and alcohol. The purpose of this study was to isolate SDS–degrading bacteria from Tehran city car wash wastewater, study bacterial alkylsulfatase enzyme activity and identify the alkylsulfatase enzyme coding gene. Materials and Methods Screening of SDS-degrading bacteria was carried out on basal salt medium containing SDS as the sole source of carbon. Amount of SDS degraded was assayed by methylene blue active substance (MBAS). Results and Conclusion Identification of the sdsA gene was carried by PCR and subsequent sequencing of the 16S rDNA gene and biochemical tests identified Pseudomonas aeruginosa. This bacterium is able to degrade 84% of SDS after four days incubation. Bacteria isolated from car wash wastewater were shown to carry the sdsA gene (670bp) and the alkylsulfatase enzyme specific activity expressed from this gene was determined to be 24.3 unit/mg. The results presented in this research indicate that Pseudomonas aeruginosa is a suitable candidate for SDS biodegradation. PMID:23825734

  7. The predictive power of SIMION/SDS simulation software for modeling ion mobility spectrometry instruments

    NASA Astrophysics Data System (ADS)

    Lai, Hanh; McJunkin, Timothy R.; Miller, Carla J.; Scott, Jill R.; Almirall, José R.

    2008-09-01

    The combined use of SIMION 7.0 and the statistical diffusion simulation (SDS) user program in conjunction with SolidWorks® with COSMSOSFloWorks® fluid dynamics software to model a complete, commercial ion mobility spectrometer (IMS) was demonstrated for the first time and compared to experimental results for tests using compounds of immediate interest in the security industry (e.g., 2,4,6-trinitrotoluene, 2,7-dinitrofluorene, and cocaine). The effort of this research was to evaluate the predictive power of SIMION/SDS for application to IMS instruments. The simulation was evaluated against experimental results in three studies: (1) a drift:carrier gas flow rates study assesses the ability of SIMION/SDS to correctly predict the ion drift times; (2) a drift gas composition study evaluates the accuracy in predicting the resolution; (3) a gate width study compares the simulated peak shape and peak intensity with the experimental values. SIMION/SDS successfully predicted the correct drift time, intensity, and resolution trends for the operating parameters studied. Despite the need for estimations and assumptions in the construction of the simulated instrument, SIMION/SDS was able to predict the resolution between two ion species in air within 3% accuracy. The preliminary success of IMS simulations using SIMION/SDS software holds great promise for the design of future instruments with enhanced performance.

  8. The Predictive Power of SIMION/SDS Simulation Software for Modeling Ion Mobility Spectrometry Instruments

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hanh Lai; Timothy R. McJunkin; Carla J. Miller

    2008-09-01

    The combined use of SIMION 7.0 and the statistical diffusion simulation (SDS) user program in conjunction with SolidWorks® with COSMSOFloWorks® fluid dynamics software to model a complete, commercial ion mobility spectrometer (IMS) was demonstrated for the first time and compared to experimental results for tests using compounds of immediate interest in the security industry (e.g., 2,4,6-trinitrotoluene and cocaine). The effort of this research was to evaluate the predictive power of SIMION/SDS for application to IMS instruments. The simulation was evaluated against experimental results in three studies: 1) a drift:carrier gas flow rates study assesses the ability of SIMION/SDS to correctlymore » predict the ion drift times; 2) a drift gas composition study evaluates the accuracy in predicting the resolution; and 3) a gate width study compares the simulated peak shape and peak intensity with the experimental values. SIMION/SDS successfully predicted the correct drift time, intensity, and resolution trends for the operating parameters studied. Despite the need for estimations and assumptions in the construction of the simulated instrument, SIMION/SDS was able to predict the resolution between two ion species in air within 3% accuracy. The preliminary success of IMS simulations using SIMION/SDS software holds great promise for the design of future instruments with enhanced performance.« less

  9. Effect of SDS on human hair: Study on the molecular structure and morphology.

    PubMed

    Singh, Bhawana; Umapathy, Siva

    2011-05-01

    This paper presents a model study to understand the effect of surfactants on the physicochemical properties of human hair. FT-IR ATR spectroscopy has been employed to understand the chemical changes induced by sodium dodecyl sulfate (SDS) on human scalp hair. In particular, the SDS induced changes in the secondary structure of protein present in the outer protective layer of hair, i.e. cuticle, have been investigated. Conformational changes in the secondary structure of protein were studied by curve fitting of the amide I band after every phase of SDS treatment. It has been found that SDS brings rearrangements in the protein backbone conformations by transforming β -sheet structure to random coil and β -turn. Additionally, AFM and SEM studies were carried out to understand the morphological changes induced on the hair surface. SEM and AFM images demonstrated the rupture and partial erosion of cuticle sublayers. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  10. Identification and In Silico Analysis of Major Redox Modulated Proteins from Brassica juncea Seedlings Using 2D Redox SDS PAGE (2-Dimensional Diagonal Redox Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis).

    PubMed

    Chaurasia, Satya Prakash; Deswal, Renu

    2017-02-01

    The thiol-disulphide exchange regulates the activity of proteins by redox modulation. Many studies to analyze reactive oxygen species (ROS), particularly, hydrogen peroxide (H 2 O 2 ) induced changes in the gene expression have been reported, but efforts to detect H 2 O 2 modified proteins are comparatively few. Two-dimensional diagonal redox sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) was used to detect polypeptides which undergo thiol-disulphide exchange in Brassica juncea seedlings following H 2 O 2 (10 mM) treatment for 30 min. Eleven redox responsive polypeptides were identified which included cruciferin, NLI [Nuclear LIM (Lin11, Isl-1 & Mec-3 domains)] interacting protein phosphatase, RuBisCO (ribulose-1,5-bisphosphate carboxylase/oxygenase) large subunit, and myrosinase. Redox modulation of RuBisCO large subunit was further confirmed by western blotting. However, the small subunit of RuBisCO was not affected by these redox changes. All redox modulated targets except NLI interacting protein (although it contains two cysteines) showed oxidation sensitive cysteines by in silico analysis. Interestingly, interactome of cruciferin and myrosinase indicated that they may have additional function(s) beside their well-known roles in the seedling development and abiotic stress respectively. Cruciferin showed interactions with stress associated proteins like defensing-like protein 192 and 2-cys peroxiredoxin. Similarly, myrosinase showed interactions with nitrilase and cytochrome p450 which are involved in nitrogen metabolism and/or hormone biosynthesis. This simple procedure can be used to detect major stress mediated redox changes in other plants.

  11. Physicochemical perspectives (aggregation, structure and dynamics) of interaction between pluronic (L31) and surfactant (SDS).

    PubMed

    Prameela, G K S; Phani Kumar, B V N; Pan, A; Aswal, V K; Subramanian, J; Mandal, A B; Moulik, S P

    2015-11-11

    The influence of the water soluble non-ionic tri-block copolymer PEO-PPO-PEO [poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide)] i.e., E2P16E2 (L31) on the microstructure and self-aggregation dynamics of the anionic surfactant sodium dodecylsulfate (SDS) in aqueous solution was investigated using cloud point (CP), isothermal titration calorimetry (ITC), high resolution nuclear magnetic resonance (NMR), electron paramagnetic resonance (EPR), and small-angle neutron scattering (SANS) measurements. CP provided the thermodynamic information on the Gibbs free energy, enthalpy, entropy and heat capacity changes pertaining to the phase separation of the system at elevated temperature. The ITC and NMR self-diffusion measurements helped to understand the nature of the binding isotherms of SDS in the presence of L31 in terms of the formation of mixed aggregates and free SDS micelles in solution. EPR analysis provided the micro-viscosity of the spin probe 5-DSA in terms of rotational correlation time. The SANS study indicated the presence of prolate ellipsoidal mixed aggregates, whose size increased with the increasing addition of L31. At a large [L31], SANS also revealed the progressive decreasing size of the ellipsoidal mixed aggregates of SDS-L31 into nearly globular forms with the increasing SDS addition. Wrapping of the spherical SDS micelles by L31 was also corroborated from (13)C NMR and SANS measurements.

  12. [Schedule for evaluation of the deficit syndrome in schizophrenia: Schedule for Deficit Syndrome (SDS) (Kirkpatrick et al.). Importance pertinence of the SDS. Introduction of the French version].

    PubMed

    Ribeyre, J M; Dollfus, S; Lesieur, P; Ménard, J F; Petit, M

    1994-01-01

    The negative symptoms of schizophrenia have generated a great interest leading some authors (Crow, Andreasen, Kay) to delineate schizophrenic subtypes based on their presence or absence. Carpenter et al. have recently proposed another subtype, the deficit syndrome, based on Kraepelin's clinical description. This differs from other proposed negative subtypes and refers to the presence or absence of prominent, enduring and primary negative symptoms. Primary negative symptoms have to be due to psychophrenia itself, in other words, independent of factors such as depression, anxiety, akinesia... Kirkpatrick et al. have proposed the Schedule for the Deficit Syndrome (SDS) to reliably identify this deficit syndrome. Some studies using this instrument have supported the validity of the deficit syndrome concept. Particularly, deficit patients have clinical, neuropsychological, neurological, eye-tracking and brain imaging impairments compared to nondeficit patients. We realized a french translation of SDS and used it to study a biological index (plasma homovanillic acid, pHVA) among deficit and nondeficit schizophrenic patients. Our data suggest a specific biochemical basis for the deficit syndrome, ie, significant lower mean pHVA levels with a lack of diurnal variation for deficit patients. The french version of SDS was validated by Kirkpatrick after english back translation. We present here our psychometric data regarding reliability (assessed by weighted and unweighted kappa coefficients) and cohesiveness of the construct (assessed by rank-order correlations of each negative symptoms with the other five, using Spearman's rho). These data are quite significant and in agreement with the SDS authors.

  13. Highly thermostable, thermophilic, alkaline, SDS and chelator resistant amylase from a thermophilic Bacillus sp. isolate A3-15.

    PubMed

    Arikan, Burhan

    2008-05-01

    A thermostable alkaline alpha-amylase producing Bacillus sp. A3-15 was isolated from compost samples. There was a slight variation in amylase synthesis within the pH range 6.0 and 12.0 with an optimum pH of 8.5 (8mm zone diameter in agar medium) on starch agar medium. Analyses of the enzyme for molecular mass and amylolytic activity were carried out by starch SDS-PAGE electrophoresis, which revealed two independent bands (86,000 and 60,500 Da). Enzyme synthesis occurred at temperatures between 25 and 65 degrees C with an optimum of 60 degrees C on petri dishes. The partial purification enzyme showed optimum activity at pH 11.0 and 70 degrees C. The enzyme was highly active (95%) in alkaline range of pH (10.0-11.5), and it was almost completely active up to 100 degrees C with 96% of the original activity remaining after heat treatment at 100 degrees C for 30 min. Enzyme activity was enhanced in the presence of 5mM CaCl2 (130%) and inhibition with 5mM by ZnCl2, NaCl, Na-sulphide, EDTA, PMSF (3mM), Urea (8M) and SDS (1%) was obtained 18%, 20%, 36%, 5%, 10%, 80% and 18%, respectively. The enzyme was stable approximately 70% at pH 10.0-11.0 and 60 degrees C for 24h. So our result showed that the enzyme was both, highly thermostable-alkaline, thermophile and chelator resistant. The A3-15 amylase enzyme may be suitable in liquefaction of starch in high temperature, in detergent and textile industries and in other industrial applications.

  14. Isolation and identification of two novel SDS-resistant secreted chitinases from Aeromonas schubertii

    PubMed Central

    Liu, Chao-Lin; Shen, Chia-Rui; Hsu, Fong-Fu; Chen, Jeen-Kuan; Wu, Pei-Tzu; Guo, Shang-Hsin; Lee, Wen-Chien; Yu, Feng-Wei; Mackey, Zachary B.; Turk, John; Gross, Michael L.

    2008-01-01

    Two SDS-resistant endochitinases, designated as ASCHI53 and ASCHI61, were isolated from Aeromonas schubertii in a soil sample from southern Taiwan. MALDI-TOF mass measurement indicates the molecular weights of 53,527 for ASCHI53 and 61,202 for ASCHI61. N-terminal and internal amino acid sequences were obtained, and BLAST analysis of the sequences and MS/MS peptide sequencing showed that they were novel proteins. Degradation of chitin by these two endochitinases gave rise to hexameric chitin oligosaccharide, a compound known to have several potent biomedical functions. ASCHI53 and ASCHI61 retained, respectively, 65% and 75%, of their chitinase activity in the presence of 5% SDS and 100% of their activity in the presence of 10% β-mercaptoethanol. These results demonstrate that they are SDS-resistant endochitinases and probably have a rigid structure. PMID:19197977

  15. Microfluidic device having an immobilized pH gradient and PAGE gels for protein separation and analysis

    DOEpatents

    Sommer, Gregory J.; Hatch, Anson V.; Singh, Anup K.; Wang, Ying-Chih

    2012-12-11

    Disclosed is a novel microfluidic device enabling on-chip implementation of a two-dimensional separation methodology. Previously disclosed microscale immobilized pH gradients (IPG) are combined with perpendicular polyacrylamide gel electrophoresis (PAGE) microchannels to achieve orthogonal separations of biological samples. Device modifications enable inclusion of sodium dodecyl sulfate (SDS) in the second dimension. The device can be fabricated to use either continuous IPG gels, or the microscale isoelectric fractionation membranes we have also previously disclosed, for the first dimension. The invention represents the first all-gel two-dimensional separation microdevice, with significantly higher resolution power over existing devices.

  16. Microfluidic device having an immobilized pH gradient and page gels for protein separation and analysis

    DOEpatents

    Sommer, Gregory J; Hatch, Anson V; Singh, Anup K; Wang, Ying-Chih

    2014-05-20

    Disclosed is a novel microfluidic device enabling on-chip implementation of a two-dimensional separation methodology. Previously disclosed microscale immobilized pH gradients (IPG) are combined with perpendicular polyacrylamide gel electrophoresis (PAGE) microchannels to achieve orthogonal separations of biological samples. Device modifications enable inclusion of sodium dodecyl sulfate (SDS) in the second dimension. The device can be fabricated to use either continuous IPG gels, or the microscale isoelectric fractionation membranes we have also previously disclosed, for the first dimension. The invention represents the first all-gel two-dimensional separation microdevice, with significantly higher resolution power over existing devices.

  17. WMO Sand and Dust Storm Warning Advisory and Assessment System (SDS-WAS): Regional Cooperation Initiative for Americas

    NASA Astrophysics Data System (ADS)

    Nickovic, S.; Sprigg, W. A.

    2011-12-01

    For countries in and downwind of arid regions, airborne sand and dust present serious risks to the environment, property and human health. The World Meteorological Organization (WMO) has taken the lead on an international scale to develop and implement SDS-WAS - a system of dust-related research, observations, numerical dust prediction and services. Partnership in SDS-WAS is based on a federated 'open-club' principle. SDS-WAS realizes its goals by networking research institutions, operational centers and users organized through regional nodes assisted by associated regional centers. Growing interest and the needs of communities in the Americas call for establishing a Pan-American SDS-WAS node to serve as an information clearinghouse and to facilitate collaboration that will lead to greater economy of effort and speedier transition of research to applications.

  18. Erythrocyte membrane protein analysis by sodium dodecyl sulphate-capillary gel electrophoresis in the diagnosis of hereditary spherocytosis.

    PubMed

    Debaugnies, France; Cotton, Frédéric; Boutique, Charles; Gulbis, Béatrice

    2011-03-01

    Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) is currently the reference method for detecting protein deficiencies related to hereditary spherocytosis. The aim of the study was to evaluate an automated capillary gel electrophoresis system, the Experion instrument from BioRad, for its ability to separate and quantify the erythrocyte membrane proteins. The major erythrocyte membrane proteins (actin, protein 4.2, protein 4.1, band 3, ankyrin, α- and β-spectrin) were extracted and purified from membrane ghosts by centrifugation, immunoprecipitation and electroelution. Analyses were performed using SDS-PAGE and sodium dodecyl sulphate capillary gel electrophoresis (SDS-CGE) to establish a separation profile of the total ghosts. Then, the samples from patients received for investigations of erythrocyte membrane defects were analysed. Five of the seven expected erythrocyte membrane proteins were finally separated and identified. In the 20 studied cases, taking into account the screening test results and the clinical and family histories, the SDS-CGE method allowed us to achieve the same conclusion as with SDS-PAGE, except for the patient with elliptocytosis. The new SDS-CGE method presents interesting features that could make this instrument a powerful diagnostic tool for detection of erythrocyte membrane protein abnormalities, and can be proposed as an automated alternative method to the labour intensive SDS-PAGE analysis.

  19. Stress responses of duckweed (Lemna minor L.) and water velvet (Azolla filiculoides Lam.) to anionic surfactant sodium-dodecyl-sulphate (SDS).

    PubMed

    Forni, C; Braglia, R; Harren, F J M; Cristescu, S M

    2012-04-01

    Surfactants are used for several purposes and recently they have attracted the attention for their ability to modify the behavior of other preexistent or co-disposed contaminants, although their use or discharge in wastewaters can represent a real or potential risk for the environment. Lemna minor L. and Azolla filiculoides Lam. are floating aquatic macrophytes, very effective in accumulating several pollutants including sodium dodecyl sulphate (SDS). In this work we evaluated the effects of SDS on these species by determining the stress ethylene production via laser-based trace gas detection, and the activities of enzymes involved in stress response, such as guaiacol peroxidase (G-POD), phenylalanine ammonia-lyase (PAL) and polyphenol-oxidase (PPO). Phenolics content was also determined. The macrophytes were treated with different concentrations of SDS for one week. SDS affected duckweed enzymatic activities and phenol content. While in the fern phenolics amount, PAL, G-POD and PPO activities were not affected by SDS except for 100 ppm SDS, the only concentration that was taken up and not completely degraded. Stress ethylene production was induced only in the fern treated with 50 and 100 ppm SDS. Copyright © 2011 Elsevier B.V. All rights reserved.

  20. Superiority of SDS lysis over saponin lysis for direct bacterial identification from positive blood culture bottle by MALDI-TOF MS.

    PubMed

    Caspar, Yvan; Garnaud, Cécile; Raykova, Mariya; Bailly, Sébastien; Bidart, Marie; Maubon, Danièle

    2017-05-01

    Fast species diagnosis has an important health care impact, as rapid and specific antibacterial therapy is of clear benefit for patient's outcome. Here, a new protocol for species identification directly from positive blood cultures is proposed. Four in-house protocols for bacterial identification by MS directly from clinical positive blood cultures evaluating two lytic agents, SDS and saponin, and two protein extraction schemes, fast (FP) and long (LP) are compared. One hundred and sixty-eight identification tests are carried out on 42 strains. Overall, there are correct identifications to the species level in 90% samples for the SDS-LP, 60% for the SDS-FP, 48% for the saponin LP, and 43% for the saponin FP. Adapted scores allowed 92, 86, 72, and 53% identification for SDS-LP, SDS-FP, saponin LP, and saponin FP, respectively. Saponin lysis is associated with a significantly lower score compared to SDS (0.87 [0.83-0.92], p-value < 0.001). This study supports the use of SDS lysis instead of saponin lysis and the application of this rapid and cost-effective protocol in daily routine for microbiological agents implicated in septicemia. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Colorimetric protein determination in microalgae (Chlorophyta): association of milling and SDS treatment for total protein extraction.

    PubMed

    Mota, Maria Fernanda S; Souza, Marcella F; Bon, Elba P S; Rodrigues, Marcoaurelio A; Freitas, Suely Pereira

    2018-05-24

    The use of colorimetric methods for protein quantification in microalgae is hindered by their elevated amounts of membrane-embedded intracellular proteins. In this work, the protein content of three species of microalgae was determined by the Lowry method after the cells were dried, ball-milled, and treated with the detergent sodium dodecyl sulfate (SDS). Results demonstrated that the association of milling and SDS treatment resulted in a 3- to 7-fold increase in protein quantification. Milling promoted microalgal disaggregation and cell wall disruption enabling access of the SDS detergent to the microalgal intracellular membrane proteins and their efficient solubilization and quantification. © 2018 Phycological Society of America.

  2. Study of the interaction of proteins with curcumin and SDS and its analytical application

    NASA Astrophysics Data System (ADS)

    Wang, Feng; Yang, Jinghe; Wu, Xia; Liu, Shufang

    2005-09-01

    It is found that protein and sodium dodecyl sulphonate (SDS) can enhance resonance light scattering (RLS) of curcumin (CU). Based on this phenomenon, a new quantitative method for protein in aqueous solution has been developed. In the BR (pH 3.5) buffer, the RLS intensity of CU-SDS system is greatly enhanced by protein. The enhanced RLS is proportional to the concentration of protein in the range of 0.00020-20.0 μg ml -1 for bovine serum albumin (BSA) and 0.00040-1.0 μg ml -1 for human serum albumin (HSA) and their detection limits are 0.16 and 0.041 ng ml -1, respectively. An actual sample is satisfactorily determined. In addition, the interaction mechanism between protein and CU-SDS is also studied by using multi-techniques such as RLS, absorption spectroscopy and fluorescence, zeta potential assay measurement.

  3. Rapid recovery of dilute copper from a simulated Cu-SDS solution with low-cost steel wool cathode reactor.

    PubMed

    Chang, Shih-Hsien; Wang, Kai-Sung; Hu, Pei-I; Lui, I-Chun

    2009-04-30

    Copper-surfactant wastewaters are often encountered in electroplating, printed circuit boards manufacturing, and metal finishing industries, as well as in retentates from micellar-enhanced ultrafiltration process. A low-cost three-dimensional steel wool cathode reactor was evaluated for electrolytic recovery of Cu ion from dilute copper solution (0.2mM) in the presence of sodium dodecyl sulfate (SDS), octylphenol poly (ethyleneglycol) 9.5 ether (TX), nonylphenol poly (oxyethylene) 9 ether (NP9) and polyoxyethylene (20) sorbitan monooleate (TW) and also mixed surfactants (anionic/nonionic). The reactor showed excellent copper recovery ability in comparison to a parallel-plate reactor. The reactor rapidly recovered copper with a reasonable current efficiency. 93% of copper was recovered at current density of 1 A m(-2) and pH 4 in the presence of 8.5mM SDS. Initial solution pH, cathodic current density, solution mixing condition, SDS concentration, and initial copper concentrations significantly influenced copper recovery. The copper recovery rate increased with an increase in aqueous SDS concentrations between 5 and 8.5mM. The influences of nonionic surfactants on Cu recovery from SDS-Cu solution depended not only on the type of surfactants used, but also on applied concentrations. From the copper recovery perspective, TX at 0.1mM or NP should be selected rather than TW, because they did not inhibit copper recovery from SDS-Cu solution.

  4. Impact of the underlying etiology of growth hormone deficiency on serum IGF-I SDS levels during GH treatment in children.

    PubMed

    Léger, Juliane; Mohamed, Damir; Dos Santos, Sophie; Ben Azoun, Myriam; Zénaty, Delphine; Simon, Dominique; Paulsen, Anne; Martinerie, Laetitia; Chevenne, Didier; Alberti, Corinne; Carel, Jean-Claude; Guilmin-Crepon, Sophie

    2017-09-01

    Regular monitoring of serum IGF-I levels during growth hormone (GH) therapy has been recommended, for assessing treatment compliance and safety. To investigate serum IGF-I SDS levels during GH treatment in children with GH deficiency, and to identify potential determinants of these levels. This observational cohort study included all patients ( n  = 308) with childhood-onset non-acquired or acquired GH deficiency (GHD) included in the database of a single academic pediatric care center over a period of 10 years for whom at least one serum IGF-I SDS determination during GH treatment was available. These determinations had to have been carried out centrally, with the same immunoradiometric assay. Serum IGF-I SDS levels were determined as a function of sex, age and pubertal stage, according to our published normative data. Over a median of 4.0 (2-5.8) years of GH treatment per patient, 995 serum IGF-I SDS determinations were recorded. In addition to BMI SDS, height SDS and GH dose ( P  < 0.01), etiological group ( P  < 0.01) had a significant effect on serum IGF-I SDS levels, with patients suffering from acquired GHD having higher serum IGF-I SDS levels than those with non-acquired GHD, whereas sex, age, pubertal stage, treatment duration, hormonal status (isolated GHD (IGHD) vs multiple pituitary hormone deficiency (MPHD)) and initial severity of GHD, had no effect. These original findings have important clinical implications for long-term management and highlight the need for careful and appropriate monitoring of serum IGF-I SDS and GH dose, particularly in patients with acquired GHD, to prevent the unnecessary impact of potential comorbid conditions. © 2017 European Society of Endocrinology.

  5. Comparison of EDTA and SDS as potential surface impregnation agents for lead adsorption by activated carbon

    NASA Astrophysics Data System (ADS)

    Chen, Wei-fang; Pan, Ling; Chen, Li-fang; Yu, Zhe; Wang, Qiong; Yan, Chang-cheng

    2014-08-01

    Ethylene diamine tetraacetic acid (EDTA) and sodium dodecyl sulfate (SDS) were employed to impregnate activated carbons for the purpose of lead removal. The mechanisms of surface impregnation and lead adsorption method of chemical regeneration were investigated. Results showed that the highest impregnation of EDTA and SDS on activated carbon was 0.33 and 0.96 mmol/g, respectively. Adsorption capacities for lead of EDTA and SDS impregnated activated carbons reached 0.29 and 0.24 mmol/g. Rapid small scale column tests of adsorption and regeneration were conducted. Lead adsorption was greatly enhanced by EDTA impregnation. In addition, EDTA impregnated adsorbent was able to be successful regenerated by HNO3 and thus reused.

  6. Vertical two-phase flow regimes and pressure gradients under the influence of SDS surfactant

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Duangprasert, Tanabordee; Sirivat, Anuvat; Siemanond, Kitipat

    2008-01-15

    Two-phase gas/liquid flows in vertical pipes have been systematically investigated. Water and SDS surfactant solutions at various concentrations were used as the working fluids. In particular, we focus our work on the influence of surfactant addition on the flow regimes, the corresponding pressure gradients, and the bubble sizes and velocity. Adding the surfactant lowers the air critical Reynolds numbers for the bubble-slug flow and the slug flow transitions. The pressure gradients of SDS solutions are lower than those of pure water especially in the slug flow and the slug-churn flow regimes, implying turbulent drag reduction. At low Re{sub air}, themore » bubble sizes of the surfactant solution are lower than those of pure water due to the increase in viscosity. With increasing and at high Re{sub air}, the bubble sizes of the SDS solution become greater than those of pure water which is attributed to the effect of surface tension. (author)« less

  7. Temperature dependency of double material gate oxide (DMGO) symmetric dual-k spacer (SDS) wavy FinFET

    NASA Astrophysics Data System (ADS)

    Pradhan, K. P.; Priyanka; Sahu, P. K.

    2016-01-01

    Symmetric Dual-k Spacer (SDS) Trigate Wavy FinFET is a novel hybrid device that combines three significant and advanced technologies i.e., ultra-thin-body (UTB), FinFET, and symmetric spacer engineering on a single silicon on insulator (SOI) platform. This innovative architecture promises to enhance the device performance as compared to conventional FinFET without increasing the chip area. For the first time, we have incorporated two different dielectric materials (SiO2, and HfO2) as gate oxide to analyze the effect on various performance metrics of SDS wavy FinFET. This work evaluates the response of double material gate oxide (DMGO) on parameters like mobility, on current (Ion), transconductance (gm), transconductance generation factor (TGF), total gate capacitance (Cgg), and cutoff frequency (fT) in SDS wavy FinFET. This work also reveals the presence of biasing point i.e., zero temperature coefficient (ZTC) bias point. The ZTC bias point is that point where the device parameters become independent of temperature. The impact of operating temperature (T) on above said various performances are also subjected to extensive analysis. This further validates the reliability of DMGO-SDS FinFET and its application opportunities involved in modeling analog/RF circuits for a broad range of temperature applications. From extensive 3-D device simulation, we have determined that the inclusion of DMGO in SDS wavy FinFET is superior in performance.

  8. Removal of xylenol orange from its aqueous solution using SDS self-microemulsifying systems: optimization by Box-Behnken statistical design.

    PubMed

    Shakeel, Faiyaz; Haq, Nazrul; Alanazi, Fars K; Alsarra, Ibrahim A

    2014-04-01

    The aim of present study was to develop and evaluate sodium dodecyl sulfate (SDS) self-microemulsifying systems (SMES) for the removal of an anionic dye xylenol orange (XO) from its bulk aqueous media via liquid-liquid adsorption. The composition of SDS SMES was optimized by Box-Behnken statistical design for the maximum removal of XO from its aqueous solution. Various SDS formulations were prepared by spontaneous emulsification method and characterized for thermodynamic stability, self-microemulsification efficiency, droplet size, and viscosity. Adsorption studies were conducted at 8, 16, and 24 h by mixing small amounts of SDS formulations with relatively large amounts of bulk aqueous solution of XO. Droplet size and viscosity of SDS formulations were significantly influenced by oil phase concentration (triacetin), while surfactant concentration had little impact on droplet size and viscosity. However, the percentage of removal of XO was influenced by triacetin concentration, surfactant concentration, and adsorption time. Based on lowest droplet size (35.97 nm), lowest viscosity (29.62 cp), and highest percentage of removal efficiency (89.77 %), formulation F14, containing 2 % w/w of triacetin and 40 % w/w of surfactant mixture (20 % w/w of SDS and 20 % w/w of polyethylene glycol 400), was selected as an optimized formulation for the removal of XO from its bulk aqueous media after 16 h. These results indicated that SDS SMES could be suitable alternates of solid-liquid adsorption for the removal of toxic dyes such as XO from its aqueous solution through liquid-liquid adsorption.

  9. Evaluation of a combinatorial approach to prion inactivation using an oxidizing agent, SDS, and proteinase K.

    PubMed

    Smith, Jodi D; Nicholson, Eric M; Greenlee, Justin J

    2013-07-25

    Prions demonstrate an unusual resistance to methods effective at inactivating conventional microorganisms. This has resulted in a very tangible and difficult infection control challenge to the medical and veterinary communities, as well as animal agriculture and related industries. Currently accepted practices of harsh chemical treatments such as prolonged exposure to sodium hydroxide or sodium hypochlorite, or autoclaving are not suitable in many situations. Less caustic and more readily applicable treatments to contaminated environments are therefore desirable. We recently demonstrated that exposure of the RML scrapie agent to a commercial product containing sodium percarbonate (SPC-P) with or without sodium dodecyl sulfate (SDS) rendered PrP(Sc) sensitive to proteinase K (PK), but did not eliminate infectivity. The current study was designed to evaluate the efficacy of a combinatorial approach to inactivating prions by exposing RML-positive brain homogenate to SPC-P and SDS followed by PK. Treated samples were evaluated for PrP(Sc)-immunoreactivity by western blot, and residual infectivity by mouse bioassay. Treatment of infected brain homogenate with SPC-P and SDS followed by PK exposure resulted in a 4-5 log10 reduction in infectivity when bioassayed in tga20 mice. This study demonstrates that exposure of the RML scrapie agent to SPC-P and SDS followed by PK markedly reduces, but does not eliminate infectivity. The results of this study encourage further investigation into whether consecutive or concomitant exposure to sodium percarbonate, SDS, and a protease may serve as a viable and non-caustic option for prion inactivation.

  10. Gel electrophoretic isolation, in the hundred microgram range, of recombinant SDS-syntaxin from sea urchin egg cortical vesicles.

    PubMed

    Li, Y M; Chrambach, A

    2001-11-01

    %. S1, isolated from SDS-PAGE, exhibits an apparent Mr of 22.7 kDa, S2 one of 34.5 kDa, similar to the value of 32.6 kDa expected from the structure of syntaxin. The absence of S2 from the electroeluate re-electrophoresed at 0 degrees C and their molecular weight relationship suggest a proteolytic transformation of S2 to S1.

  11. Free radicals impair the anti-oxidative stress activity of DJ-1 through the formation of SDS-resistant dimer.

    PubMed

    Yasuda, Tatsuki; Niki, Takeshi; Ariga, Hiroyoshi; Iguchi-Ariga, Sanae M M

    2017-04-01

    DJ-1 is a causative gene for familial Parkinson's disease (PD). Loss-of-function of DJ-1 protein is suggested to contribute to the onset of PD, but the causes of DJ-1 dysfunction remain insufficiently elucidated. In this study, we found that the SDS-resistant irreversible dimer of DJ-1 protein was formed in human dopaminergic neuroblastoma SH-SY5Y cells when the cells were exposed to massive superoxide inducers such as paraquat and diquat. The dimer was also formed in vitro by superoxide in PQ redox cycling system and hydroxyl radical produced in Fenton reaction. We, thus, found a novel phenomenon that free radicals directly affect DJ-1 to form SDS-resistant dimers. Moreover, the formation of the SDS-resistant dimer impaired anti-oxidative stress activity of DJ-1 both in cell viability assay and H 2 O 2 -elimination assay in vitro. Similar SDS-resistant dimers were steadily formed with several mutants of DJ-1 found in familial PD patients. These findings suggest that DJ-1 is impaired due to the formation of SDS-resistant dimer when the protein is directly attacked by free radicals yielded by external and internal stresses and that the DJ-1 impairment is one of the causes of sporadic PD.

  12. Development of the Space Debris Sensor (SDS)

    NASA Technical Reports Server (NTRS)

    Hamilton, Joe; Liou, J. -C.; Anz-Meador, P.; Matney, M.; Christiansen, E.

    2017-01-01

    Debris Resistive/Acoustic Grid Orbital Navy-NASA Sensor (DRAGONS) is an impact sensor designed to detect and characterize collisions with small orbital debris: from 50 microns to greater than 1millimeter debris size detection; Characterizes debris size, speed, direction, and density. The Space Debris Sensor (SDS) is a flight demonstration of DRAGONS on the International Space Station: Approximately 1 square meter of detection area facing the ISS velocity vector; Minimum two year mission on Columbus External Payloads Facility (EPF); Minimal obstruction from ISS hardware; Development is nearing final checkout and integration with the ISS; Current launch schedule is SpaceX13, about September 2017, or SpaceX14, about Jan 2018.

  13. A Streamlined Western Blot Exercise: An Efficient and Greener Approach in the Laboratory Classroom

    ERIC Educational Resources Information Center

    Ness, Traci L.; Robinson, Rebekah L.; Mojadedi, Wais; Peavy, Lydia; Weiland, Mitch H.

    2015-01-01

    SDS-PAGE and western blotting are two commonly taught protein detection techniques in biochemistry and molecular biology laboratory classrooms. A pitfall associated with incorporating these techniques into the laboratory is the significant wait times that do not allow students to obtain timely results. The waiting associated with SDS-PAGE comes…

  14. Web Page Design.

    ERIC Educational Resources Information Center

    Lindsay, Lorin

    Designing a web home page involves many decisions that affect how the page will look, the kind of technology required to use the page, the links the page will provide, and kinds of patrons who can use the page. The theme of information literacy needs to be built into every web page; users need to be taught the skills of sorting and applying…

  15. Thermodynamics of Sodium Dodecyl Sulfate (SDS) Micellization: An Undergraduate Laboratory Experiment

    ERIC Educational Resources Information Center

    Marcolongo, Juan P.; Mirenda, Martin

    2011-01-01

    An undergraduate laboratory experiment is presented that allows a thermodynamic characterization of micelle formation of sodium dodecyl sulfate (SDS) in aqueous solutions. The critical micelle concentration (CMC) and the degree of micelle ionization (alpha) are obtained at different temperatures by conductimetry. The molar standard free energy…

  16. A Light-Responsive Self-Assembly Formed by a Cationic Azobenzene Derivative and SDS as a Drug Delivery System

    NASA Astrophysics Data System (ADS)

    Geng, Shengyong; Wang, Yuzhu; Wang, Liping; Kouyama, Tsutomu; Gotoh, Toshiaki; Wada, Satoshi; Wang, Jin-Ye

    2017-01-01

    The structure of a self-assembly formed from a cationic azobenzene derivative, 4-cholesterocarbonyl-4‧-(N,N,N-triethylamine butyloxyl bromide) azobenzene (CAB) and surfactant sodium dodecyl sulfate (SDS) in aqueous solution was studied by cryo-TEM and synchrotron radiation small-angle X-ray scattering (SAXS). Both unilamellar and multilamellar vesicles could be observed. CAB in vesicles were capable to undergo reversible trans-to-cis isomerization upon UV or visible light irradiation. The structural change upon UV light irradiation could be catched by SAXS, which demonstrated that the interlamellar spacing of the cis-multilamellar vesicles increased by 0.2-0.3 nm. Based on this microstructural change, the release of rhodamine B (RhB) and doxorubicin (DOX) could be triggered by UV irradiation. When incubated NIH 3T3 cells and Bel 7402 cells with DOX-loaded CAB/SDS vesicles, UV irradiation induced DOX release decreased the viability of both cell lines significantly compared with the non-irradiated cells. The in vitro experiment indicated that CAB/SDS vesicles had high efficiency to deliver loaded molecules into cells. The in vivo experiment showed that CAB/SDS vesicles not only have high drug delivery efficiency into rat retinas, but also could maintain high drug concentration for a longer time. CAB/SDS catanionic vesicles may find potential applications as a smart drug delivery system for controlled release by light.

  17. Foam capacity and stability of Sodium Dodecyl Sulfate (SDS) on the presence of contaminant coffee and Cd ions in solution

    NASA Astrophysics Data System (ADS)

    Haryanto, B.; Chang, C. H.; Kuo, A. T.; Siswarni, M. Z.; Sinaga, T. M. A.

    2018-02-01

    In this study, the effect of the coffee colloidal particle and Cd ion contaminant on the foam capacity and stability of sodium dodecyl sulfate (SDS) solution was investigated. The foam was generated by using a foam generator. The foam capacity of SDS was first evaluated at different concentrations. After the foam capacity reaching a constant value, the foam stability was then measured by flowing to a column. The results showed that the presence the coffee colloidal particles or Cd ions in the solution would decrease the foam capacity and stability of SDS. In addition, the decreased foam capacity and stability was more pronounced in the presence of coffee colloidal particles than Cd ions. The colloidal particles may have stronger interaction with SDS and thus reduce the formation of the foam.

  18. Quantitation of yeast total proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis sample buffer for uniform loading.

    PubMed

    Sheen, Hyukho

    2016-04-01

    Proteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) sample buffer are difficult to quantitate due to SDS and reducing agents being in the buffer. Although acetone precipitation has long been used to clean up proteins from detergents and salts, previous studies showed that protein recovery from acetone precipitation varies from 50 to 100% depending on the samples tested. Here, this article shows that acetone precipitates proteins highly efficiently from SDS-PAGE sample buffer and that quantitative recovery is achieved in 5 min at room temperature. Moreover, precipitated proteins are resolubilized with urea/guanidine, rather than with SDS. Thus, the resolubilized samples are readily quantifiable with Bradford reagent without using SDS-compatible assays. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. Interaction of SDS with β-galactosidase. A FT-IR study of the influence of detergent concentration and temperature.

    NASA Astrophysics Data System (ADS)

    Muga, A.; Castresana, J.; Arrondo, J. L. R.; López, S.; Bernabeu, C.

    1988-05-01

    The major structure of the enzyme β-galactosidase as studied by FT-IR is β-sheet with maxima in the amide I band at 1639 and 1655 cm -1 in H 2O and 1634 in D 2O. α-helix structure is also present with contribution from β-turns and less-ordered structure. Temperature induces a rearrangement of the structure producing a β-sheet-like conformation. In the presence of the surfactant SDS no big difference in structure is seen at 1% SDS (w:ww) concentration but there is a decrease of 5°C in the midpoint thermal denaturation. In the presence of 10% SDS a different picture is obtained with a higher random structure content.

  20. Polyurethane foam loaded with SDS for the adsorption of cationic dyes from aqueous medium: Multivariate optimization of the loading process.

    PubMed

    Robaina, Nicolle F; Soriano, Silvio; Cassella, Ricardo J

    2009-08-15

    This paper reports the development of a new procedure for the adsorption of four cationic dyes (Rhodamine B, Methylene Blue, Crystal Violet and Malachite Green) from aqueous medium employing polyurethane foam (PUF) loaded with sodium dodecylsulfate (SDS) as solid phase. PUF loading process was based on the stirring of 200mg PUF cylinders with acidic solutions containing SDS. The conditions for loading were optimized by response surface methodology (RSM) using a Doehlert design with three variables that were SDS and HCl concentrations and stirring time. Results obtained in the optimization process showed that the stirring time is not a relevant parameter in the PUF loading, evidencing that the transport of SDS from solution to PUF surface is fast. On the other hand, both SDS and HCl concentrations were important parameters causing significant variation in the efficiency of the resulting solid phase for the removal of dyes from solution. At optimized conditions, SDS and HCl concentrations were 4.0 x 10(-4) and 0.90 mol L(-1), respectively. The influence of stirring time was evaluated by univariate methodology. A 20 min stirring time was established in order to make the PUF loading process fast and robust without losing efficiency. The procedure was tested for the removal of the four cationic dyes from aqueous solutions and removal efficiencies always better than 90% were achieved for the two concentrations tested (2.0 x 10(-5) and 1.0 x 10(-4)mol L(-1)).

  1. Removal of fluoride, SDS, ammonia and turbidity from semiconductor wastewater by combined electrocoagulation-electroflotation.

    PubMed

    Aoudj, S; Khelifa, A; Drouiche, N

    2017-08-01

    Semiconductor industry effluents contain organic and inorganic pollutants, such as sodium dodecyl sulfate (SDS), fluoride and ammonia, at high levels which consists a major environmental issue. A combined EC-EF process is proposed as a post-treatment after precipitation for simultaneous clarification and removal of pollutants. In EC step, a hybrid Fe-Al was used as the soluble anode in order to avoid supplementary EC step. EC-Fe is more suitable for SDS removal; EC-Al is more suitable for fluoride removal, while EC with hybrid Al-Fe makes a good compromise. Clarification and ammonia oxidation were achieved in the EF step. Effects of anodic material, initial pH, current, anion nature, chloride concentration and initial pollutant concentration were studied. The final concentrations may reach 0.27, 6.23 and 0.22 mg L -1 for SDS, fluoride and ammonia respectively. These concentrations are far lower than the correspondent discharge limits. Similarly, the final turbidity was found 4.35 NTU which is lower than 5NTU and the treated water does not need further filtration before discharge. Furthermore, the EC-EF process proves to be sufficiently energy-efficient with less soluble electrode consumption. Copyright © 2017 Elsevier Ltd. All rights reserved.

  2. SDS-binding assay based on tyrosine fluorescence as a tool to determine binding properties of human serum albumin in blood plasma

    NASA Astrophysics Data System (ADS)

    Zhdanova, Nadezda; Shirshin, Evgeny; Fadeev, Victor; Priezzhev, Alexander

    2016-04-01

    Among all plasma proteins human serum albumin (HSA) is the most studied one as it is the main transport protein and can bind a wide variety of ligands especially fatty acids (FAs). The concentration of FAs bound to HSA in human blood plasma differs by three times under abnormal conditions (fasting, physical exercises or in case of social important diseases). In the present study a surfactant sodium dodecyl sulfate (SDS) was used to simulate FAs binding to HSA. It was shown that the increase of Tyr fluorescence of human blood plasma due to SDS addition can be completely explained by HSA-SDS complex formation. Binding parameters of SDS-HSA complex (average number of sites and apparent constant of complex formation) were determined from titration curves based on tyrosine (Tyr) fluorescence.

  3. Quantitative trait loci underlying resistance to sudden death syndrome (SDS) in MD96-5722 by 'Spencer' recombinant inbred line population of soybean.

    PubMed

    Anderson, J; Akond, M; Kassem, M A; Meksem, K; Kantartzi, S K

    2015-04-01

    The best way to protect yield loss of soybean [Glycine max (L.) Merr.] due to sudden death syndrome (SDS), caused by Fusarium virguliforme (Aoki, O'Donnel, Homma & Lattanzi), is the development and use of resistant lines. Mapping quantitative trait loci (QTL) linked to SDS help developing resistant soybean germplasm through molecular marker-assisted selection strategy. QTL for SDS presented herein are from a high-density SNP-based genetic linkage map of MD 96-5722 (a.k.a 'Monocacy') by 'Spencer' recombinant inbred line using SoySNP6K Illumina Infinium BeadChip genotyping array. Ninety-four F 5:7 lines were evaluated for 2 years (2010 and 2011) at two locations (Carbondale and Valmeyer) in southern Illinois, USA to identify QTL controlling SDS resistance using disease index (DX). Composite interval mapping identified 19 SDS controlling QTL which were mapped on 11 separate linkage group (LG) or chromosomes (Chr) out of 20 LG or Chr of soybean genome. Many of these significant QTL identified in one environment/year were confirmed in another year or environment, which suggests a common genetic effects and modes of the pathogen. These new QTL are useful sources for SDS resistance studies in soybean breeding, complementing previously reported loci.

  4. Influence of water-insoluble nonionic copolymer E(6)P(39)E(6) on the microstructure and self-aggregation dynamics of aqueous SDS solution-NMR and SANS investigations.

    PubMed

    Prameela, G K S; Phani Kumar, B V N; Aswal, V K; Mandal, Asit Baran

    2013-10-28

    The influence of water-insoluble nonionic triblock copolymer PEO-PPO-PEO [poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide)] i.e., E6P39E6 with molecular weight 2800, on the microstructure and self-aggregation dynamics of anionic surfactant sodium dodecylsulfate (SDS) in aqueous solution (D2O) were investigated using high resolution nuclear magnetic resonance (NMR) and small-angle neutron scattering (SANS) measurements. Variable concentration and temperature proton ((1)H), carbon ((13)C) NMR chemical shifts, (1)H self-diffusion coefficients, (1)H spin-lattice and spin-spin relaxation rates data indicate that the higher hydrophobic nature of copolymer significantly influenced aggregation characteristics of SDS. The salient features of the NMR investigations include (i) the onset of mixed micelles at lower SDS concentrations (<3 mM) relative to the copolymer-free case and their evolution into SDS free micelles at higher SDS concentrations (~30 mM), (ii) disintegration of copolymer-SDS mixed aggregate at moderate SDS concentrations (~10 mM) and still binding of a copolymer with SDS and (iii) preferential localization of the copolymer occurred at the SDS micelle surface. SANS investigations indicate prolate ellipsoidal shaped mixed aggregates with an increase in SDS aggregation number, while a contrasting behavior in the copolymer aggregation is observed. The aggregation features of SDS and the copolymer, the sizes of mixed aggregates and the degree of counterion dissociation (α) extracted from SANS data analysis corroborate reasonably well with those of (1)H NMR self-diffusion and sodium ((23)Na) spin-lattice relaxation data.

  5. iSDS: a self-configurable software-defined storage system for enterprise

    NASA Astrophysics Data System (ADS)

    Chen, Wen-Shyen Eric; Huang, Chun-Fang; Huang, Ming-Jen

    2018-01-01

    Storage is one of the most important aspects of IT infrastructure for various enterprises. But, enterprises are interested in more than just data storage; they are interested in such things as more reliable data protection, higher performance and reduced resource consumption. Traditional enterprise-grade storage satisfies these requirements at high cost. It is because traditional enterprise-grade storage is usually designed and constructed by customised field-programmable gate array to achieve high-end functionality. However, in this ever-changing environment, enterprises request storage with more flexible deployment and at lower cost. Moreover, the rise of new application fields, such as social media, big data, video streaming service etc., makes operational tasks for administrators more complex. In this article, a new storage system called intelligent software-defined storage (iSDS), based on software-defined storage, is described. More specifically, this approach advocates using software to replace features provided by traditional customised chips. To alleviate the management burden, it also advocates applying machine learning to automatically configure storage to meet dynamic requirements of workloads running on storage. This article focuses on the analysis feature of iSDS cluster by detailing its architecture and design.

  6. [Construction of recombinant adenovirus and mediated reported gene expression in the guinea pig cochlea].

    PubMed

    Liu, Yingpeng; Wang, Guopeng; Shen, Anmin; Wang, Jianting; Chen, Pei; Li, Zeweng; Gong, Shusheng

    2007-08-01

    To purify P0 protein from guinea pig's inner ear by preparative SDS-PAGE and study the possible role it may play in the etiology of autoimmune inner ear disease. A mixture of membraneous proteins of inner ear was separated by preparative SDS-PAGE. The corresponding band at 30kd was cut and electrically eluted. The protein collected was identified by analytical SDS-PAGE and Western blot assay. A group of 20 guinea pigs were immunized with P0 protein emulsified in complete Freund's adjuvant, another 10 guinea pigs were immunized with complete Freund 's adjuvant only as control. The guinea pigs' hearing thresholds, serum IgG level and morphological changes in the inner ear were investigated. The distribution of P0 protein in the cochlear was detected by immunohistochemical technique. The purity of the protein was demonstrated by a single band at the 30 kD site in SDS-PAGE, which was identified as P0 protein by western blot analysis assay. About 17.5% P0-immunized guinea pigs showed increased hearing thresholds, elevated IgG level (F =6.48, P <0. 01), as well as a decreased number of spiral ganglion cells and inflammatory cell infiltration in the cochlear nerve region. The P0 protein is distributed in the cochlear nerve and spiral ganglion only. P0 protein from guinea pig's inner ear can be successfully purified by preparative SDS-PAGE and an animal model of experimental autoimmune inner ear disease induced by P0 protein is successfully established.

  7. Comparison and validation of methods to quantify Cry1Ab toxin from Bacillus thuringiensis for standardization of insect bioassays.

    PubMed

    Crespo, André L B; Spencer, Terence A; Nekl, Emily; Pusztai-Carey, Marianne; Moar, William J; Siegfried, Blair D

    2008-01-01

    Standardization of toxin preparations derived from Bacillus thuringiensis (Berliner) used in laboratory bioassays is critical for accurately assessing possible changes in the susceptibility of field populations of target pests. Different methods were evaluated to quantify Cry1Ab, the toxin expressed by 80% of the commercially available transgenic maize that targets the European corn borer, Ostrinia nubilalis (Hübner). We compared three methods of quantification on three different toxin preparations from independent sources: enzyme-linked immunosorbent assay (ELISA), sodium dodecyl sulfate-polyacrylamide gel electrophoresis and densitometry (SDS-PAGE/densitometry), and the Bradford assay for total protein. The results were compared to those obtained by immunoblot analysis and with the results of toxin bioassays against susceptible laboratory colonies of O. nubilalis. The Bradford method resulted in statistically higher estimates than either ELISA or SDS-PAGE/densitometry but also provided the lowest coefficients of variation (CVs) for estimates of the Cry1Ab concentration (from 2.4 to 5.4%). The CV of estimates obtained by ELISA ranged from 12.8 to 26.5%, whereas the CV of estimates obtained by SDS-PAGE/densitometry ranged from 0.2 to 15.4%. We standardized toxin concentration by using SDS-PAGE/densitometry, which is the only method specific for the 65-kDa Cry1Ab protein and is not confounded by impurities detected by ELISA and Bradford assay for total protein. Bioassays with standardized Cry1Ab preparations based on SDS-PAGE/densitometry showed no significant differences in LC(50) values, although there were significant differences in growth inhibition for two of the three Cry1Ab preparations. However, the variation in larval weight caused by toxin source was only 4% of the total variation, and we conclude that standardization of Cry1Ab production and quantification by SDS-PAGE/densitometry may improve data consistency in monitoring efforts to identify changes in

  8. Transfer in SDS of biotinylated proteins from acrylamide gels to an avidin-coated membrane filter.

    PubMed

    Karlin, Arthur; Wang, Chaojian; Li, Jing; Xu, Qiang

    2004-06-01

    Avidin was covalently linked to aldehyde-derivatized polyethersulfone membrane filters. These filters were used in Western blot analysis of proteins reacted with biotinylation reagents and electrophoresed in sodium dodecyl sulfate (SDS) on polyacrylamide gels. Electrophoretic transfer from the gels to these filters was in 0.1% SDS, in which the covalently bound avidin retained its biotin-binding capacity. We compared Western blots on avidin-coated membrane filters of biotinylated and nonbiotinylated forms of mouse immunoglobulin G (IgG), mouse IgG heavy chain, muscle-type acetylcholine receptor alpha subunit, and fused alpha and beta subunits of receptor. Biotinylated proteins were captured with high specificity compared to their nonbiotinylated counterparts and sensitively detected on the avidin-coated membranes.

  9. Molecular Level Understanding of Sodium Dodecyl Sulfate (SDS) Induced Sol-Gel Transition of Pluronic F127 Using Fisetin as a Fluorescent Molecular Probe.

    PubMed

    Mishra, Jhili; Swain, Jitendriya; Mishra, Ashok Kumar

    2018-01-11

    The thermoreversible sol-gel transition of pluronic F127 is markedly altered even with addition of submicellar concentration of sodium dodecyl sulfate (SDS) surfactant. Multiple fluorescence parameters like fluorescence intensity, fluorescence anisotropy and fluorescence lifetime of both the prototropic forms (anion (A - *) and phototautomer FT*) of the photoprototropic fluorescent probe fisetin has been efficiently used to understand the molecular level properties like polarity and microviscosity of the PF127-SDS system as a function of temperature. The SDS-induced increase in the interfacial hydrophobicity level is seen to affect the sol-gel phase transition of PF127 (21-18 °C). The E T (30) polarity parameter value of anionic emission of fisetin suggests that there is a considerable decrease in the polarity of the PF127 medium with increase in temperature and with the addition of SDS. The microviscosity progressively increases from ∼5 mPa s (sol state, 10 °C) to ∼22.01 mPa s (gel state 35 °C) in aqueous solution of PF127. The variation in microviscosity with addition of SDS in PF127-SDS mixed system is significant in sol phase whereas in gel phase this variation is significantly less. Temperature dependent fluorescence lifetime of FT* indicates that there is heterogeneity in distribution of fisetin molecules at different domains of PF127. This work also show-cases the sensitivity of fisetin toward change in polarity and change in sol-gel transition temperature of copolymer PF127 with variation in temperature (both forward and reverse directions) and SDS.

  10. Thermodynamics of Interaction between Some Cellulose Ethers and SDS by Titration Microcalorimetry.

    PubMed

    Singh; Nilsson

    1999-05-01

    The interaction between certain nonionic cellulose ethers (ethyl hydroxyethyl cellulose and hydroxypropyl methyl cellulose) and sodium dodecyl sulphate (SDS) has been investigated using isothermal titration microcalorimetry at temperatures between 25-50 degrees C. The observed heat flow curves have been interpreted in terms of a plausible mechanism of the interaction of the substituent groups with SDS monomers and clusters. The data have been related to changes occuring in the system at the macro- and microscopic levels with the addition of surfactants and with temperature. The process consists predominantly of polymer-surfactant interactions initially and surfactant-surfactant interactions at the later stages. A phenomenological model of the cooperative interaction (adsorption) process has been derived, and earlier published equilibrium binding data have been used to recover binding constants and Gibbs energy changes for this process. The adsorption enthalpies and entropies have been recovered along with the heat capacity change. The enthalpic cost of confining the nonpolar regions of the polymers in surfactant clusters is high, but the entropy gain from release of hydration shell water molecules as well as increased freedom of movement of these nonpolar regions in the clusters gives the process a strong entropic driving force. The process is entropy-driven initially and converts to being both enthalpy and entropy-driven at high SDS concentrations. An enthalpy-entropy compensation behavior is seen. Strongly negative heat capacity changes have been obtained resulting from the transfer of nonpolar groups from aqueous into nonpolar environments, as well as a reduction of conformational domains that the chains can populate. Changes in these two components cause the heat capacity change to become less negative at the higher binding levels. The system can be classified as exhibiting nonclassical hydrophobic binding at the later stages of binding. Copyright 1999 Academic

  11. Product yield-detected ESR on magnetic field-dependent photoreduction of quinones in SDS micellar solution

    NASA Astrophysics Data System (ADS)

    Okazaki, M.; Sakata, S.; Konaka, R.; Shiga, T.

    1987-06-01

    Transient free radicals in the magnetic field-dependent photoreduction of quinones (menadione or anthraquinone) in a sodium dodecyl sulfate (SDS) micellar solution, were converted to stable nitroxide radicals by the ``spin trapping'' technique with or without the microwave irradiation. Upon irradiating the microwave at 160 mW, the product yield (``spin adduct'' of the alkyl radical generated from SDS molecule) decreased by up to 14% at certain magnetic fields in a resonant manner. Although only one component of the postulated radical pair was converted to the spin adduct, the decrease in the yield as a function of external magnetic field revealed the ESR spectra of both component radicals of the radical pair, i.e., the semiquinone radical and the alkyl radical from SDS. This experiment not only gives the direct evidence for the radical pair model, but also suggests the possibility for this method to be applied in controlling the chemical reactions by the microwave. A simple calculation was made to simulate the observed ``product yield-detected ESR.'' Agreements were achieved semiquantitatively between the observed reductions in the spin adduct yields and those calculated. The estimated exchange interaction between the component radicals in the radical pair of the present systems was lower than 0.3 mT.

  12. SLAC Detailed Page: For staff, users, and collaborators - Page no longer

    Science.gov Websites

    information about this change.) This page will automatically redirect to the For Staff page. You may also want to visit the new Detailed Index web page. Please change your bookmarks accordingly. SLAC Stanford

  13. Pterostilbene, a Potent Analog of Resveratrol, a Therapeutic Agent in Prostate Cancer: Epigenetic Mechanisms of Action

    DTIC Science & Technology

    2015-10-01

    protein was loaded in 10-15% SDS-PAGE and transferred onto polyvinylidene difluoride (PVDF) membranes. Membranes were incubated in 5% nonfat dry milk ...epigenetic reader, metastasis-associated protein 1 (MTA1) which is a part of nucleosome remodeling and deacetylation (NuRD) co-repressor complex. MTA1...inhibitor cocktail (ThermoFisher Scientific). Samples containing 70 μg of protein were loaded in 10-12% SDS-PAGE and transferred onto polyvinylidene

  14. Using Two Different Self-Directed Search (SDS) Interpretive Materials: Implications for Career Assessment

    ERIC Educational Resources Information Center

    Dozier, V. Casey; Sampson, James P.; Reardon, Robert C.

    2013-01-01

    John Holland's Self-Directed Search (SDS) is a career assessment that consists of several booklets designed to be self-scored and self-administered. It simulates what a practitioner and an individual might do together in a career counseling session (e.g., review preferred activities and occupations; review competencies, abilities and possible…

  15. Congruency between Occupational Daydreams and Self Directed Search (SDS) Scores among College Students

    ERIC Educational Resources Information Center

    Miller, Mark J.; Springer, Thomas P.; Tobacyk, Jerome; Wells, Don

    2004-01-01

    In this study, the relationship of expressed occupational daydreams and scores on the Self-Directed Search (SDS) were examined. Results were consistent with Holland's theory of careers. Implications for career counselors are discussed. Students were asked to provide specific biographical data (i. e., age, gender, race) and to write down their…

  16. Page turning system

    NASA Technical Reports Server (NTRS)

    Kerley, James J. (Inventor); Eklund, Wayne D. (Inventor)

    1992-01-01

    A device for holding reading materials for use by readers without arm mobility is presented. The device is adapted to hold the reading materials in position for reading with the pages displayed to enable turning by use of a rubber tipped stick that is held in the mouth and has a pair of rectangular frames. The frames are for holding and positioning the reading materials opened in reading posture with the pages displayed at a substantially unobstructed sighting position for reading. The pair of rectangular frames are connected to one another by a hinge so the angle between the frames may be varied thereby varying the inclination of the reading material. A pair of bent spring mounted wires for holding opposing pages of the reading material open for reading without substantial visual interference of the pages is mounted to the base. The wires are also adjustable to the thickness of the reading material and have a variable friction adjustment. This enables the force of the wires against the pages to be varied and permits the reader to manipulate the pages with the stick.

  17. 8. Photocopy of printed page (original Page 30 of the ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    8. Photocopy of printed page (original Page 30 of the Souvenir Program 1867-1967 Ridgely Centennial) Photographer unknown. Circa 1967. VIEW NORTHEAST, SOUTHWEST FRONT Ridgely's centennial was celebrated in 1967 and included in the souvenir brochure was page 30. This view shows the subject building with the 1950 modifications to provide for automotive traffic. It was a print of a current photograph. - 510 Central Avenue (Commercial Building), Ridgely, Caroline County, MD

  18. Primordial SdS universe from a 5D vacuum: scalar field fluctuations on Schwarzschild and Hubble horizons

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Aguilar, José Edgar Madriz; Bellini, Mauricio, E-mail: jemadriz@fisica.ugto.mx, E-mail: mbellini@mdp.edu.ar

    2010-11-01

    We study scalar field fluctuations of the inflaton field in an early inflationary universe on an effective 4D Schwarzschild-de Sitter (SdS) metric, which is obtained after make a planar coordinate transformation on a 5D Ricci-flat Schwarzschild-de Sitter (SdS) static metric. We obtain the important result that the spectrum of fluctuations at zeroth order is independent of the scalar field mass M on Schwarzschild scales, while on cosmological scales it exhibits a mass dependence. However, in the first-order expansion, the spectrum depends of the inflaton mass and the amplitude is linear with the Black-Hole (BH) mass m.

  19. Purification and protein composition of endogenous rat viruses.

    PubMed

    Hlubinová, K; Prachar, J; Vrbenská, A; Matoska, J; Simkovic, D

    1984-01-01

    Endogenous retroviruses are not in the majority of cases the cause of any neoplasia, except for the laboratory conditions. As far as they might serve for the evolution of pathogenic retroviruses more attention should have been paid to them. In this paper we introduce some approaches to the purification of rat endogenous retroviruses to such a degree of purity that enabled satisfactory SDS-PAGE analysis of its structural proteins. Purities of samples obtained by usual purification methods, long-term isopycnic centrifugation at a high gravity force and velocity centrifugation are compared. Protein profile of rat endogenous virus in SDS-PAGE is compared with the ones of other retroviruses. For the first time the evidence was obtained for the striking similarity between electrophoretic protein profile of rat endogenous virus WERC and feline leukemia virus. The major structural proteins of rat endogenous retrovirus and feline leukemia virus cannot be distinguished even when resolution long gradient PAGE had been employed. The accordance of electrophoretic mobilities of major structural proteins in SDS-PAGE can indicate the relatedness of retroviruses.

  20. Holland's SDS Applied to Chinese College Students: A Revisit to Cross-Culture Adaptation

    ERIC Educational Resources Information Center

    Kong, Jin; Xu, Yonghong Jade; Zhang, Hao

    2016-01-01

    In this study, data collected from 875 college freshman and sophomore students enrolled in a 4-year university in central China are used to examine the applicability and validity of a Chinese version of Holland's Self-Directed Search (SDS) that was adapted in the 1990s. The total sample was randomly divided into two groups. Data from the first…

  1. Targeted Elimination of PCDH-PC Expressing Prostate Cancer Cells for Control of Hormone-Resistant Prostate Cancer

    DTIC Science & Technology

    2007-11-01

    SDS-PAGE gel . The Western blot made from this gel was probed with antibody that recognizes the myc-tag. When compared to the extracts from the...SDS-PAGE gel and blotted onto a filter. The filter was probed with an anti-myc antibody. The levels of myc-tagged PCDH-PC protein in cells co...Specific Aim 2. Design and test antisense oligonucleotides ( ASOs ) that suppress PCDH-PC expression in prostate cancer cells. Work Done: We used

  2. Problems with multiple use of transfer buffer in protein electrophoretic transfer.

    PubMed

    Dorri, Yaser; Kurien, Biji T; Scofield, R Hal

    2010-04-01

    Two-dimensional gel electrophoresis (2DE) and SDS-PAGE are the two most useful methods in protein separation. Proteins separated by 2DE or SDS-PAGE are usually transferred to membranes using a variety of methods, such as electrophoretic transfer, heat-mediated transfer, or nonelectrophoretic transfer, for specific protein detection and/or analysis. In a recent study, Pettegrew et al. claim to reuse transfer buffer containing methanol for at least five times for transferring proteins from SDS-PAGE to polyvinylidene difluoride. They add 150-200 ml fresh transfer solution each time for extended use as a result of loss of transfer buffer. Finally, they test efficiency of each protein transfer by chemiluminescence detection. Here, we comment on this report, as we believe this method is not accurate and useful for protein analysis, and it can cause background binding as well as inaccurate protein analysis.

  3. The NPOESS Preparatory Project Science Data Segment (SDS) Data Depository and Distribution Element (SD3E) System Architecture

    NASA Technical Reports Server (NTRS)

    Ho, Evelyn L.; Schweiss, Robert J.

    2008-01-01

    The National Polar-orbiting Operational Environmental Satellite System (NPOESS) Preparatory Project (NPP) Science Data Segment (SDS) will make daily data requests for approximately six terabytes of NPP science products for each of its six environmental assessment elements from the operational data providers. As a result, issues associated with duplicate data requests, data transfers of large volumes of diverse products, and data transfer failures raised concerns with respect to the network traffic and bandwidth consumption. The NPP SDS Data Depository and Distribution Element (SD3E) was developed to provide a mechanism for efficient data exchange, alleviate duplicate network traffic, and reduce operational costs.

  4. [Characterization of haemolysis of the Vibrio parahaemolyticus no.93].

    PubMed

    Su, S C; Lee, C Y

    1997-02-01

    Vibrio parahaemolyticus is a causative bacterium of food poisoning, and the haemolysin produced by this organism has been considered as one of the important virulence factors. In order to understand the pathogenic mechanism of this bacterium, the characteristics of haemolysin from Vibrio parahaemolyticus isolated from Taiwan were studied. One of the clinical strains, V. parahaemolyticus No.93, presents a weak hemolytic zone on 7% NaCl-Wagatsuma medium. The DNA hybridization results show that V. parahemolyticus has neither tdh nor trh gene. V. parahaemolyticus No.93 shows obviously hemolytic zone on 3%-NaCl Wagatsuma medium (human blood). The crude extracellular protein of V. parahaemolyticus No. 93 was evaluated for its heat tolerance and enzyme activities by media assay. The results show that this crude extracellular protein is thermolabile. The crude extracellular protein of V. parahaemolyticus No.93 was analyzed on 10% SDS-PAGE and an apparent band of 64 kDa protein was observed. Furthermore, the crude extracellular protein was analyzed by running gelatin-SDS-PAGE and hemoglobin-SDS-PAGE, and three clear zones on 62 kDa, 52 kDa and 41 kDa were observed on both SDS-PAGEs. Thus we propose that the crude extracellular protein of the V. parahaemolyticus No.93 can degrade gelatin as well as hemoglobin. Whether these protease being the virulence factors of Vibrio parahaemolyticus No.93 needs to be further studied.

  5. [Optimized isolation and purification of non-typeable Haemophilus influenzae Haps protein].

    PubMed

    Li, Wan-yi; Kuang, Yu; Li, Ming-yuan; Yang, Yuan; Jiang, Zhong-hua; Yao, Feng; Chen, Chang-chun

    2007-12-01

    To optimize the isolation and purification conditions for Hap(s) protein of non-typeable Haemophilus influenzae. Hap(s) protein was purified by ammonium sulfate precipitation, dialysis desalting and Hitrap weak cation exchange columns of CM Sepharose Fast Flow. The condition of the elution was optimized for pH and ionic strength, the absorbance at 280 nm of the elution samples were detected, and the targeted protein band in the collected samples was observed by SDS-PAGE electrophoresis. The Hitrap ion exchange column was eluted with buffer 1, which resulted in a baseline distribution of absorbance at 280 nm. Buffer 2 elution of the column resulted in the presence of peak absorbance with trails, which was identified to be constituted by some low molecular weight bands by subsequent SDS-PAGE. In serial column elution with buffer 3 with different ionic strength, a peak absorbance was observed with the ionic strength of 100 mmol/L NaCl, and SDS-PAGE confirmed that the peak was generated by the target protein. No obvious peaks or bands in SDS-PAGE occurred with the other ionic strengths. The pH of the buffer only affect the elution of the irrelevant proteins rather than the Hap(s) protein, and elution with the buffer containing 100 mmol/L NaCl can be optimal for eluting the Hap(s) protein.

  6. Lipids and proteins in the Rathke's gland secretions of the North American mud turtle (Kinosternon subrubrum)

    USGS Publications Warehouse

    Seifert, W.E.; Gotte, S.W.; Leto, T.L.; Weldon, P.J.

    1994-01-01

    Lipids and proteins in the Rathke's gland secretions of the North American mud turtle (Kinosternon subrubrum, Kinosternidae) were analyzed by gas chromatography-mass spectrometry (GC-MS) and SDS-polyacrylamide gel electrophoresis (SDS-PAGE), respectively. Analysis by GC-MS indicates 2,3-dihydroxypropanal and C3–C24 free or esterified fatty acids. Analysis by SDS-PAGE indicates a major protein component with an approximate molecular mass of 60 kDa and minor components ranging from ca. 23 to 34 kDa. The major component of K. subrubrum glandular secretions exhibits a mobility that matches that of the Kemp's ridley sea turtle (Lepidochelys kempi, Cheloniidae), suggesting that these proteins are evolutionarily conserved.

  7. SDS-MoS2 nanoparticles as highly-efficient peroxidase mimetics for colorimetric detection of H2O2 and glucose.

    PubMed

    Zhao, Kai; Gu, Wei; Zheng, Sisi; Zhang, Cuiling; Xian, Yuezhong

    2015-08-15

    In this work, we find that the peroxidase-like activity of MoS2 nanoparticles (NPs) is dependent on the surface charge. Negatively charged sodium dodecyl sulfate modified MoS2 nanoparticles (SDS-MoS2 NPs) possess highly-efficient peroxidase-like activity. MoS2 NPs with intrinsic peroxidase-like activity were synthesized through a simple one-pot hydrothermal route. The peroxidase-like activities of different surfactants modified MoS2 NPs were discussed. Compared with bare MoS2 NPs and positively charged cetyltrimethyl ammonium bromide modified MoS2 NPs, SDS-MoS2 NPs have the best peroxidase-like activity. SDS-MoS2 NPs can efficiently catalyze the oxidation of 3,3,5,5-tetramethylbenzidine (TMB) in the presence of H2O2 to generate a blue product. On basis of this, we have successfully established a novel platform for colorimetric detection of H2O2, and the detection limit is 0.32μM. Furthermore, the SDS-MoS2 NPs based platform can also be used for high sensitivity and selectivity detection of glucose with a wide linear range of 5.0-500μM by controlling the generation of H2O2 in the presence of glucose oxidase. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. The impact of N,N-dimethyldodecylamine N-oxide (DDAO) concentration on the crystallisation of sodium dodecyl sulfate (SDS) systems and the resulting changes to crystal structure, shape and the kinetics of crystal growth.

    PubMed

    Summerton, Emily; Hollamby, Martin J; Zimbitas, Georgina; Snow, Tim; Smith, Andrew J; Sommertune, Jens; Bettiol, Jeanluc; Jones, Christopher; Britton, Melanie M; Bakalis, Serafim

    2018-05-19

    At low temperatures stability issues arise in commercial detergent products when surfactant crystallisation occurs, a process which is not currently well-understood. An understanding of the phase transition can be obtained using a simple binary SDS (sodium dodecyl sulfate) + DDAO (N,N-dimethyldodecylamine N-oxide) aqueous system. It expected that the crystallisation temperature of an SDS system can be lowered with addition of DDAO, thus providing a route to improve detergent stability. Detergent systems are typically comprised of anionic surfactants, non-ionic surfactants and water. This study explores the crystallisation of a three component system consisting of sodium dodecyl sulfate (SDS), N,N-dimethyldodecylamine N-oxide (DDAO), and water using wide-angle X-ray scattering (WAXS), differential scanning calorimetry (DSC) and confocal Raman microscopy. The presence of DDAO lowered the crystallisation temperature of a 20 wt% SDS system. For all aqueous mixtures of SDS + DDAO at low temperatures, SDS hydrated crystals, SDS.1/2H 2 O or SDS·H 2 O, formed. SDS hydrates comprising of layers of SDS separated by water layers. DDAO tended to reside in the vicinity of these SDS crystals. In the absence of DDAO an additional intermediary hydrate structure, SDS.1/8H 2 O, formed whereas for mixed SDS + DDAO systems no such structure was detected during crystallisation. Copyright © 2018. Published by Elsevier Inc.

  9. Enhanced protein electrophoresis technique for separating human skeletal muscle myosin heavy chain isoforms

    NASA Technical Reports Server (NTRS)

    Bamman, M. M.; Clarke, M. S.; Talmadge, R. J.; Feeback, D. L.

    1999-01-01

    Talmadge and Roy (J. Appl. Physiol. 1993, 75, 2337-2340) previously established a sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SDS-PAGE) protocol for separating all four rat skeletal muscle myosin heavy chain (MHC) isoforms (MHC I, IIa, IIx, IIb); however, when applied to human muscle, the type II MHC isoforms (Ila, IIx) are not clearly distinguished. In this brief paper we describe a modification of the SDS-PAGE protocol which yields distinct and consistent separation of all three adult human MHC isoforms (MHC I, IIa, IIx) in a minigel system. MHC specificity of each band was confirmed by Western blot using three monoclonal IgG antibodies (mAbs) immunoreactive against MHCI (mAb MHCs, Novacastra Laboratories), MHCI+IIa (mAb BF-35), and MHCIIa+IIx (mAb SC-71). Results provide a valuable SDS-PAGE minigel technique for separating MHC isoforms in human muscle without the difficult task of casting gradient gels.

  10. The Molecule Pages database

    PubMed Central

    Saunders, Brian; Lyon, Stephen; Day, Matthew; Riley, Brenda; Chenette, Emily; Subramaniam, Shankar

    2008-01-01

    The UCSD-Nature Signaling Gateway Molecule Pages (http://www.signaling-gateway.org/molecule) provides essential information on more than 3800 mammalian proteins involved in cellular signaling. The Molecule Pages contain expert-authored and peer-reviewed information based on the published literature, complemented by regularly updated information derived from public data source references and sequence analysis. The expert-authored data includes both a full-text review about the molecule, with citations, and highly structured data for bioinformatics interrogation, including information on protein interactions and states, transitions between states and protein function. The expert-authored pages are anonymously peer reviewed by the Nature Publishing Group. The Molecule Pages data is present in an object-relational database format and is freely accessible to the authors, the reviewers and the public from a web browser that serves as a presentation layer. The Molecule Pages are supported by several applications that along with the database and the interfaces form a multi-tier architecture. The Molecule Pages and the Signaling Gateway are routinely accessed by a very large research community. PMID:17965093

  11. The Molecule Pages database.

    PubMed

    Saunders, Brian; Lyon, Stephen; Day, Matthew; Riley, Brenda; Chenette, Emily; Subramaniam, Shankar; Vadivelu, Ilango

    2008-01-01

    The UCSD-Nature Signaling Gateway Molecule Pages (http://www.signaling-gateway.org/molecule) provides essential information on more than 3800 mammalian proteins involved in cellular signaling. The Molecule Pages contain expert-authored and peer-reviewed information based on the published literature, complemented by regularly updated information derived from public data source references and sequence analysis. The expert-authored data includes both a full-text review about the molecule, with citations, and highly structured data for bioinformatics interrogation, including information on protein interactions and states, transitions between states and protein function. The expert-authored pages are anonymously peer reviewed by the Nature Publishing Group. The Molecule Pages data is present in an object-relational database format and is freely accessible to the authors, the reviewers and the public from a web browser that serves as a presentation layer. The Molecule Pages are supported by several applications that along with the database and the interfaces form a multi-tier architecture. The Molecule Pages and the Signaling Gateway are routinely accessed by a very large research community.

  12. Urinary growth hormone level and insulin-like growth factor-1 standard deviation score (IGF-SDS) can discriminate adult patients with severe growth hormone deficiency.

    PubMed

    Hirohata, Toshio; Saito, Nobuhito; Takano, Koji; Yamada, So; Son, Jae-Hyun; Yamada, Shoko M; Nakaguchi, Hiroshi; Hoya, Katsumi; Murakami, Mineko; Mizutani, Akiko; Okinaga, Hiroko; Matsuno, Akira

    2013-01-01

    Adult growth hormone (GH) deficiency (AGHD) in Japan is diagnosed based on peak GH concentrations during GH provocative tests such as GHRP-2 stimulation test. In this study, we aimed to evaluate the ability of serum insulin-like growth factor-1 (sIGF-1) and urinary GH (uGH) at the time of awakening to diagnose AGHD. Fifty-nine patients with pituitary disease (32 men and 27 women; age 20-85 y (57.5 ± 15.5, mean ± SD) underwent GHRP-2 stimulation and sIGF-1 testing. Thirty-six and 23 patients were diagnosed with and without severe AGHD, respectively based on a peak GH response of <9 ng/mL to GHRP-2 stimulation. Serum IGF-1 was evaluated as a standard deviation score (IGF-1 SDS) based on age and sex. We determined whether uGH levels in urine samples from 42 of the 59 patients at awakening were above or below the sensitivity limit. We evaluated IGF-1 SDS and uGH levels in a control group of 15 healthy volunteers. Values for IGF-1 SDS were significantly lower in patients with, than without (-2.07 ± 1.77 vs.-0.03 ± 0.92, mean ± SD; p < 0.001) AGHD whereas the range of IGF-1 SDS substantially overlapped at > -1.4. IGF-1 SDS discriminated AGHD more effectively in patients aged ≤60 years. The χ2 test revealed a statistical relationship between uGH and AGHD (test statistic: 7.0104 ≥ χ2 (1; 0.01) = 6.6349). When IGF-1 SDS is < -1.4 or uGH is below the sensitivity limit, AGHD can be detected with high sensitivity.

  13. Analysis of the SDS-PAGE patterns of outer membrane proteins from Escherichia coli strains that have lost the ability to form K1 antigen and varied in the susceptibility to normal human serum.

    PubMed

    Cisowska, Agnieszka; Bugla-Płoskońska, Gabriela

    2014-01-01

    We used SDS-polyacrylamide gel electrophoresis to investigate the outer membrane proteins (OMPs) band composition of 19 Escherichia coli K1 strains that have spontaneously lost the ability to form K1 polysaccharide capsule (E. coli K1-) and demonstrated different degrees of susceptibility to the bactericidal action of normal human serum. Presented results showed that there were differences between E. coli K1- strains in OMPs expressing capacity. The analysis performed on OMPs has not revealed a direct association between the different OMPs band composition and the susceptibility of these strains to the serum.

  14. Staining Method for Protein Analysis by Capillary Gel Electrophoresis

    PubMed Central

    Wu, Shuqing; Lu, Joann J; Wang, Shili; Peck, Kristy L.; Li, Guigen; Liu, Shaorong

    2009-01-01

    A novel staining method and the associated fluorescent dye were developed for protein analysis by capillary SDS-PAGE. The method strategy is to synthesize a pseudo-SDS dye and use it to replace some of the SDS in SDS–protein complexes so that the protein can be fluorescently detected. The pseudo-SDS dye consists of a long, straight alkyl chain connected to a negative charged fluorescent head and binds to proteins just as SDS. The number of dye molecules incorporated with a protein depends on the dye concentration relative to SDS in the sample solution, since SDS and dye bind to proteins competitively. In this work, we synthesized a series of pseudo-SDS dyes, and tested their performances for capillary SDS-PAGE. FT-16 (a fluorescein molecule linked with a hexadodecyl group) seemed to be the best among all the dyes tested. Although the numbers of dye molecules bound to proteins (and the fluorescence signals from these protein complexes) were maximized in the absence of SDS, high-quality separations were obtained when co-complexes of SDS–protein–dye were formed. The migration time correlates well with protein size even after some of the SDS in the SDS–protein complexes was replaced by the pseudo-SDS dye. Under optimized experimental conditions and using a laser-induced fluorescence detector, limits of detection of as low as 0.13 ng/mL (bovine serum albumin) and dynamic ranges over 5 orders of magnitude in which fluorescence response is proportional to the square root of analyte concentration were obtained. The method and dye were also tested for separations of real-world samples from E. coli. PMID:17874848

  15. Microcephaly Information Page

    MedlinePlus

    ... You are here Home » Disorders » All Disorders Microcephaly Information Page Microcephaly Information Page What research is being done? The National ... the U.S. and Worldwide NINDS Clinical Trials Related Information Patient Organizations Birth Defect Research for Children, Inc. ...

  16. Chapter 07: Species description pages

    Treesearch

    Alex C. Wiedenhoeft

    2011-01-01

    These pages are written to be the final step in the identification process; you will be directed to them by the key in Chapter 6. Each species or group of similar species in the same genus has its own set of pages. The information in the first page describes the characteristics of the wood covered in the manual. The page shows images of similar or confusable woods,...

  17. The Psychometric Properties of the Arabic Version of the Self-Directed Search (SDS)-Form R, 5th Edition for First-Year University Students in the Kingdom of Saudi Arabia (KSA)

    ERIC Educational Resources Information Center

    Akram, Hadeel Abdulah

    2017-01-01

    The factor structure of Holland's hexagonal model as shown in the Self-Directed Search (SDS) has received extensive attention across the world. The goal in creating the SDS was to equip guidance counselors and services with information about adults' personality types, interests, preferences, and career options. More precisely, the SDS items assess…

  18. Investigating the fate of activated sludge extracellular proteins in sludge digestion using sodium dodecyl sulfate polyacrylamide gel electrophoresis.

    PubMed

    Park, Chul; Helm, Richard F; Novak, John T

    2008-12-01

    The fate of activated sludge extracellular proteins in sludge digestion was investigated using three different cation-associated extraction methods and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Extraction methods used were the cation exchange resin (CER) method for extracting calcium (Ca2+) and magnesium (Mg2+), sulfide extraction for removing iron, and base treatment (pH 10.5) for dissolving aluminum. Extracellular polymeric substances extracted were then subjected to SDS-PAGE, and the resultant protein profiles were examined before and after sludge digestion. The SDS-PAGE results showed that three methods led to different SDS-PAGE profiles for both undigested and digested sludges. The results further revealed that CER-extracted proteins remained mainly undegraded in anaerobic digestion, but were degraded in aerobic digestion. While the fate of sulfide- and base-extracted proteins was not clear for aerobic digestion, their changes in anaerobic digestion were elucidated. Most sulfide-extracted proteins were removed by anaerobic digestion, while the increase in protein band intensity and diversity was observed for base-extracted proteins. These results suggest that activated sludge flocs contain different fractions of proteins that are distinguishable by their association with certain cations and that each fraction undergoes different fates in anaerobic and aerobic digestion. The proteins that were resistant to degradation and generated during anaerobic digestion were identified by liquid chromatography tandem mass spectrometry. Protein identification results and their putative roles in activated sludge and anaerobic digestion are discussed in this study.

  19. Investigation the foam dynamics capacity of SDS in foam generator by affecting the presence of organic and inorganic contaminant

    NASA Astrophysics Data System (ADS)

    Haryanto, Bode; Siswarni, M. Z.; Sianipar, Yosef C. H.; Sinaga, Tongam M. A.; Bestari, Imam

    2017-05-01

    The effect of negative charge SDS monomer on its foam capacity with the presence of contaminants was investigated in foam generator. Generally, surfactant with higher concentration has higher foam capacity. The higher concentration will increase the number of monomer then increase the micelles in liquid phase. Increasing the number of monomer with the negative charge is a potential to increase interaction with metal ion with positive charge in solution. The presence of inorganic compound as metal ion with positive charge and organic compound (colloid) as particle of coffee impacting to generate the foam lamella with monomer is evaluated. Foam dynamic capacity of only SDS with variation of CMC, 1 x; 2 x; 3 x have the height 7.5, 8.0 and 8.3 cm respectively with the different range time were investigated. The Height of foam dynamic capacity with the presence of 20 ppm Cd2+ ion contaminant was 8.0, 8.3 and 8.4 cm at the same CMC variation of SDS. The presence of metal ion contaminant within the foam was confirmed by AAS. The black coffee particles and oil as contaminant decreased the foam capacity significantly in comparing to metal ions.

  20. Coarse-Grain Molecular Dynamics Simulations To Investigate the Bulk Viscosity and Critical Micelle Concentration of the Ionic Surfactant Sodium Dodecyl Sulfate (SDS) in Aqueous Solution.

    PubMed

    Ruiz-Morales, Yosadara; Romero-Martínez, Ascención

    2018-04-12

    The first critical micelle concentration (CMC) of the ionic surfactant sodium dodecyl sulfate (SDS) in diluted aqueous solution has been determined at room temperature from the investigation of the bulk viscosity, at several concentrations of SDS, by means of coarse-grain molecular dynamics simulations. The coarse-grained model molecules at the mesoscale level are adopted. The bulk viscosity of SDS was calculated at several millimolar concentrations of SDS in water using the MARTINI force field by means of NVT shear Mesocite molecular dynamics. The definition of each bead in the MARTINI force field is established, as well as their radius, volume, and mass. The effect of the size of the simulation box on the obtained CMC has been investigated, as well as the effect of the number of SDS molecules, in the simulations, on the formation of aggregates. The CMC, which was obtained from a graph of the calculated viscosities versus concentration, is in good agreement with the reported experimental data and does not depend on the size of the box used in the simulation. The formation of a spherical micelle-like aggregate is observed, where the dodecyl sulfate tails point inward and the heads point outward the aggregation micelle, in accordance with experimental observations. The advantage of using coarse-grain molecular dynamics is the possibility of treating explicitly charged beads, applying a shear flow for viscosity calculation, and processing much larger spatial and temporal scales than atomistic molecular dynamics can. Furthermore, the CMC of SDS obtained with the coarse-grained model is in much better agreement with the experimental value than the value obtained with atomistic simulations.

  1. Realistic page-turning of electronic books

    NASA Astrophysics Data System (ADS)

    Fan, Chaoran; Li, Haisheng; Bai, Yannan

    2014-01-01

    The booming electronic books (e-books), as an extension to the paper book, are popular with readers. Recently, many efforts are put into the realistic page-turning simulation o f e-book to improve its reading experience. This paper presents a new 3D page-turning simulation approach, which employs piecewise time-dependent cylindrical surfaces to describe the turning page and constructs smooth transition method between time-dependent cylinders. The page-turning animation is produced by sequentially mapping the turning page into the cylinders with different radii and positions. Compared to the previous approaches, our method is able to imitate various effects efficiently and obtains more natural animation of turning page.

  2. The Faculty Web Page: Contrivance or Continuation?

    ERIC Educational Resources Information Center

    Lennex, Lesia

    2007-01-01

    In an age of Internet education, what does it mean for a tenure/tenure-track faculty to have a web page? How many professors have web pages? If they have a page, what does it look like? Do they really need a web page at all? Many universities have faculty web pages. What do those collective pages look like? In what way do they represent the…

  3. Reese Sorenson's Individual Professional Page

    NASA Technical Reports Server (NTRS)

    Sorenson, Reese; Nixon, David (Technical Monitor)

    1998-01-01

    The subject document is a World Wide Web (WWW) page entitled, "Reese Sorenson's Individual Professional Page." Its can be accessed at "http://george.arc.nasa.gov/sorenson/personal/index.html". The purpose of this page is to make the reader aware of me, who I am, and what I do. It lists my work assignments, my computer experience, my place in the NASA hierarchy, publications by me, awards received by me, my education, and how to contact me. Writing this page was a learning experience, pursuant to an element in my Job Description which calls for me to be able to use the latest computers. This web page contains very little technical information, none of which is classified or sensitive.

  4. Soybean SDS in South Africa is caused by Fusarium brasiliense and a novel undescribed Fusarium sp.

    USDA-ARS?s Scientific Manuscript database

    Soybean sudden death syndrome (SDS) was detected in South Africa for the first time during pathogen surveys conducted in 2013-2014. The primary objective of this study was to characterize the 16 slow-growing Fusarium strains that were isolated from the roots of symptomatic plants. Molecular phylogen...

  5. The NPOESS Preparatory Project (NPP) Science Data Segment (SDS) Data Depository and Distribution Element (SD3E) System Architecture

    NASA Technical Reports Server (NTRS)

    Ho, Evelyn L.; Schweiss, Robert J.

    2008-01-01

    The National Polar-orbiting Operational Environmental Satellite System (NPOESS), the U.S. Government's future low-Earth orbiting satellite system, will monitor global weather and environmental conditions. Serving as a risk reduction for NPOESS, the NPOESS Preparatory Project (NPP) will provide remotely sensed atmospheric, land, ocean, ozone, and sounder data that will serve the meteorological and global climate change scientific communities. The National Aeronautics and Space Administration (NASA) NPP Science Data Segment's (SDS) primary role is to independently assess the quality of the NPP science and environmental data records for their ability to support climate research. The SDS is composed of nine elements; an input element that receives data from the operational agencies and acts as a buffer, a calibration analysis element, five elements devoted to measurement based quality assessment, an element used to test algorithmic improvements, and an element that provides overall science direction. Each element requires a set of sensor specific science data products for their evaluation. There are four NPP sensors that will be flown on the NPP observatory. They are the Visible Infrared Imagining Radiometer Suite (VIIRS), the Advanced Technology Microwave Sounder (ATMS), the Cross-Track Infrared Sounder (CrIS), and the Ozone Mapper/Profiler Suite (OMPS). It is estimated that these four sensors combined will make daily data requests for approximately six terabytes of NPP science products from the operational data providers. As a result, issues associated with duplicate data requests, data transfers of large volumes of diverse products, and data transfer failures raised concerns with respect to the network traffic and bandwidth consumption. Therefore, a central data broker system for receiving and buffering data requests and data products for the SDS was developed. The data element for this system is called the SDS Data Depository and Distribution Element (SD3E). It

  6. [Effect of gene optimization on the expression and purification of HDV small antigen produced by genetic engineering].

    PubMed

    Ding, Jun-Ying; Meng, Qing-Ling; Guo, Min-Zhuo; Yi, Yao; Su, Qiu-Dong; Lu, Xue-Xin; Qiu, Feng; Bi, Sheng-Li

    2012-10-01

    To study the effect of gene optimization on the expression and purification of HDV small antigen produced by genetic engineering. Based on the colon preference of E. coli, the HDV small antigen original gene from GenBank was optimized. Both the original gene and the optimized gene expressed in prokaryotic cells, SDS-PAGE was made to analyze the protein expression yield and to decide which protein expression style was more proportion than the other. Furthermore, two antigens were purified by chromatography in order to compare the purity by SDS-PAGE and Image Lab software. SDS-PAGE indicated that the molecular weight of target proteins from two groups were the same as we expected. Gene optimization resulted in the higher yield and it could make the product more soluble. After chromatography, the purity of target protein from optimized gene was up to 96.3%, obviously purer than that from original gene. Gene optimization could increase the protein expression yield and solubility of genetic engineering HDV small antigen. In addition, the product from the optimized gene group was easier to be purified for diagnosis usage.

  7. Full Page Departmental Advertising.

    ERIC Educational Resources Information Center

    Van Zante, Ben

    1978-01-01

    States that many school newspapers are condensing all advertising into one or two pages. Indicates that advertisers find this to be acceptable, students continue to read the ads, and the content pages look better. (TJ)

  8. Structural characteristics of Tla products

    PubMed Central

    1985-01-01

    Biochemical study of thymus leukemia antigen (TL) from thymocytes of various Tla genotypes and from leukemia cells revealed features that, given present evidence, are peculiar to TL among class I products of the H-2:Qa:Tla region of chromosome 17. Sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) of TL from thymocytes of all TL+ mouse strains, precipitated by anti-TL antiserum or monoclonal antibodies, showed two closely migrating bands of equal intensity in the heavy (H) chain position (45-50,000 mol wt). Comparison of these two bands by two-dimensional isoelectric focusing (2D IEF)-SDS-PAGE and 2D chymotryptic peptide mapping showed no differences indicative of protein dissimilarity. Thus, the two components of the H chain doublet may differ only in a feature of glycosylation that does not affect charge. The two leukemias studied gave only a single band in the H chain position. On 2D peptide mapping and 2D IEF-SDS-PAGE, the patterns for TL of Tlaa and Tlae thymocytes, which are closely related serologically, were broadly similar, but clearly different from the pattern typical of Tlac and Tlad thymocytes. 2D peptide maps of TL from Tlaa thymocytes and Tlaa leukemia cells did not differ. Leukemia cells of Tlab origin (thymocytes TL-) gave 2D peptide and 2D IEF-SDS-PAGE patterns of a third type. With the exception of Tlaa, thymocytes of TL+ mice yielded additional TL products of higher molecular weight than the TL H chain. PMID:3875681

  9. Frequently Asked Questions | DOE PAGES

    Science.gov Websites

    read, download, and analyze at no charge to users. DOE PAGES offers free public access to the best manuscripts are hosted, DOE PAGES enables readers to search them all via a single query. In most cases, free . DOE PAGES offers free public access to the best available full-text version of DOE-funded scholarly

  10. Sleep Apnea Information Page

    MedlinePlus

    ... are here Home » Disorders » All Disorders Sleep Apnea Information Page Sleep Apnea Information Page What research is being done? The National ... the U.S. and Worldwide NINDS Clinical Trials Related Information Patient Organizations American Sleep Apnea Association American Sleep ...

  11. Sign language Web pages.

    PubMed

    Fels, Deborah I; Richards, Jan; Hardman, Jim; Lee, Daniel G

    2006-01-01

    The WORLD WIDE WEB has changed the way people interact. It has also become an important equalizer of information access for many social sectors. However, for many people, including some sign language users, Web accessing can be difficult. For some, it not only presents another barrier to overcome but has left them without cultural equality. The present article describes a system that allows sign language-only Web pages to be created and linked through a video-based technique called sign-linking. In two studies, 14 Deaf participants examined two iterations of signlinked Web pages to gauge the usability and learnability of a signing Web page interface. The first study indicated that signing Web pages were usable by sign language users but that some interface features required improvement. The second study showed increased usability for those features; users consequently couldnavigate sign language information with ease and pleasure.

  12. Sjogren's Syndrome Information Page

    MedlinePlus

    ... are here Home » Disorders » All Disorders Sjögren's Syndrome Information Page Sjögren's Syndrome Information Page What research is being done? The goals ... the U.S. and Worldwide NINDS Clinical Trials Related Information Patient Organizations Arthritis Foundation National Eye Institute (NEI) ...

  13. Insights into Facebook Pages: an early adolescent health research study page targeted at parents.

    PubMed

    Amon, Krestina L; Paxton, Karen; Klineberg, Emily; Riley, Lisa; Hawke, Catherine; Steinbeck, Katharine

    2016-02-01

    Facebook has been used in health research, but there is a lack of literature regarding how Facebook may be used to recruit younger adolescents. A Facebook Page was created for an adolescent cohort study on the effects of puberty hormones on well-being and behaviour in early adolescence. Used as a communication tool with existing participants, it also aimed to alert potential participants to the study. The purpose of this paper is to provide a detailed description of the development of the study Facebook Page and present the fan response to the types of posts made on the Page using the Facebook-generated Insights data. Two types of posts were made on the study Facebook Page. The first type was study-related update posts and events. The second was relevant adolescent and family research and current news posts. Observations on the use of and response to the Page were made over 1 year across three phases (phase 1, very low Facebook use; phase 2, high Facebook use; phase 3, low Facebook use). Most Page fans were female (88.6%), with the largest group of fans aged between 35 and 44 years. Study-related update posts with photographs were the most popular. This paper provides a model on which other researchers could base Facebook communication and potential recruitment in the absence of established guidelines.

  14. Proteomics analysis in frozen horse mackerel previously high-pressure processed.

    PubMed

    Pazos, Manuel; Méndez, Lucía; Vázquez, Manuel; Aubourg, Santiago P

    2015-10-15

    The effect of high-pressure processing (HPP) (150, 300 and 450 MPa for 0, 2.5 and 5 min) on total sodium dodecyl sulphate (SDS)-soluble and sarcoplasmic proteins in frozen (-10 °C for 3 months) horse mackerel (Trachurus trachurus) was evaluated. Proteomics tools based on image analysis of SDS-PAGE protein gels and protein identification by tandem mass spectrometry (MS/MS) were applied. Although total SDS-soluble fraction indicated no important changes induced by HPP, this processing modified the 1-D SDS-PAGE sarcoplasmic patterns in a direct-dependent manner and exerted a selective effect on particular proteins depending on processing conditions. Thus, application of the highest pressure (450 MPa) provoked a significant degradation of phosphoglycerate mutase 2, glycogen phosphorylase muscle form, pyruvate kinase muscle isozyme, beta-enolase and triosephosphate isomerase and phosphoglucomutase-1. Conversely, protein bands assigned to tropomyosin alpha-1 chain, fast myotomal muscle troponin T and parvalbumin beta 2 increased their intensity after applying a 450-MPa processing. Copyright © 2015 Elsevier Ltd. All rights reserved.

  15. Finding Specification Pages from the Web

    NASA Astrophysics Data System (ADS)

    Yoshinaga, Naoki; Torisawa, Kentaro

    This paper presents a method of finding a specification page on the Web for a given object (e.g., ``Ch. d'Yquem'') and its class label (e.g., ``wine''). A specification page for an object is a Web page which gives concise attribute-value information about the object (e.g., ``county''-``Sauternes'') in well formatted structures. A simple unsupervised method using layout and symbolic decoration cues was applied to a large number of the Web pages to acquire candidate attributes for each class (e.g., ``county'' for a class ``wine''). We then filter out irrelevant words from the putative attributes through an author-aware scoring function that we called site frequency. We used the acquired attributes to select a representative specification page for a given object from the Web pages retrieved by a normal search engine. Experimental results revealed that our system greatly outperformed the normal search engine in terms of this specification retrieval.

  16. Envelope protein complexes of Mycobacterium avium subsp. paratuberculosis and their antigenicity

    USDA-ARS?s Scientific Manuscript database

    Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of Johne’s disease, a chronic enteric disease of ruminant animals. In the present study, blue native PAGE electrophoresis and 2D SDS-PAGE were used to separate MAP envelope protein complexes, followed by mass spectrometry (MS) ...

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  1. High-molecular weight Aβ oligomers and protofibrils are the predominant Aβ species in the native soluble protein fraction of the AD brain.

    PubMed

    Upadhaya, Ajeet Rijal; Lungrin, Irina; Yamaguchi, Haruyasu; Fändrich, Marcus; Thal, Dietmar Rudolf

    2012-02-01

    Alzheimer's disease (AD) is characterized by the aggregation and deposition of amyloid β protein (Aβ) in the brain. Soluble Aβ oligomers are thought to be toxic. To investigate the predominant species of Aβ protein that may play a role in AD pathogenesis, we performed biochemical analysis of AD and control brains. Sucrose buffer-soluble brain lysates were characterized in native form using blue native (BN)-PAGE and also in denatured form using SDS-PAGE followed by Western blot analysis. BN-PAGE analysis revealed a high-molecular weight smear (>1000 kD) of Aβ(42) -positive material in the AD brain, whereas low-molecular weight and monomeric Aβ species were not detected. SDS-PAGE analysis, on the other hand, allowed the detection of prominent Aβ monomer and dimer bands in AD cases but not in controls. Immunoelectron microscopy of immunoprecipitated oligomers and protofibrils/fibrils showed spherical and protofibrillar Aβ-positive material, thereby confirming the presence of high-molecular weight Aβ (hiMWAβ) aggregates in the AD brain. In vitro analysis of synthetic Aβ(40) - and Aβ(42) preparations revealed Aβ fibrils, protofibrils, and hiMWAβ oligomers that were detectable at the electron microscopic level and after BN-PAGE. Further, BN-PAGE analysis exhibited a monomer band and less prominent low-molecular weight Aβ (loMWAβ) oligomers. In contrast, SDS-PAGE showed large amounts of loMWAβ but no hiMWAβ(40) and strikingly reduced levels of hiMWAβ(42) . These results indicate that hiMWAβ aggregates, particularly Aβ(42) species, are most prevalent in the soluble fraction of the AD brain. Thus, soluble hiMWAβ aggregates may play an important role in the pathogenesis of AD either independently or as a reservoir for release of loMWAβ oligomers. © 2011 The Authors Journal of Cellular and Molecular Medicine © 2011 Foundation for Cellular and Molecular Medicine/Blackwell Publishing Ltd.

  2. High-molecular weight Aβ oligomers and protofibrils are the predominant Aβ species in the native soluble protein fraction of the AD brain

    PubMed Central

    Upadhaya, Ajeet Rijal; Lungrin, Irina; Yamaguchi, Haruyasu; Fändrich, Marcus; Thal, Dietmar Rudolf

    2012-01-01

    Abstract Alzheimer’s disease (AD) is characterized by the aggregation and deposition of amyloid β protein (Aβ) in the brain. Soluble Aβ oligomers are thought to be toxic. To investigate the predominant species of Aβ protein that may play a role in AD pathogenesis, we performed biochemical analysis of AD and control brains. Sucrose buffer-soluble brain lysates were characterized in native form using blue native (BN)-PAGE and also in denatured form using SDS-PAGE followed by Western blot analysis. BN-PAGE analysis revealed a high-molecular weight smear (>1000 kD) of Aβ42-positive material in the AD brain, whereas low-molecular weight and monomeric Aβ species were not detected. SDS-PAGE analysis, on the other hand, allowed the detection of prominent Aβ monomer and dimer bands in AD cases but not in controls. Immunoelectron microscopy of immunoprecipitated oligomers and protofibrils/fibrils showed spherical and protofibrillar Aβ-positive material, thereby confirming the presence of high-molecular weight Aβ (hiMWAβ) aggregates in the AD brain. In vitro analysis of synthetic Aβ40- and Aβ42 preparations revealed Aβ fibrils, protofibrils, and hiMWAβ oligomers that were detectable at the electron microscopic level and after BN-PAGE. Further, BN-PAGE analysis exhibited a monomer band and less prominent low-molecular weight Aβ (loMWAβ) oligomers. In contrast, SDS-PAGE showed large amounts of loMWAβ but no hiMWAβ40 and strikingly reduced levels of hiMWAβ42. These results indicate that hiMWAβ aggregates, particularly Aβ42 species, are most prevalent in the soluble fraction of the AD brain. Thus, soluble hiMWAβ aggregates may play an important role in the pathogenesis of AD either independently or as a reservoir for release of loMWAβ oligomers. PMID:21418518

  3. Identification of Psilocybe cubensis spore allergens by immunoprinting.

    PubMed

    Helbling, A; Horner, W E; Lehrer, S B

    1993-01-01

    Previous studies established that Psilocybe cubensis contains potent allergens, and that a significant percentage of atopic subjects were sensitized to P. cubensis spores. The objective of this study was to identify P. cubensis spore allergens using isoelectric focusing (IEF) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) immunoprinting. Coomassie blue staining of IEF gels detected approximately 20 bands between pI 3.6 and 9.3. Immunoprints obtained with 15 P. cubensis skin test- and RAST-positive sera revealed 13 IgE-binding bands; the most reactive were at pI 5.0 (80%), 5.6 (87%), 8.7 (80%) and 9.3 (100%). SDS-PAGE resolved 27 proteins ranging from about 13 to 112 kD. SDS-PAGE immunoprints conducted with 11 skin test- and RAST-positive sera demonstrated 18 IgE-binding bands; most sera reacted to 16 (82%), 35 (100%) and 76 kD (91%) allergens. Both electrophoretic procedures demonstrated a single allergen (at pI 9.3 and 35 kD) that reacted with all sera tested. This study corroborates the allergenic significance of P. cubensis spores and identifies the allergens of greatest importance.

  4. Cross-reactivity between pollen extracts from six artemisia species.

    PubMed

    Brandys, J; Grimsøen, A; Nilsen, B M; Paulsen, B S; Park, H S; Hong, C S

    1993-06-01

    Pollen extracts of six different ARTEMISIA species, A. VULGARIS, A. SCOPARIA, A. PRINCEPS, A. TRIDENTATA, A. ANNUA, and A. CAMPESTRIS were compared using SDS-PAGE, IEF, immunoblotting, and immunoelectrophoretic methods. The band patterns obtained after SDS-PAGE and IEF showed a large degree of similarity between the extracts. Immunoblotting of these gels using a pool of sera from patients allergic to A. VULGARIS gave essentially the same IgE-binding band pattern with all the extracts, demonstrating an extensive degree of cross-reactivity between A. VULGARIS and the other ARTEMISIA species. FRIE using a polyspecific antiserum against A. VULGARIS showed that all the extracts contained several antigens that were immunologically identical to antigens in A. VULGARIS extract. Antigens showing immunological identity to the important A. VULGARIS allergens Ag 12 and ART V II were present in all the extracts. The cross-reactivity between A. VULGARIS and A. PRINCEPS was further verified by screening of ten Korean and nine Norwegian individual patient sera against extracts of both species in SDS-PAGE or IEF immunoblotting. Both groups of patients had essentially the same pattern of reactivity towards both pollen extracts.

  5. Purification and characterization of selenium-containing phycocyanin from selenium-enriched Spirulina platensis.

    PubMed

    Chen, Tianfeng; Wong, Yum-Shing; Zheng, Wenjie

    2006-11-01

    A fast protein liquid chromatographic method for purification of selenium-containing phycocyanin (Se-PC) from selenium-enriched Spirulina platensis was described in this study. The purification procedures involved fractionation by ammonium sulfate precipitation, DEAE-Sepharose ion-exchange chromatography and Sephacry S-300 size exclusion chromatography. The purity ratio (A620/A280) and the separation factor (A620/A655) of the purified Se-PC were 5.12 and 7.92, respectively. The Se concentration of purified Se-PC was 496.5 microg g(-1) protein, as determined by ICP-AES analysis. The purity of the Se-PC was further characterized by UV-VIS and fluorescence spectrometry, SDS-PAGE, RP-HPLC and gel filtration HPLC. The apparent molecular mass of the native Se-PC determined by gel filtration HPLC was 109 kDa, indicating that the protein existed as a trimer. SDS-PAGE of the purified Se-PC yielded two major bands corresponding to the alpha and beta subunits. A better separation of these two subunits was obtained by RP-HPLC. Identification of the alpha and beta subunits separated by SDS-PAGE and RP-HPLC was achieved by peptide mass fingerprinting (PMF) using MALDI-TOF-TOF mass spectrometry.

  6. About / FAQ | DOE PAGES

    Science.gov Websites

    : + Advanced Search × Advanced Search All Fields: Title: Full Text: Bibliographic Data: Creator / Author: Name discoverable at no charge to users. DOE PAGES offers free public access to the best available full-text version and distributed content, with PAGES maintaining a permanent archive of all full text and metadata. In

  7. Conditions that allow for effective transfer of membrane proteins onto nitrocellulose membrane in Western blots.

    PubMed

    Abeyrathne, Priyanka D; Lam, Joseph S

    2007-04-01

    A major hurdle in characterizing bacterial membrane proteins by Western blotting is the ineffectiveness of transferring these proteins from sodium dodecyl sulfate -- polyacrylamide gel electrophoresis (SDS-PAGE) gel onto nitrocellulose membrane, using standard Western blot buffers and electrophoretic conditions. In this study, we compared a number of modified Western blotting buffers and arrived at a composition designated as the SDS-PAGE-Urea Lysis buffer. The use of this buffer and specific conditions allowed the reproducible transfer of highly hydrophobic bacterial membrane proteins with 2-12 transmembrane-spanning segments as well as soluble proteins onto nitrocellulose membranes. This method should be broadly applicable for immunochemical studies of other membrane proteins.

  8. Visualizing Rank Time Series of Wikipedia Top-Viewed Pages.

    PubMed

    Xia, Jing; Hou, Yumeng; Chen, Yingjie Victor; Qian, Zhenyu Cheryl; Ebert, David S; Chen, Wei

    2017-01-01

    Visual clutter is a common challenge when visualizing large rank time series data. WikiTopReader, a reader of Wikipedia page rank, lets users explore connections among top-viewed pages by connecting page-rank behaviors with page-link relations. Such a combination enhances the unweighted Wikipedia page-link network and focuses attention on the page of interest. A set of user evaluations shows that the system effectively represents evolving ranking patterns and page-wise correlation.

  9. Protein extraction into the bicontinuous microemulsion phase of a Water/SDS/pentanol/dodecane winsor-III system: Effect on nanostructure and protein conformation

    DOE PAGES

    Hayes, Douglas G.; Ye, Ran; Dunlap, Rachel N.; ...

    2017-09-07

    Bicontinuous microemulsions (BμEs), consisting of water and oil nanodomains separated by surfactant monolayers of near-zero curvature, are potentially valuable systems for purification and delivery of biomolecules, for hosting multiphasic biochemical reactions, and as templating media for preparing nanomaterials. We formed Winsor-III systems by mixing aqueous protein and sodium dodecyl sulfate (SDS) solutions with dodecane and 1-pentanol (cosurfactant) to efficiently extract proteins into the middle (BμE) phase. Bovine serum albumin (BSA) and cytochrome c partitioned to the BμE phase at 64% and 81% efficiency, respectively, producing highly concentrated protein solutions (32 and 44 g L –1, respectively), through release of watermore » and oil from the BμEs. Circular dichroism spectroscopic analysis demonstrated that BSA underwent minor secondary structural changes upon incorporation into BμEs, while the secondary structure of cytochrome c and pepsin underwent major changes. Small-angle x-ray scattering (SAXS) results show that proteins promoted an increase of the interfacial fluidity and surface area per volume for the BμE surfactant monolayers, and that each protein uniquely altered self-assembly in the Winsor-III systems. Cytochrome c partitioned via electrostatic attractions between SDS and the protein’s positively-charged groups, residing near the surfactant head groups of BμE monolayers, where it decreased surfactant packing efficiency. BSA partitioned through formation of SDS-BSA complexes via hydrophobic and electrostatic attractive interactions. As the BSA-SDS ratio increased, complexes’ partitioning favored BμEs over the oil excess phase due to the increased hydrophilicity of the complexes. In conclusion, this study demonstrates the potential utility of BμEs to purify proteins and prepare nanostructured fluids possessing high protein concentration.« less

  10. Protein extraction into the bicontinuous microemulsion phase of a Water/SDS/pentanol/dodecane winsor-III system: Effect on nanostructure and protein conformation

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Hayes, Douglas G.; Ye, Ran; Dunlap, Rachel N.

    Bicontinuous microemulsions (BμEs), consisting of water and oil nanodomains separated by surfactant monolayers of near-zero curvature, are potentially valuable systems for purification and delivery of biomolecules, for hosting multiphasic biochemical reactions, and as templating media for preparing nanomaterials. We formed Winsor-III systems by mixing aqueous protein and sodium dodecyl sulfate (SDS) solutions with dodecane and 1-pentanol (cosurfactant) to efficiently extract proteins into the middle (BμE) phase. Bovine serum albumin (BSA) and cytochrome c partitioned to the BμE phase at 64% and 81% efficiency, respectively, producing highly concentrated protein solutions (32 and 44 g L –1, respectively), through release of watermore » and oil from the BμEs. Circular dichroism spectroscopic analysis demonstrated that BSA underwent minor secondary structural changes upon incorporation into BμEs, while the secondary structure of cytochrome c and pepsin underwent major changes. Small-angle x-ray scattering (SAXS) results show that proteins promoted an increase of the interfacial fluidity and surface area per volume for the BμE surfactant monolayers, and that each protein uniquely altered self-assembly in the Winsor-III systems. Cytochrome c partitioned via electrostatic attractions between SDS and the protein’s positively-charged groups, residing near the surfactant head groups of BμE monolayers, where it decreased surfactant packing efficiency. BSA partitioned through formation of SDS-BSA complexes via hydrophobic and electrostatic attractive interactions. As the BSA-SDS ratio increased, complexes’ partitioning favored BμEs over the oil excess phase due to the increased hydrophilicity of the complexes. In conclusion, this study demonstrates the potential utility of BμEs to purify proteins and prepare nanostructured fluids possessing high protein concentration.« less

  11. NMR structure of the Arctic mutation of the Alzheimer's Aβ(1-40) peptide docked to SDS micelles

    NASA Astrophysics Data System (ADS)

    Usachev, K. S.; Filippov, A. V.; Khairutdinov, B. I.; Antzutkin, O. N.; Klochkov, V. V.

    2014-11-01

    The “Arctic” point mutation of the Alzheimer's amyloid β-peptide is a rare mutation leading to an early onset of Alzheimer's disease. The peptide may interact with neuronal membranes, where it can provide its toxic effects. We used 2D NMR spectroscopy to investigate the conformation of the “Arctic” mutant of Aβ1-40 Alzheimer's amyloid peptide in sodium dodecyl sulfate micelle solutions, which are the type of amphiphilic structures mimicking some properties of biomembranes. The study showed that the Arctic mutant of Aβ1-40 interacts with the surface of SDS micelles mainly through the Leu17-Asn27 310-helical region, while the Ile31-Val40 region is buried in the hydrophobic interior of the micelle. In contrast, wild-type Aβ1-40 interacts with SDS micelles through the Lys16-Asp23 α-helical region and Gly29-Met35. Both the Arctic mutant and the wild-type Aβ1-40 peptides interactions with SDS micelles are hydrophobic in nature. Aβ peptides are thought to be capable of forming pores in biomembranes that can cause changes in neuronal and endothelial cell membrane permeability. It has also been shown that Aβ peptides containing the “Arctic” mutation are more neurotoxic and aggregate more readily than the wild-type Aβ peptides at physiological conditions. Here, we propose that the extension of the helical structure of Leu17-Asn27 and a high aliphaticity (neutrality) of the C-terminal region in the Arctic Aβ peptides are consistent with the idea that formation of ion-permeable pores by Aβ oligomers may be one of prevailing mechanisms of a larger neuronal toxicity of the Arctic Aβ compared to the wild-type Aβ peptides, independent of oxidative damage and lipid peroxidation.

  12. Delivery of RNA and its intracellular translation into protein mediated by SDS-CTAB vesicles: potential use in nanobiotechnology.

    PubMed

    Russo, Laura; Berardi, Valerio; Tardani, Franco; La Mesa, Camillo; Risuleo, Gianfranco

    2013-01-01

    Catanionic vesicles are supramolecular aggregates spontaneously forming in water by electrostatic attraction between two surfactants mixed in nonstoichiometric ratios. The outer surface charges allow adsorption to the biomembrane by electrostatic interactions. The lipoplex thus obtained penetrates the cell by endocytosis or membrane fusion. We examined the possible cytotoxic effects and evaluated the transfection efficiency of one vesicle type as compared to known commercial carriers. We show that the individual components of two different vesicles types, CTAB (cetyltrimethylammonium bromide) and DDAB (didodecyldimethylammonium bromide) are detrimental for cell survival. We also assayed the cytotoxicity of SDS-DDAB vesicles and showed dose and time dependency, with the DDAB component being per se extremely cytotoxic. The transfection efficiency of exogenous RNA mediated by SDS-CTAB increases if vesicles assemble in the presence of the reporter RNA; finally, freezing abrogates the transfection ability. The results of our experimental strategy suggest that catanionic vesicles may be adopted in gene therapy and control of antiproliferative diseases.

  13. 14 CFR 221.31 - Title page.

    Code of Federal Regulations, 2013 CFR

    1998-01-01

    ... 14 Aeronautics and Space 4 1998-01-01 1998-01-01 false Title page. 221.31 Section 221.31 ECONOMIC REGULATIONS TARIFFS Contents of Tariff § 221.31 Title page. (a) Contents. Except as otherwise required in this part, or by other regulatory agencies, the title page of every tariff shall contain the following information to be shown in the order...

  14. 14 CFR 221.31 - Title page.

    Code of Federal Regulations, 2011 CFR

    1999-01-01

    ... 14 Aeronautics and Space 4 1999-01-01 1999-01-01 false Title page. 221.31 Section 221.31 ECONOMIC REGULATIONS TARIFFS Contents of Tariff § 221.31 Title page. (a) Contents. Except as otherwise required in this part, or by other regulatory agencies, the title page of every tariff shall contain the following information to be shown in the order...

  15. 14 CFR 221.31 - Title page.

    Code of Federal Regulations, 2012 CFR

    1997-01-01

    ... 14 Aeronautics and Space 4 1997-01-01 1997-01-01 false Title page. 221.31 Section 221.31 ECONOMIC REGULATIONS TARIFFS Contents of Tariff § 221.31 Title page. (a) Contents. Except as otherwise required in this part, or by other regulatory agencies, the title page of every tariff shall contain the following information to be shown in the order...

  16. Antihemorrhagin in the blood serum of king cobra (Ophiophagus hannah): purification and characterization.

    PubMed

    Chanhome, Lawan; Khow, Orawan; Omori-Satoh, Tamotsu; Sitprija, Visith

    2003-06-01

    King cobra (Ophiophagus hannah) serum was found to possess antihemorrhagic activity against king cobra hemorrhagin. The activity was stronger than that in commercial king cobra antivenom. An antihemorrhagin has been purified by ion exchange chromatography, affinity chromatography and gel filtration with a 22-fold purification and an overall yield of 12% of the total antihemorrhagic activity contained in crude serum. The purified antihemorrhagin was homogeneous in disc-PAGE and SDS-PAGE. Its apparent molecular weight determined by SDS-PAGE was 120 kDa. The antihemorrhagin was also active against other hemorrhagic snake venoms obtained in Thailand and Japan such as Calloselasma rhodostoma, Trimeresurus albolabris, Trimeresurus macrops and Trimeresurus flavoviridis (Japanese Habu). It inhibited the proteolytic activity of king cobra venom. It is an acid- and thermolabile protein and does not form precipitin lines against king cobra venom.

  17. Fluorescence and electron paramagnetic resonance studies of norfloxacin and N-donor mixed-ligand ternary copper(II) complexes: Stability and interaction with SDS micelles

    NASA Astrophysics Data System (ADS)

    Vignoli Muniz, Gabriel S.; Incio, Jimmy Llontop; Alves, Odivaldo C.; Krambrock, Klaus; Teixeira, Letícia R.; Louro, Sonia R. W.

    2018-01-01

    The stability of ternary copper(II) complexes of a heterocyclic ligand, L (L being 2,2‧-bipyridine (bipy) or 1,10-phenanthroline (phen)) and the fluorescent antibacterial agent norfloxacin (NFX) as the second ligand was studied at pH 7.4 and different ionic strengths. Fluorescence quenching upon titration of NFX with the binary complexes allowed to obtain stability constants for NFX binding, Kb, as a function of ionic strength. The Kb values vary by more than two orders of magnitude when buffer concentration varies from 0.5 to 100 mM. It was observed that previously synthesized ternary complexes dissociate in buffer according with the obtained stability constants. This shows that equimolar solutions of NFX and binary complexes are equivalent to solutions of synthesized ternary complexes. The interaction of the ternary copper complexes with anionic SDS (sodium dodecyl sulfate) micelles was studied by fluorescence and electron paramagnetic resonance (EPR). Titration of NFX-loaded SDS micelles with the complexes Cu:L allowed to determine the stability constants inside the micelles. Fluorescence quenching demonstrated that SDS micelles increase the stability constants by factors around 50. EPR spectra gave details of the copper(II) local environment, and demonstrated that the structure of the ternary complexes inside SDS micelles is different from that in buffer. Mononuclear ternary complexes formed inside the micelles, while in buffer most ternary complexes are binuclear. The results show that anionic membrane interfaces increase formation of copper fluoroquinolone complexes, which can influence bioavailability, membrane diffusion, and mechanism of action of the antibiotics.

  18. Concepciones y concepciones alternativas de estudiantes universitarios/as de biologia y futuros maestros/as de Ciencia de escuela secundaria sobre la teoria de evolucion biologica por seleccion natural

    NASA Astrophysics Data System (ADS)

    Morales Ramos, Egda M.

    La teoria de evolucion biologica (TEB) por seleccion natural es uno de los conceptos unificadores mas importantes del curriculo de Biologia. En Puerto Rico se han hecho pocas investigaciones que abunden sobre las concepciones y concepciones alternativas (CA) que tienen los estudiantes universitarios/as de Biologia y los maestros/as de Ciencia del nivel secundario sobre esta teoria. La politica publica educativa actual establece mediante documentos normativos como los Estandares de contenido y Expectativas de grado del Programa de Ciencias [Puerto Rico Core Standards] la ensenanza de esta teoria. Sin embargo, no se encontraron preguntas sobre la seleccion natural en los ejercicios de practica provistos por el Departamento de Educacion para las pruebas estandarizadas lo cual puede influir para que no se ensene adecuadamente. Las preguntas de investigacion fueron 1. ¿Cuales son las concepciones y concepciones alternativas de estudiantes universitarios/as y de los futuros maestros y maestras de Ciencia sobre la TEB? 2. ¿Cuales conceptos que seleccionan los estudiantes universitarios/as y los futuros maestros y maestras de Ciencia sobre la TEB coinciden con lo aceptado como valido por la comunidad cientifica? y 3. ¿Como comparan las respuestas de la prueba original. v. Entendiendo el cambio biologico que mide concepciones y CA sobre la TEB por seleccion natural, con las de la traducida al idioma espanol? Se utilizo el metodo cuantitativo con un diseno de investigacion transversal por encuesta. La tecnica principal para recopilar los datos fue una prueba con doce items, que formo parte de un instrumento para el cual se recopilaron diversas fuentes de evidencia acerca de su validez. Las muestras estuvieron formadas por 69 estudiantes de Ciencias Naturales y por 16 estudiantes futuros maestros y maestras del nivel secundario de la UPR-RP. Se utilizaron estadisticas descriptivas, analisis de Ji cuadrado y se calcularon los coeficientes alfa de Cronbach y de Spearman

  19. Estimation of AOT and SDS CMC in a methanol using conductometry, viscometry and pyrene fluorescence spectroscopy methods

    NASA Astrophysics Data System (ADS)

    Mitsionis, Anastasios I.; Vaimakis, Tiverios C.

    2012-09-01

    Critical micelle concentration (CMC) of two anionic surfactants in methanol was estimated using conductometry, viscometry and pyrene fluorescence spectroscopy methods. The surfactants used, were sodium bis(2-ethylhexyl) sulfosuccinate (Aerosol-OT, AOT) and sodium dodecyl sulfate (SDS) dispersed in pure methanol. The CMC determination was evaluated in room temperature. The results have shown nearly similar concentrations.

  20. A novel electrochemical sensor for detecting hyperin with a nanocomposite of ZrO2-SDS-SWCNTs as decoration.

    PubMed

    Li, Shuo; Lei, Sheng; Yu, Qian; Zou, Lina; Ye, Baoxian

    2018-08-01

    A novel high-sensitive electrochemical sensor with glassy carbon electrode (GCE) as support for hyperin determination is successfully designed and constructed, and the well-shaped nano-meter modified material is synthesized via a one-step and facile route. Functionalized with surfactant sodium dodecyl sulfate (SDS), Single-Walled Carbon Nanotubes (SWCNTs) are synchronously grafted with ZrO 2 nanoparticles to develop into the as-prepared nano-composite (ZrO 2 -SDS-SWCNTs). Compared to the previous reports related with hyperin detection, the linear range gets wider and detection limit (LOD) becomes lower with the aid of this novel nano-composite modified glassy carbon electrode (ZrO 2 -SDS-SWCNTs/GCE). The crystalline phases and functionalization of the preparation process has been investigated by X-ray diffraction (XRD) and Fourier transform infrared (FT-IR) instrument analysis, respectively, and the micro-morphology of related modified materials is also visibly characterized by scanning electron microscope (SEM) and transmission electron microscope (TEM). In addition, electrochemical properties of the modified materials are comparably explored by means of impedance spectroscopy (EIS) and cyclic voltammograms (CV). According to the established calibration curve under optimized condition, the peak current (Differential pulse voltammetry (DPV) signal) keeps a linear relationship with hyperin concentration in the ranges of 1.0 × 10 -9 - 3.0 × 10 -7 mol L -1 , meanwhile detection limit reaches as low as 5 × 10 -10 mol L -1 (S/N = 3). As for practical applications, the proposed sensor has also worked well on sensitive hyperin determination in real species Abelmoschus manihot. Copyright © 2018 Elsevier B.V. All rights reserved.

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  2. Poblaciones de los niveles atómicos en condiciones de no equilibrio

    NASA Astrophysics Data System (ADS)

    Cruzado, A.

    El objetivo de este trabajo es encontrar la distribución de los átomos en los diferentes niveles de energía. Con el propósito de encontrar resultados generales y de amplia aplicación, hemos planteado las ecuaciones de equilibrio estadístico como función del número atómico del elemento a considerar y de las condiciones físicas del medio (temperatura y densidad). Así también se ha intentado tener en cuenta todos los niveles atómicos considerando explícitamente aquellos con un número cuántico principal menor que un cierto valor n y calculando una expresión aproximada para estimar la influencia de los demás.

  3. Effects of Microheterogeneity in Hen Egg-White Lysozyme Crystallization

    NASA Technical Reports Server (NTRS)

    Thomas, B. R.; Vekilov, P. G.; Rosenberger, F.

    1998-01-01

    In earlier sodium dodecylsulfate polyacylamide gel electrophoresis (SDS-PAGE) studies it has been found that commonly utilized commercial hen egg-white lysozyme (HEWL) preparations contained 0.2-0.4 mol% covalently bound dimers. Here it is shown, using high-performance capillary electrophoresis (HPCE), that HEWL contains, in addition, two differently charged monomers in comparable amounts. To explore the origin of these microheterogeneous contaminants, purified HEWL (PHEWL) has been oxidized with hydrogen peroxide (0.0026-0.88 M) at various pH levels between 4.5 and 12.0. Optical densitometry of oxidized PHEWL (OHEWL) bands in SDS PAGE gels shows that hydrogen peroxide at 0.88 M in acetate buffer pH 4.5 increased the amount of dimers about sixfold over that in commercial HEWL. OHEWL had, in addition to one of the two monomer forms found in HEWL and PHEWL, three other differently charged monomer forms, each of them representing about 25% of the preparation. SDS-PAGE analysis of OHEWL yielded two closely spaced dimer bands with M(sub r) = 28 000 and 27 500. In addition, larger HEWL oligomers with M, = 1.7 million and 320 000 were detected by gel-filtration fast protein liquid chromatography with multiangle laser light scattering detection. Non-dissociating PAGE in large pore size gels at pH 4.5 confirmed the presence of these large oligomers in HEWL and OHEWL. Increased microheterogeneity resulted in substantial effects on crystal growth and nucleation rate. On addition of 10 microgram(exp -1) mg ml(exp -1) OHEWL to 32 mg ml(exp -1) HEWL crystallizing solutions, both the number and size of forming crystals decreased roughly proportionally to the concentration of the added microheterogeneity. The same effect was observed in HEWL solutions on addition of 0.03-9,3 M Hydrogen peroxide. Repartioning of the dimer during crystallzation aat various temperatures between 277 and 293 K was analyzed by SDS-PAGE. The crystals contained <= 25 % weight by volume of the oligomers in

  4. ATZ11 recognizes not only Z-α1-antitrypsin-polymers and complexed forms of non-Z-α1-antitrypsin but also the von Willebrand factor.

    PubMed

    Goltz, Diane; Hittetiya, Kanishka; Yadegari, Hamideh; Driesen, Julia; Kirfel, Jutta; Neuhaus, Thomas; Steiner, Susanne; Esch, Christiane; Bedorf, Jörg; Hertfelder, Hans-Jörg; Fischer, Hans-Peter

    2014-01-01

    The ATZ11 antibody has been well established for the identification of α1-anti-trypsin (AAT) molecule type PiZ (Z-AAT) in blood samples and liver tissue. In this study, we systematically analyzed the antibody for additional binding sites in human tissue. Ultrastructural ATZ11 binding was investigated immunoelectron microscopically in human umbilical vein endothelial cells (HUVECs) and in platelets of a healthy individual. Human embryonic kidney (HEK293) cells were transiently transfected with Von Willebrand factor (VWF) and analyzed immunocytochemically using confocal microscopy and SDS-PAGE electrophoresis followed by western blotting (WB). Platelets and serum samples of VWF-competent and VWF-deficient patients were investigated using native PAGE and SDS-PAGE electrophoresis followed by WB. The specificity of the ATZ11 reaction was tested immunohistochemically by extensive antibody-mediated blocking of AAT- and VWF-antigens. ATZ11-positive epitopes could be detected in Weibel-Palade bodies (WPBs) of HUVECs and α-granules of platelets. ATZ11 stains pseudo-WBP containing recombinant wild-type VWF (rVWF-WT) in HEK293 cells. In SDS-PAGE electrophoresis followed by WB, anti-VWF and ATZ11 both identified rVWF-WT. However, neither rVWF-WT-multimers, human VWF-multimers, nor serum proteins of VWF-deficient patients were detected using ATZ11 by WB, whereas anti-VWF antibody (anti-VWF) detected rVWF-WT-multimers as well as human VWF-multimers. In human tissue specimens, AAT-antigen blockade using anti-AAT antibody abolished ATZ11 staining of Z-AAT in a heterozygous AAT-deficient patient, whereas VWF-antigen blockade using anti-VWF abolished ATZ11 staining of endothelial cells and megakaryocytes. ATZ11 reacts with cellular bound and denatured rVWF-WT and human VWF as shown using immunocytochemistry and subsequent confocal imaging, immunoelectron microscopy, SDS-PAGE and WB, and immunohistology. These immunoreactions are independent of the binding of Z-AAT-molecules and non

  5. ATZ11 Recognizes Not Only Z-α1-Antitrypsin-Polymers and Complexed Forms of Non-Z-α1-Antitrypsin but Also the von Willebrand Factor

    PubMed Central

    Yadegari, Hamideh; Driesen, Julia; Kirfel, Jutta; Neuhaus, Thomas; Steiner, Susanne; Esch, Christiane; Bedorf, Jörg; Hertfelder, Hans-Jörg; Fischer, Hans-Peter

    2014-01-01

    Aims The ATZ11 antibody has been well established for the identification of α1-anti-trypsin (AAT) molecule type PiZ (Z-AAT) in blood samples and liver tissue. In this study, we systematically analyzed the antibody for additional binding sites in human tissue. Methods and Results Ultrastructural ATZ11 binding was investigated immunoelectron microscopically in human umbilical vein endothelial cells (HUVECs) and in platelets of a healthy individual. Human embryonic kidney (HEK293) cells were transiently transfected with Von Willebrand factor (VWF) and analyzed immunocytochemically using confocal microscopy and SDS-PAGE electrophoresis followed by western blotting (WB). Platelets and serum samples of VWF-competent and VWF-deficient patients were investigated using native PAGE and SDS-PAGE electrophoresis followed by WB. The specificity of the ATZ11 reaction was tested immunohistochemically by extensive antibody-mediated blocking of AAT- and VWF-antigens. ATZ11-positive epitopes could be detected in Weibel-Palade bodies (WPBs) of HUVECs and α-granules of platelets. ATZ11 stains pseudo-WBP containing recombinant wild-type VWF (rVWF-WT) in HEK293 cells. In SDS-PAGE electrophoresis followed by WB, anti-VWF and ATZ11 both identified rVWF-WT. However, neither rVWF-WT-multimers, human VWF-multimers, nor serum proteins of VWF-deficient patients were detected using ATZ11 by WB, whereas anti-VWF antibody (anti-VWF) detected rVWF-WT-multimers as well as human VWF-multimers. In human tissue specimens, AAT-antigen blockade using anti-AAT antibody abolished ATZ11 staining of Z-AAT in a heterozygous AAT-deficient patient, whereas VWF-antigen blockade using anti-VWF abolished ATZ11 staining of endothelial cells and megakaryocytes. Conclusions ATZ11 reacts with cellular bound and denatured rVWF-WT and human VWF as shown using immunocytochemistry and subsequent confocal imaging, immunoelectron microscopy, SDS-PAGE and WB, and immunohistology. These immunoreactions are independent of

  6. Identification of the protective effects of traditional medicinal plants against SDS-induced Drosophila gut damage.

    PubMed

    Zhou, Yang; Liu, Zonglin; Chen, Yuchen; Jin, Li Hua

    2016-10-01

    Traditional medicinal plants are widely used as immunomodulatory medicines that help improve health. A total of 50 different plants used for the treatment of toxicity were screened for their in vivo protective effects. Flies were fed a standard cornmeal-yeast medium (control group) or the standard medium containing medicinal plant extracts (experimental groups). Assessment of the survival rate was performed by feeding flies with toxic compounds. Gut epithelial cells were analyzed for cell proliferation and death by green fluorescent protein antibodies and 7-aminoactinomycin D staining under the microscope. The expression of antimicrobial peptides (AMPs) was evaluated by the quantitative polymerase chain reaction and the results revealed that after feeding the flies with toxic compounds, aqueous extracts from Codonopsis pilosula (Franch.) Nannf ( C. pilosula ), Saussurea lappa (Decne.) C.B.Clarke ( S. lappa ), Imperata cylindrica Beauv.var. major (Nees) C.E. Hubb. ( I. cylindrical var. major ) and Melia toosendan Sied. Et Zucc. ( M.toosendan ) increased the fly survival rate, reduced epithelial cell death and improved gut morphology. In addition, C. pilosula extracts induced the antimicrobial peptide levels (Dpt and Mtk) following treatment with sodium dodecyl sulfate (SDS). However, these extracts were not observed to increase SDS-induced cell proliferation in vivo . These results indicate that there are strong protective effects in extracts of C. pilosula , S. lappa , I. cylindrical var. major and M. toosendan on Drosophila intestinal cells among 50 medicinal plants.

  7. Concentration-dependent antagonistic persuasion of SDS and naphthalene derivatives on the fibrillation of stem bromelain.

    PubMed

    Qadeer, Atiyatul; Ahmad, Ejaz; Zaman, Masihuz; Khan, Mohd Wasif; Khan, Javed Masood; Rabbani, Gulam; Tarique, Khaja Faisal; Sharma, Gaurav; Gourinath, Samudrala; Nadeem, Sajid; Badr, Gamal; Khan, Rizwan Hasan

    2013-12-01

    Sodium dodecyl sulfate, a biological membrane mimetic, can be used to study the conversion of globular proteins into amyloid fibrils in vitro. Using multiple approaches, the effect of SDS was examined on stem bromelain (SB), a widely recognized therapeutic protein. SB is known to exist as a partially folded intermediate at pH 2.0, situation also encountered in the gastrointestinal tract (its site of absorption). In the presence of sub-micellar SDS concentration (500-1000 μM), this intermediate was found to exhibit great propensity to form large-sized β-sheeted aggregates with fibrillar morphology, the hall marks of amyloid structure. We also observed inhibition of fibrillation by two naphthalene-based compounds, ANS and bis-ANS. While bis-ANS significantly inhibited fibril formation at 50 μM, ANS did so at relatively higher concentration (400 μM). Alcohols, but not salts, were found to weaken the inhibitory action of these compounds suggesting the possible involvement of hydrophobic interactions in their binding to protein. Besides, isothermal titration calorimetry and molecular docking studies suggested that inhibition of fibrillation by these naphthalene derivatives is mediated not just through hydrophobic forces, but also by disruption of π-π interactions between the aromatic residues together with the inter-polypeptide chain repulsion among negatively charged ANS/bis-ANS bound SB. Copyright © 2013 Elsevier Inc. All rights reserved.

  8. Description Meta Tags in Public Home and Linked Pages.

    ERIC Educational Resources Information Center

    Craven, Timothy C.

    2001-01-01

    Random samples of 1,872 Web pages registered with Yahoo! And 1,638 pages reachable from Yahoo!-registered pages were analyzed for use of meta tags and specifically those containing descriptions. Results: 727 (38.8%) of the Yahoo!-registered pages and 442 (27%) of the other pages included descriptions in meta tages. Some descriptions greatly…

  9. Tromantadine inhibits HSV-1 induced syncytia formation and viral glycoprotein processing

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Ickes, D.E.

    1989-01-01

    Tromantadine inhibits a late event in Herpes Simplex Virus Type 1 (HSV-1) replication, visualized by the inhibition of both the size and number of syncytia. Tromantadine can be added at any time between 1 and 9 h post infection with complete inhibition of syncytia formation. Glycan synthesis of the viral glycoproteins, important for syncytia formation, is incomplete due to tromantadine treatment. Tromantadine does not inhibit the initiation of glycosylation, since viral glycoproteins, gX{sub t}, synthesized in the presence of tromantadine still incorporate {sup 3}H-glucosamine. Tromantadine does not inhibit the transport of t e viral glycoproteins to the cell surface, sincemore » glycoproteins B, C, and D are expressed, as demonstrated by immunofluorescence. Tromantadine inhibition of HSV-1 glycoprotein processing is demonstrated by an increase in mobility of the radioimmunoprecipitated gX{sub t}, on SDS-PAGE. The gX{sub t} of KOS, a non-syncytial strain of HSV-1, had a similar increase in mobility, suggesting that the block in glycoprotein processing is a general effect of tromantadine treatment. Fucose, which is incorporated into oligosaccharides in the medial Golgi, is incorporated into gX{sub t}, indicating that the tromantadine block in glycoprotein processing occurs after this step. Lectin binding studies and SDS-PAGE analysis of gC processed in the presence of tromantadine, gC{sub t}, indicates that it has terminal galactose residues in both N- and O-linked glycans (binds Peanut and Ricin Agglutinins, respectively). The inhibition of sialylation of N-linked glycans by tromantadine was indicated by the extent of the increase in SDS-PAGE mobility of the G protein from Vesicular Stomatitis Virus. O-glycanase digestion and SDS-PAGE analysis of gC{sub t} indicate that the O-linked disaccharide NAcGal-Galactose is present.« less

  10. Fibrinogenolytic and anticoagulant activities in the tissue covering the stingers of marine stingrays Dasyatis sephen and Aetobatis narinari.

    PubMed

    Kumar, Kalainesan Rajesh; Vennila, Rathinam; Kanchana, Shankar; Arumugam, Muthuvel; Balasubramaniam, Thangavel

    2011-05-01

    Stingray envenomation is one of the major problems in the marine and freshwater ecosystem. Accidents in human cause immediate, local and intense pain, erythema, edema, hemorrhage, tissue necrosis and secondary bacterial infection are also common. To determine the effect of two marine stingray species Dasyatis sephen and Aetobatis narinari venom extract on coagulation, fibrin(ogen)olytic, proteolytic activities. Plasma coagulation, Thrombin catalyzed fibrinocoagulation, Fibrin plate assay, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), substrate SDS-PAGE and thrombin like activity by using chromogenic substrate were used to determine the effect of venom on plasma coagulation, its fibrin(ogen)olytic and proteolytic activity. The results show the presence of fibrin(ogen)olytic, anticoagulant and gelatinolytic activity in both stingray venom extracts. D. sephen venom delays coagulation of citrated plasma more significantly than A. narinari upon using increasing concentration of the venom. The same results were obtained in the fibrinocoagulation assays. SDS-PAGE analysis of fibrinogen and fibrin after incubation with D. sephen and A. narinari venom show fibrin(ogen)olytic activity. Through SDS-PAGE analysis it is confirmed that the delaying in coagulation process by stingray venom is due to its fibrin(ogen)olytic activity and fibrinolytic activity also confirmed through fibrin plate assay. Zymogram analysis shows the presence of array of gelatinolytic and fibrinogenolytic enzymes above 43-276 kDa in the D. sephen and A. narinari venom respectively. Protease inhibitor studies show the serine and metallo proteases are responsible for these activities. From the results, fibrinogenolytic, proteolytic activity of the stingray venom is confirmed, but it has no thrombin like activity and these activities may aid in hemorrhages, tissue necrosis and secondary bacterial infections at the site of envenomation.

  11. Women's Pages or People's Pages: The Production of News for Women in the "Washington Post" in the 1950s.

    ERIC Educational Resources Information Center

    Yang, Mei-ling

    1996-01-01

    Examines the women's pages of the "Washington Post" in the 1950s that were edited by Marie Sauer. States that the newspaper turned down Sauer's request in 1952 to change from traditional women's pages to a unisex "lifestyle" section. Analyzes how women's pages were shaped by factors such as advertising, professional values, and…

  12. Screening of missing proteins in the human liver proteome by improved MRM-approach-based targeted proteomics.

    PubMed

    Chen, Chen; Liu, Xiaohui; Zheng, Weimin; Zhang, Lei; Yao, Jun; Yang, Pengyuan

    2014-04-04

    To completely annotate the human genome, the task of identifying and characterizing proteins that currently lack mass spectrometry (MS) evidence is inevitable and urgent. In this study, as the first effort to screen missing proteins in large scale, we developed an approach based on SDS-PAGE followed by liquid chromatography-multiple reaction monitoring (LC-MRM), for screening of those missing proteins with only a single peptide hit in the previous liver proteome data set. Proteins extracted from normal human liver were separated in SDS-PAGE and digested in split gel slice, and the resulting digests were then subjected to LC-schedule MRM analysis. The MRM assays were developed through synthesized crude peptides for target peptides. In total, the expressions of 57 target proteins were confirmed from 185 MRM assays in normal human liver tissues. Among the proved 57 one-hit wonders, 50 proteins are of the minimally redundant set in the PeptideAtlas database, 7 proteins even have none MS-based information previously in various biological processes. We conclude that our SDS-PAGE-MRM workflow can be a powerful approach to screen missing or poorly characterized proteins in different samples and to provide their quantity if detected. The MRM raw data have been uploaded to ISB/SRM Atlas/PASSEL (PXD000648).

  13. Library links on medical school home pages.

    PubMed

    Thomas, Sheila L

    2011-01-01

    The purpose of this study was to assess the websites of American Association of Medical Colleges (AAMC)-member medical schools for the presence of library links. Sixty-one percent (n = 92) of home pages of the 150 member schools of the AAMC contain library links. For the 58 home pages not offering such links, 50 provided a pathway of two or three clicks to a library link. The absence of library links on 39% of AAMC medical school home pages indicates that the designers of those pages did not consider the library to be a primary destination for their visitors.

  14. Reduction of Portion Size and Eating Rate Is Associated with BMI-SDS Reduction in Overweight and Obese Children and Adolescents: Results on Eating and Nutrition Behaviour from the Observational KgAS Study.

    PubMed

    Torbahn, Gabriel; Gellhaus, Ines; Koch, Benjamin; von Kries, Rüdiger; Obermeier, Viola; Holl, Reinhard W; Fink, Katharina; van Egmond-Fröhlich, Andreas

    2017-01-01

    Treatment of paediatric obesity focuses on changes of nutrition and eating behaviour and physical activity. The evaluation of the patient education programme by KgAS was utilised to analyse the association of changes of portion size, eating rate and dietary habits with BMI-SDS reductions. Patients (n = 297) were examined at the beginning and at the end of treatment and after 1-year follow-up at different out-patient centres. Their parents completed questionnaires including estimation of children's portion size, eating rate and frequency of food intake. Associations of 1- and 2-year changes in BMI-SDS and behaviour were calculated for patients with complete data in BMI-SDS, portion size, eating rate, frequency of green, yellow and red food intake (n = 131) by multiple linear regression models. Significant changes were found in the desired direction for BMI-SDS, portion size, eating rate and the intake of unfavourable red food items both after 1 and 2 years as well as for the consumption of favourable green food items after 1 year. Significant positive associations with BMI-SDS reduction after 1 and 2 years were detected for portion size (Cohen's f2 0.13 and 0.09) and eating rate (Cohen's f2 0.20 and 0.10), respectively. Reduced portion sizes and eating rates are associated with BMI-SDS reduction after 1 and 2 years. These findings suggest to focus on appropriate portion sizes and reduced eating rates in patient education programmes. © 2017 The Author(s) Published by S. Karger GmbH, Freiburg.

  15. World Wide Web Home Page Design: Patterns and Anomalies of Higher Education Library Home Pages.

    ERIC Educational Resources Information Center

    Stover, Mark; Zink, Steven D.

    1996-01-01

    A review of college and university library home pages concluded that many higher education home pages are badly designed, difficult to navigate, and a poor reflection on the institution. The most common shortcoming was the tendency to create too many links or overly large graphics. An appendix lists points to consider when constructing a home…

  16. Identification of the protective effects of traditional medicinal plants against SDS-induced Drosophila gut damage

    PubMed Central

    Zhou, Yang; Liu, Zonglin; Chen, Yuchen; Jin, Li Hua

    2016-01-01

    Traditional medicinal plants are widely used as immunomodulatory medicines that help improve health. A total of 50 different plants used for the treatment of toxicity were screened for their in vivo protective effects. Flies were fed a standard cornmeal-yeast medium (control group) or the standard medium containing medicinal plant extracts (experimental groups). Assessment of the survival rate was performed by feeding flies with toxic compounds. Gut epithelial cells were analyzed for cell proliferation and death by green fluorescent protein antibodies and 7-aminoactinomycin D staining under the microscope. The expression of antimicrobial peptides (AMPs) was evaluated by the quantitative polymerase chain reaction and the results revealed that after feeding the flies with toxic compounds, aqueous extracts from Codonopsis pilosula (Franch.) Nannf (C. pilosula), Saussurea lappa (Decne.) C.B.Clarke (S. lappa), Imperata cylindrica Beauv.var.major (Nees) C.E. Hubb. (I. cylindrical var. major) and Melia toosendan Sied. Et Zucc. (M.toosendan) increased the fly survival rate, reduced epithelial cell death and improved gut morphology. In addition, C. pilosula extracts induced the antimicrobial peptide levels (Dpt and Mtk) following treatment with sodium dodecyl sulfate (SDS). However, these extracts were not observed to increase SDS-induced cell proliferation in vivo. These results indicate that there are strong protective effects in extracts of C. pilosula, S. lappa, I. cylindrical var. major and M. toosendan on Drosophila intestinal cells among 50 medicinal plants. PMID:27698771

  17. Optimizing TLB entries for mixed page size storage in contiguous memory

    DOEpatents

    Chen, Dong; Gara, Alan; Giampapa, Mark E.; Heidelberger, Philip; Kriegel, Jon K.; Ohmacht, Martin; Steinmacher-Burow, Burkhard

    2013-04-30

    A system and method for accessing memory are provided. The system comprises a lookup buffer for storing one or more page table entries, wherein each of the one or more page table entries comprises at least a virtual page number and a physical page number; a logic circuit for receiving a virtual address from said processor, said logic circuit for matching the virtual address to the virtual page number in one of the page table entries to select the physical page number in the same page table entry, said page table entry having one or more bits set to exclude a memory range from a page.

  18. Study of the adsorption of sodium dodecyl sulfate (SDS) at the air/water interface: targeting the sulfate headgroup using vibrational sum frequency spectroscopy.

    PubMed

    Johnson, C Magnus; Tyrode, Eric

    2005-07-07

    The surface sensitive technique vibrational sum frequency spectroscopy (VSFS), has been used to study the adsorption behaviour of SDS to the liquid/vapour interface of aqueous solutions, specifically targeting the sulfate headgroup stretches. In the spectral region extending from 980 to 1850 cm(-1), only the vibrations due to the SO(3) group were detectable. The fitted amplitudes for the symmetric SO(3) stretch observed at 1070 cm(-1) for the polarization combinations ssp and ppp, were seen to follow the adsorption isotherm calculated from surface tension measurements. The orientation of the sulfate headgroup in the concentration range spanning from 1.0 mM to above the critical micellar concentration (c.m.c.) was observed to remain constant within experimental error, with the pseudo-C(3) axis close to the surface normal. Furthermore, the effect of increasing amounts of sodium chloride at SDS concentrations above c.m.c. was also studied, showing an increase of approximately 12% in the fitted amplitude for the symmetric SO(3) stretch when increasing the ionic strength from 0 to 300 mM NaCl. Interestingly, the orientation of the SDS headgroup was also observed to remain constant within this concentration range and identical to the case without NaCl.

  19. Economical and eco-friendly recycling of used dry batteries for synthesis of graphene oxide by sheer exfoliation in presence of SDS

    NASA Astrophysics Data System (ADS)

    Kochrekar, Sachin; Agharkar, Mahesh; Salgaonkar, Manjunath; Gharge, Mrunal; Hidouri, Slah; Azeez, Musibau A.

    2015-06-01

    Graphene is a two-dimensional form of graphite that has attracted great curiosity for its novel physical properties. A key challenge that has emerged is how to create large amounts of graphene at low cost. The purpose of this Paper is to explore a new method to exfoliate graphite extracted from used dry battery in a small scale blender; in presence of SDS surfactant to synthesize graphene oxide, which can be then reduced to graphene. Quantity of SDS required is extremely less (1/10th) of graphite, and it replaces several steps and chemicals such as KMnO4, H2O2, H2SO4 and NaNO3. In this paper, we present the new process and preliminary characterization of synthesized graphene oxide by Raman and UV-Vis absorbance spectroscopy and ATR-IR spectroscopy.

  20. JERHRE's New Web Pages.

    PubMed

    2006-06-01

    JERHRE'S WEBSITE, www.csueastbay.edu/JERHRE/ has two new pages. One of those pages is devoted to curriculum that may be used to educate students, investigators and ethics committee members about issues in the ethics of human subjects research, and to evaluate their learning. It appears at www.csueastbay.edu/JERHRE/cur.html. The other is devoted to emailed letters from readers. Appropriate letters will be posted as soon as they are received by the editor. Letters from readers appear at www.csueastbay.edu/JERHRE/let.html.

  1. Google Analytics: Single Page Traffic Reports

    EPA Pesticide Factsheets

    These are pages that live outside of Google Analytics (GA) but allow you to view GA data for any individual page on either the public EPA web or EPA intranet. You do need to log in to Google Analytics to view them.

  2. Galloylated polyphenols efficiently reduce alpha-tocopherol radicals in a phospholipid model system composed of sodium dodecyl sulfate (SDS) micelles.

    PubMed

    Pazos, Manuel; Torres, Josep Lluís; Andersen, Mogens L; Skibsted, Leif H; Medina, Isabel

    2009-06-10

    The ability of several polyphenolic fractions from grape ( Vitis vinifera ) pomace, pine ( Pinus pinaster ) bark, and witch hazel ( Hammamelis virginiana ) bark to repair alpha-tocopherol (alpha-TOH) through reduction of the alpha-tocopheroxyl radical was investigated in a homogeneous hexane system and a phospholipid-like system based on SDS micelles. These natural polyphenols were compared with pure related phenolics (epicatechin, gallic acid, epigallocatechin gallate, quercetin, and rutin) and ascorbic acid, which is a substance with a well-recognized capacity for regenerating alpha-TOH. alpha-Tocopheroxyl radicals were monitored and quantified by electron spin resonance (ESR) spectroscopy in the absence and presence of phenolics. Polyphenols from grape and pine bark were essentially catechin monomers and proanthocyanidins differing in the content of galloyl residues; those from pine bark had a negligible degree of galloylation. Polyphenolic fractions from witch hazel bark were composed of approximately 80% hydrolyzable tannins rich in galloyl moieties, together with a smaller amount of catechin monomers and proanthocyanidins. In the homogeneous hexane system, polyphenols from grape and pine bark exhibited similar activities, reducing the alpha-tocopheroxyl radicals by over 27-40%, whereas phenols from witch hazel were more highly effective, reducing 80% of alpha-TOH. In contrast, pine bark polyphenols were found to be significantly less active than the grape fractions in SDS micelles, reducing 30 and 70% of alpha-tocopheroxyl radicals, respectively. Polyphenolic fractions from witch hazel were also able to reduce the highest amount of alpha-TOH in SDS-micelles. The reducing capacity on alpha-tocopheroxyl radical of polyphenolic fractions was found to be pH-dependent and more effective at higher pH in the range of pH studied (5.8-7.8). These results stress the potential role of polyphenols, in particular those rich in galloyl groups, to maintain intact endogenous

  3. Dermatology Internet Yellow Page advertising.

    PubMed

    Francis, Shayla; Kozak, Katarzyna Z; Heilig, Lauren; Lundahl, Kristy; Bowland, Terri; Hester, Eric; Best, Arthur; Dellavalle, Robert P

    2006-07-01

    Patients may use Internet Yellow Pages to help select a physician. We sought to describe dermatology Internet Yellow Page advertising. Dermatology advertisements in Colorado, California, New York, and Texas at 3 Yellow Page World Wide Web sites were systematically examined. Most advertisements (76%; 223/292) listed only one provider, 56 listed more than one provider, and 13 listed no practitioner names. Five advertisements listed provider names without any credentialing letters, 265 listed at least one doctor of medicine or osteopathy, and 9 listed only providers with other credentials (6 doctors of podiatric medicine and 3 registered nurses). Most advertisements (61%; 179/292) listed a doctor of medicine or osteopathy claiming board certification, 78% (139/179) in dermatology and 22% (40/179) in other medical specialties. Four (1%; 4/292) claims of board certification could not be verified (one each in dermatology, family practice, dermatologic/cosmetologic surgery, and laser surgery). Board certification could be verified for most doctors of medicine and osteopathy not advertising claims of board certification (68%; 41/60; 32 dermatology, 9 other specialties). A total of 50 advertisements (17%) contained unverifiable or no board certification information, and 47 (16%) listed a physician with verifiable board certification in a field other than dermatology. All Internet Yellow Page World Wide Web sites and all US states were not examined. Nonphysicians, physicians board certified in medical specialties other than dermatology, and individuals without verifiable board certification in any medical specialty are advertising in dermatology Internet Yellow Pages. Many board-certified dermatologists are not advertising this certification.

  4. Extracellular metalloproteinases in Phytomonas serpens.

    PubMed

    Vermelho, Alane B; Almeida, Flávia V S; Bronzato, Leandro S; Branquinha, Marta H

    2003-03-01

    The detection of extracellular proteinases in Phytomonas serpens, a trypanosomatid isolated from tomato fruits, is demonstrated in this paper. Maximal production occurred at the end of the logarithmic phase of growth. These enzymes exhibited selective substrate utilization in SDS-PAGE, being more active with gelatin; hemoglobin and bovine serum albumin were not degraded. Three proteinases were detected in SDS-PAGE-gelatin, with apparent molecular masses between 94 and 70 kDa. The proteolytic activity was completely blocked by 1,10-phenanthroline and strongly inhibited by EDTA, whereas a partial inhibition was observed with trans-epoxysuccinyl-L-leucylamido-(4-guanidino) butane (E-64) and soybean trypsin inhibitor; phenylmethylsulfonyl fluoride weakly inhibited the enzymes. This inhibition profile indicated that these extracellular proteinases belong to the metalloproteinase class.

  5. Assays for Determination of Protein Concentration.

    PubMed

    Olson, Bradley J S C

    2016-06-01

    Biochemical analysis of proteins relies on accurate quantification of protein concentration. Detailed in this appendix are some commonly used methods for protein analysis, e.g., Lowry, Bradford, bicinchoninic acid (BCA), UV spectroscopic, and 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA) assays. The primary focus of this report is assay selection, emphasizing sample and buffer compatibility. The fundamentals of generating protein assay standard curves and of data processing are considered, as are high-throughput adaptations of the more commonly used protein assays. Also included is a rapid, inexpensive, and reliable BCA assay of total protein in SDS-PAGE sample buffer that is used for equal loading of SDS-PAGE gels. © 2016 by John Wiley & Sons, Inc. Copyright © 2016 John Wiley & Sons, Inc.

  6. The (Untold) Drama of the Turning Page: The Role of Page Breaks in Understanding Picture Books

    ERIC Educational Resources Information Center

    Jacobs, Katrina Emily Bartow

    2016-01-01

    While scholars have recognized the importance of page breaks in both the construction and comprehension of narrative within picture books, there has previously been limited research that focused directly on how children discuss and make sense of these spaces in the text. Yet, because of their nature as dramatic gaps in the narrative, page breaks…

  7. Coupling Sodium Dodecyl Sulfate–Capillary Polyacrylamide Gel Electrophoresis with MALDI-TOF-MS via a PTFE Membrane

    PubMed Central

    Lu, Joann J.; Zhu, Zaifang; Wang, Wei; Liu, Shaorong

    2011-01-01

    Sodium dodecyl sulfate (SDS)–polyacrylamide gel electrophoresis (PAGE) is a fundamental analytical technique for proteomic research, and SDS–capillary gel electrophoresis (CGE) is its miniaturized version. Compared to conventional slab-gel electrophoresis, SDS-CGE has many advantages such as increased separation efficiency, reduced separation time and automated operation. SDS-CGE is not widely accepted in proteomic research primarily due to the difficulties in identifying the well-resolved proteins. MALDI–TOF–MS is an outstanding platform for protein identifications. Coupling the two would solve the problem but is extremely challenging because the MS detector has no access to the SDS-CGE resolved proteins and the SDS interferes with MS detection. In this work we introduce an approach to address these issues. We discover that poly(tetrafluoroethylene) (PTFE) membranes are excellent materials for collecting SDS-CGE separated proteins. We demonstrate that we can wash off the SDS bound to the collected proteins and identify these proteins on-membrane with MALDI-TOF-MS. We also show that we can immunoblot and Coomassie-stain the proteins collected on these membranes. PMID:21309548

  8. Characteristics of emergency pages using a computer-based anesthesiology paging system in children and adults undergoing procedures at a tertiary care medical center.

    PubMed

    Weingarten, Toby N; Abenstein, John P; Dutton, Claire H; Kohn, Melinda A; Lee, Elizabeth A; Mullenbach, Tami E; Narr, Bradly J; Schroeder, Darrell R; Sprung, Juraj

    2013-04-01

    In our large academic supervisory practice, attending anesthesiologists concomitantly care for multiple patients. To manage communications within the procedural environment, we use a proprietary electronic computer-based anesthesiology visual paging system. This system can send an emergency page that instantly alerts the attending anesthesiologist and other available personnel that immediate help is needed. We analyzed the characteristics of intraoperative emergency pages in children and adults. We identified all emergency page activations between January 1, 2005 and July 31, 2010 in our main operating rooms. Electronic medical records were reviewed for rates and characteristics of pages such as primary etiology, performed interventions, and outcomes. During the study period, 258,135 anesthetics were performed (n = 32,103 children, younger than 18 years) and 370 emergency pages (n = 309 adults, n = 61 children) were recorded (1.4 per 1000 cases; 95% confidence interval, 1.3-1.6). Infants had the highest rates (9.4 per 1000; 95% confidence interval, 5.7-14.4) of emergency page activations (P < 0.001 compared with each other age group). In adults, the most frequent causes were hemodynamic (55%), and in children respiratory and airway (60.7%) events. Emergency pages were rare in patients older than 2 years. Infants were more likely than children 1 to 2 years of age to have emergency page activation, despite both groups being cared for by pediatric fellowship trained anesthesiologists.

  9. Protein changes in leaf-sheath pulvini of barley (hordeum) induced by gravistimulation

    NASA Technical Reports Server (NTRS)

    Kaufman, P. B.; Song, I.

    1984-01-01

    Sodium dodecyl sulfate polyacrylamade gel electrophoresis (SDS-PAGE) pattern of salt soluble proteins elicite by gravistimulation were shown in the top and the bottom halves of the gravistimulated pulvini as follows: at least five proteins were increased in the tissues derived from the bottom halves of the pulvini in the approximate molecular weight range of 91, 57, 50, 22, 17 kilodatons. SDS densitometric scans indicated that the two of them are probably cellulase and invertase.

  10. Page Recognition: Quantum Leap In Recognition Technology

    NASA Astrophysics Data System (ADS)

    Miller, Larry

    1989-07-01

    No milestone has proven as elusive as the always-approaching "year of the LAN," but the "year of the scanner" might claim the silver medal. Desktop scanners have been around almost as long as personal computers. And everyone thinks they are used for obvious desktop-publishing and business tasks like scanning business documents, magazine articles and other pages, and translating those words into files your computer understands. But, until now, the reality fell far short of the promise. Because it's true that scanners deliver an accurate image of the page to your computer, but the software to recognize this text has been woefully disappointing. Old optical-character recognition (OCR) software recognized such a limited range of pages as to be virtually useless to real users. (For example, one OCR vendor specified 12-point Courier font from an IBM Selectric typewriter: the same font in 10-point, or from a Diablo printer, was unrecognizable!) Computer dealers have told me the chasm between OCR expectations and reality is so broad and deep that nine out of ten prospects leave their stores in disgust when they learn the limitations. And this is a very important, very unfortunate gap. Because the promise of recognition -- what people want it to do -- carries with it tremendous improvements in our productivity and ability to get tons of written documents into our computers where we can do real work with it. The good news is that a revolutionary new development effort has led to the new technology of "page recognition," which actually does deliver the promise we've always wanted from OCR. I'm sure every reader appreciates the breakthrough represented by the laser printer and page-makeup software, a combination so powerful it created new reasons for buying a computer. A similar breakthrough is happening right now in page recognition: the Macintosh (and, I must admit, other personal computers) equipped with a moderately priced scanner and OmniPage software (from Caere

  11. Page turning solutions for musicians: a survey.

    PubMed

    Wolberg, George; Schipper, Irene

    2012-01-01

    Musicians have long been hampered by the challenge in turning sheet music while their hands are occupied playing an instrument. The sight of a human page turner assisting a pianist during a performance, for instance, is not uncommon. This need for a page turning solution is no less acute during practice sessions, which account for the vast majority of playing time. Despite widespread appreciation of the problem, there have been virtually no robust and affordable products to assist the musician. Recent progress in assistive technology and electronic reading devices offers promising solutions to this long-standing problem. The objective of this paper is to survey the technology landscape and assess the benefits and drawbacks of page turning solutions for musicians. A full range of mechanical and digital page turning products are reviewed.

  12. Neutron Scattering Home Page (Low-Graphics)

    Science.gov Websites

    will be added. We encourage everyone interested in neutron scattering to take full advantage of this Home Page | Facilities | Reference | Software | Conferences | Announcements | Mailing Lists Neutron Scattering Banner Neutron Scattering Home Page A new portal for neutron scattering has just been established

  13. Home Page, Sweet Home Page: Creating a Web Presence.

    ERIC Educational Resources Information Center

    Falcigno, Kathleen; Green, Tim

    1995-01-01

    Focuses primarily on design issues and practical concerns involved in creating World Wide Web documents for use within an organization. Concerns for those developing Web home pages are: learning HyperText Markup Language (HTML); defining customer group; allocating staff resources for maintenance of documents; providing feedback mechanism for…

  14. Educational use of World Wide Web pages on CD-ROM.

    PubMed

    Engel, Thomas P; Smith, Michael

    2002-01-01

    The World Wide Web is increasingly important for medical education. Internet served pages may also be used on a local hard disk or CD-ROM without a network or server. This allows authors to reuse existing content and provide access to users without a network connection. CD-ROM offers several advantages over network delivery of Web pages for several applications. However, creating Web pages for CD-ROM requires careful planning. Issues include file names, relative links, directory names, default pages, server created content, image maps, other file types and embedded programming. With care, it is possible to create server based pages that can be copied directly to CD-ROM. In addition, Web pages on CD-ROM may reference Internet served pages to provide the best features of both methods.

  15. High-pressure-induced interactions between milk fat globule membrane proteins and skim milk proteins in whole milk.

    PubMed

    Ye, A; Anema, S G; Singh, H

    2004-12-01

    The association of beta-lactoglobulin (beta-LG) and alpha-lactalbumin (alpha-LA) with milk fat globule membrane (MFGM), when whole milk was treated by high pressure in the range 100 to 800 MPa, was investigated using sodium dodecyl sulfate (SDS)-PAGE under reducing and nonreducing conditions. In SDS-PAGE under reducing conditions, beta-LG was observed in the MFGM material isolated from milk treated at 100 to 800 MPa for 30 min, and small amounts of alpha-LA and kappa-casein were also observed at pressures >600 MPa for 30 min. However, these proteins were not observed in SDS-PAGE under nonreducing conditions. These results indicate that beta-LG and alpha-LA associated with MFGM proteins via disulfide bonds during the high-pressure treatment of whole milk. The amount of beta-LG associated with the MFGM increased with an increase in pressure up to 800 MPa and with increasing time of pressure treatment. The maximum value for beta-LG association with the MFGM was approximately 0.75 mg/g of fat. Of the major original MFGM proteins, no change in butyrophilin was observed during the high-pressure treatment of whole milk, whereas xanthine oxidase was reduced to some extent beyond 400 MPa. In contrast to the behavior during heat treatment, PAS 6 and PAS 7 were stable during high-pressure treatment, and they remained associated with the MFGM.

  16. Identification and partial purification of pollen allergens from Artemisia princeps.

    PubMed

    Park, H S; Hong, C S; Choi, H J; Hahm, K S

    1989-12-01

    The pollen of Artemisia has been considered as the main late summer-autumn allergen source in this country. To identify its allergenic components, Artemisia princeps pollen extracts were separated by 10% sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE), and transferred to nitrocellulose membrane, where IgE binding components were detected by the reaction with sera of twenty Artemisia-allergic patients and 125I-anti-human IgE, sixteen components in the molecular range of 10,000 and 85,000 daltons were detected. Twelve bands bound to IgE from 50% of the sera tested, and two bands (37,000, 23,000 daltons) showed the highest (85%) frequency of IgE-binding in twenty sera tested. When the gel of SDS-PAGE with Artemisia pollen extracts was sliced into 11 allergenic groups (AG) and the protein of each AG was obtained by the gel elution method, the wormwool-RAST inhibition test showed that the AG 10 demonstrated to be the most potent, and the AG 7 was the next. Six AGs showed significant responses (more than 100% of wheal size to histamine, 1 mg/ml) on the skin prick test in more than 50% of the patients tested. It is suggested that electrophoretic transfer analysis with SDS-PAGE may be a valuable method for Artemisia allergen identification, and the possibility of partial purification of allergens by employing gel elution is discussed.

  17. Protein profiling of water and alkali soluble cottonseed protein isolates.

    PubMed

    He, Zhongqi; Zhang, Dunhua; Cao, Heping

    2018-06-18

    Currently, there is only limited knowledge on the protein types and structures of the cottonseed proteins. In this work, water-soluble cottonseed proteins (CSPw) and alkali-soluble cottonseed proteins (CSPa) were sequentially extracted from defatted cottonseed meal. Proteins of the two fractions were separated by 4-20% gradient polyacrylamide gel electrophoresis (SDS-PAGE); There were 7 and 12 polypeptide bands on SDS-PAGE of CSPa and CSPw, respectively. These individual bands were then excised from the gel and subjected to mass spectrometric analysis. There were total 70 polypeptides identified from the proteins of the two cottonseed preparations, with molecular weights ranging from 10 to 381 kDa. While many proteins or their fragments were found in multiple bands, 18 proteins appeared only in one SDS-PAGE band (6 in CSPa, 12 in CSPw). Putative functions of these proteins include storage, transcription/translation, synthesis, energy metabolism, antimicrobial activity, and embryogenesis. Among the most abundant are legumin A (58 kDa), legumin B (59 kDa), vicilin C72 (70 kDa), vicilin GC72-A (71 kDa), and vicilin-like antimicrobial peptides (62 kDa). This work enriched the fundamental knowledge on cottonseed protein composition, and would help in better understanding of the functional and physicochemical properties of cottonseed protein and for enhancing its biotechnological utilization.

  18. Identification, purification, and expression patterns of chitinase from psychrotolerant Pedobacter sp. PR-M6 and antifungal activity in vitro.

    PubMed

    Song, Yong-Su; Seo, Dong-Jun; Jung, Woo-Jin

    2017-06-01

    In this study, a novel psychrotolerant chitinolytic bacterium Pedobacter sp. PR-M6 that displayed strong chitinolytic activity on 0.5% colloidal chitin was isolated from the soil of a decayed mushroom. Chitinase activity of PR-M6 at 25 °C (C25) after 6 days of incubation with colloidal chitin increased rapidly to a maximum level (31.3 U/mg proteins). Three chitinase isozymes (chiII, chiIII, and chiIV) from the crude enzyme at 25 °C (C25) incubation were expressed on SDS-PAGE gels at 25 °C. After purification by chitin-affinity chromatography, six chitinase isozymes (chiI, chiII, chiIII, chiIV, chiV, and chiVI) from C25-fractions were expressed on SDS-PAGE gels at 25 °C. Major bands of chitinase isozymes (chiI, chiII, and chiIII) from C4-fractions were strongly expressed on SDS-PAGE gels at 25 °C. Pedobacter sp. PR-M6 showed high inhibition rate of 60.9% and 57.5% against Rhizoctonia solani and Botrytis cinerea, respectively. These results indicated that psychrotolerant Pedobacter sp. PR-M6 could be applied widely as a microorganism agent for the biocontrol of agricultural phytopathogens at low temperatures. Copyright © 2017 Elsevier Ltd. All rights reserved.

  19. Cross-Linking Proteins To Show Complex Formation: A Laboratory That Visually Demonstrates Calmodulin Binding to Calmodulin Kinase II.

    ERIC Educational Resources Information Center

    Porta, Angela R.

    2003-01-01

    Presents a laboratory experiment demonstrating the binding of calcium/calmodulin to calmodulin kinase II, which is important in the metabolic and physiological activities of the cell. Uses SDS polyacrylamide gel electrophoresis (PAGE). (YDS)

  20. Teaching the Structure of Immunoglobulins by Molecular Visualization and SDS-PAGE Analysis

    ERIC Educational Resources Information Center

    Rižner, Tea Lanišnik

    2014-01-01

    This laboratory class combines molecular visualization and laboratory experimentation to teach the structure of the immunoglobulins (Ig). In the first part of the class, the three-dimensional structures of the human IgG and IgM molecules available through the RCSB PDB database are visualized using freely available software. In the second part, IgG…

  1. Assessment of the floral origin of honey by SDS-page immunoblot techniques.

    PubMed

    Baroni, María V; Chiabrando, Gustavo A; Costa, Cristina; Wunderlin, Daniel A

    2002-03-13

    We report on the development of a novel alternative method for the assessment of floral origin in honey samples based on the study of honey proteins using immunoblot assays. The main goal of our work was to evaluate the use of honey proteins as chemical markers of the floral origin of honey. Considering that honeybee proteins should be common to all types of honey, we decided to verify the usefulness of pollen proteins as floral origin markers in honey. We used polyclonal anti-pollen antibodies raised in rabbits by repeated immunization of Sunflower (Elianthus annuus) and Eucalyptus (Eucalyptus sp.) pollen extracts. The IgG fraction was purified by immunoaffinity. These antibodies were verified with nitrocellulose blotted pollen and unifloral honey protein extracts. The antibodies anti-Sunflower pollen, bound to the 36 and 33 kDa proteins of Sunflower unifloral honey and to honey containing Sunflower pollen; and the antibodies anti-Eucalyptus sp. pollen bound to the 38 kDa proteins of Eucalyptus sp. unifloral honey in immunoblot assays. Satisfactory results were obtained in differentiating between the types of pollen analyzed and between Sunflower honey and Eucalyptus honey with less cross reactivity with other types of honey from different origin and also with good sensitivity in the detection. This immunoblot method opens an interesting field for the development of new antibodies from different plants, which could serve as an alternative or complementary method to the usual melissopalynological analysis to assess honey floral origin.

  2. Automatic page layout using genetic algorithms for electronic albuming

    NASA Astrophysics Data System (ADS)

    Geigel, Joe; Loui, Alexander C. P.

    2000-12-01

    In this paper, we describe a flexible system for automatic page layout that makes use of genetic algorithms for albuming applications. The system is divided into two modules, a page creator module which is responsible for distributing images amongst various album pages, and an image placement module which positions images on individual pages. Final page layouts are specified in a textual form using XML for printing or viewing over the Internet. The system makes use of genetic algorithms, a class of search and optimization algorithms that are based on the concepts of biological evolution, for generating solutions with fitness based on graphic design preferences supplied by the user. The genetic page layout algorithm has been incorporated into a web-based prototype system for interactive page layout over the Internet. The prototype system is built using client-server architecture and is implemented in java. The system described in this paper has demonstrated the feasibility of using genetic algorithms for automated page layout in albuming and web-based imaging applications. We believe that the system adequately proves the validity of the concept, providing creative layouts in a reasonable number of iterations. By optimizing the layout parameters of the fitness function, we hope to further improve the quality of the final layout in terms of user preference and computation speed.

  3. Enzymatic hydrolysis of sodium dodecyl sulphate (SDS)-pretreated newspaper for cellulosic ethanol production by Saccharomyces cerevisiae and Pichia stipitis.

    PubMed

    Xin, Fengxue; Geng, Anli; Chen, Ming Li; Gum, Ming Jun Marcus

    2010-10-01

    Fermentation of enzymatic hydrolysate of waste newspaper was investigated for cellulosic ethanol production in this study. Various nonionic and ionic surfactants were applied for waste newspaper pretreatment to increase the enzymatic digestibility. The surfactant-pretreated newspaper was enzymatically digested in 0.05 M sodium citrate buffer (pH 4.8) with varying solid content, filter paper unit loading (FPU/g newspaper), and ratio of filter paper unit/beta-glucosidase unit (FPU/CBU). Newspaper pretreated with the anionic surfactant sodium dodecyl sulphate (SDS) demonstrated the highest sugar yield. The addition of Tween-80 in the enzymatic hydrolysis process enhanced the enzymatic digestibility of newspaper pretreated with all of the surfactants. Enzymatic hydrolysis of SDS-pretreated newspaper with 15% solid content, 15 FPU/g newspaper, and FPU/CBU of 1:4 resulted in a newspaper hydrolysate conditioning 29.07 g/L glucose and 4.08 g/L xylose after 72 h of incubation at 50 degrees C. The fermentation of the enzymatic hydrolysate with Saccharomyces cerevisiae, Pichia stipitis, and their co-culture produced 14.29, 13.45, and 14.03 g/L of ethanol, respectively. Their corresponding ethanol yields were 0.43, 0.41, and 0.42 g/g.

  4. [Purification of arsenic-binding proteins in hamster plasma after oral administration of arsenite].

    PubMed

    Wang, Wenwen; Zhang, Min; Li, Chunhui; Qin, Yingjie; Hua, Naranmandura

    2013-01-01

    To purify the arsenic-binding proteins (As-BP) in hamster plasma after a single oral administration of arsenite (iAs(III)). Arsenite was given to hamsters in a single dose. Three types of HPLC columns, size exclusion, gel filtration and anion exchange columns, combined with an inductively coupled argon plasma mass spectrometer (ICP MS) were used to purify the As-BP in hamster plasma. SDS-PAGE was used to confirm the arsenic-binding proteins at each purification step. The three-step purification process successfully separated As-BP from other proteins (ie, arsenic unbound proteins) in hamster plasma. The molecular mass of purified As-BP in plasma was approximately 40-50 kD on SDS-PAGE. The three-step purification method is a simple and fast approach to purify the As-BP in plasma samples.

  5. NCEP MMAB Sea Ice Home Page

    Science.gov Websites

    NCEP MMAB Sea Ice Home Page The Polar and Great Lakes Ice group works on sea ice analysis from satellite, sea ice modeling, and ice-atmosphere-ocean coupling. Our work supports the Alaska Region of the @noaa.gov Last Modified 2 July 2012 Pages of Interest Analysis Daily Sea Ice Analyses Animations of the

  6. Classifying Web Pages by Using Knowledge Bases for Entity Retrieval

    NASA Astrophysics Data System (ADS)

    Kiritani, Yusuke; Ma, Qiang; Yoshikawa, Masatoshi

    In this paper, we propose a novel method to classify Web pages by using knowledge bases for entity search, which is a kind of typical Web search for information related to a person, location or organization. First, we map a Web page to entities according to the similarities between the page and the entities. Various methods for computing such similarity are applied. For example, we can compute the similarity between a given page and a Wikipedia article describing a certain entity. The frequency of an entity appearing in the page is another factor used in computing the similarity. Second, we construct a directed acyclic graph, named PEC graph, based on the relations among Web pages, entities, and categories, by referring to YAGO, a knowledge base built on Wikipedia and WordNet. Finally, by analyzing the PEC graph, we classify Web pages into categories. The results of some preliminary experiments validate the methods proposed in this paper.

  7. Digital Ethnography: Library Web Page Redesign among Digital Natives

    ERIC Educational Resources Information Center

    Klare, Diane; Hobbs, Kendall

    2011-01-01

    Presented with an opportunity to improve Wesleyan University's dated library home page, a team of librarians employed ethnographic techniques to explore how its users interacted with Wesleyan's current library home page and web pages in general. Based on the data that emerged, a group of library staff and members of the campus' information…

  8. Web page sorting algorithm based on query keyword distance relation

    NASA Astrophysics Data System (ADS)

    Yang, Han; Cui, Hong Gang; Tang, Hao

    2017-08-01

    In order to optimize the problem of page sorting, according to the search keywords in the web page in the relationship between the characteristics of the proposed query keywords clustering ideas. And it is converted into the degree of aggregation of the search keywords in the web page. Based on the PageRank algorithm, the clustering degree factor of the query keyword is added to make it possible to participate in the quantitative calculation. This paper proposes an improved algorithm for PageRank based on the distance relation between search keywords. The experimental results show the feasibility and effectiveness of the method.

  9. CrazyEgg Reports for Single Page Analysis

    EPA Pesticide Factsheets

    CrazyEgg provides an in depth look at visitor behavior on one page. While you can use GA to do trend analysis of your web area, CrazyEgg helps diagnose the design of a single Web page by visually displaying all visitor clicks during a specified time.

  10. World Wide Web Pages--Tools for Teaching and Learning.

    ERIC Educational Resources Information Center

    Beasley, Sarah; Kent, Jean

    Created to help educators incorporate World Wide Web pages into teaching and learning, this collection of Web pages presents resources, materials, and techniques for using the Web. The first page focuses on tools for teaching and learning via the Web, providing pointers to sites containing the following: (1) course materials for both distance and…

  11. Purification and partial characterization of the major outer membrane protein of Chlamydia trachomatis

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Caldwell, H.D.; Kromhout, J.; Schachter, J.

    1981-03-01

    Elementary bodies (EB) of Chlamydia trachomatis serotypes C, E, and L2 were extrinsically radioiodinated, and whole-cell lysates of these serotypes were compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Autoradiography of the polypeptide profiles identified a major surface protein with an apparent subunit molecular weight of 39,500 that was common to each C. trachomatis serotype. The abilities of nonionic (Triton X-100), dipolar ionic (Zwittergent TM-314), mild (sodium deoxycholate and sodium N-lauroyl sarcosine), and strongly anionic (SDS) detergents to extract this protein from intact EB of the L2 serotype were investigated by SDS-PAGE analysis of the soluble and insoluble fractions obtainedmore » after each detergent treatment. Only SDS readily extracted this protein from intact EB. Sarkosyl treatment selectively solubilized the majority of other EB proteins, leaving the 39,500-dalton protein associated with the Sarkosyl-insoluble fraction. Ultrastructural studies of the Sarkosyl-insoluble EB pellet showed it to consist of empty EB particles possessing an apparently intact outer membrane. No structural evidence for a peptidoglycan-like cell wall was found. Morphologically these chlamydial outer membrane complexes (COMC) resembled intact chlamydial EB outer membranes. The 39,500-dalton outer membrane protein was quantitatively extracted from COMC by treating them with 2% SDS at 60 degrees C. This protein accounted for 61% of the total COMC-associated protein, and its extraction resulted in a concomitant loss of the COMC membrane structure and morphology. The 39,500-dalton major outer membrane protein is a serogroup antigen of C. trachomatis organisms.« less

  12. Using shadow page cache to improve isolated drivers performance.

    PubMed

    Zheng, Hao; Dong, Xiaoshe; Wang, Endong; Chen, Baoke; Zhu, Zhengdong; Liu, Chengzhe

    2015-01-01

    With the advantage of the reusability property of the virtualization technology, users can reuse various types and versions of existing operating systems and drivers in a virtual machine, so as to customize their application environment. In order to prevent users' virtualization environments being impacted by driver faults in virtual machine, Chariot examines the correctness of driver's write operations by the method of combining a driver's write operation capture and a driver's private access control table. However, this method needs to keep the write permission of shadow page table as read-only, so as to capture isolated driver's write operations through page faults, which adversely affect the performance of the driver. Based on delaying setting frequently used shadow pages' write permissions to read-only, this paper proposes an algorithm using shadow page cache to improve the performance of isolated drivers and carefully study the relationship between the performance of drivers and the size of shadow page cache. Experimental results show that, through the shadow page cache, the performance of isolated drivers can be greatly improved without impacting Chariot's reliability too much.

  13. Using Shadow Page Cache to Improve Isolated Drivers Performance

    PubMed Central

    Dong, Xiaoshe; Wang, Endong; Chen, Baoke; Zhu, Zhengdong; Liu, Chengzhe

    2015-01-01

    With the advantage of the reusability property of the virtualization technology, users can reuse various types and versions of existing operating systems and drivers in a virtual machine, so as to customize their application environment. In order to prevent users' virtualization environments being impacted by driver faults in virtual machine, Chariot examines the correctness of driver's write operations by the method of combining a driver's write operation capture and a driver's private access control table. However, this method needs to keep the write permission of shadow page table as read-only, so as to capture isolated driver's write operations through page faults, which adversely affect the performance of the driver. Based on delaying setting frequently used shadow pages' write permissions to read-only, this paper proposes an algorithm using shadow page cache to improve the performance of isolated drivers and carefully study the relationship between the performance of drivers and the size of shadow page cache. Experimental results show that, through the shadow page cache, the performance of isolated drivers can be greatly improved without impacting Chariot's reliability too much. PMID:25815373

  14. Classroom Web Pages: A "How-To" Guide for Educators.

    ERIC Educational Resources Information Center

    Fehling, Eric E.

    This manual provides teachers, with very little or no technology experience, with a step-by-step guide for developing the necessary skills for creating a class Web Page. The first part of the manual is devoted to the thought processes preceding the actual creation of the Web Page. These include looking at other Web Pages, deciding what should be…

  15. Purification and characterization of polyphenol oxidase from banana (Musa sapientum L.) pulp.

    PubMed

    Yang, C P; Fujita, S; Ashrafuzzaman, M; Nakamura, N; Hayashi, N

    2000-07-01

    Polyphenol oxidase (EC 1.10.3.1, PPO) in the pulp of banana (Musa sapientum L.) was purified to 636-fold with a recovery of 3.0%, using dopamine as substrate. The purified enzyme exhibited a clear single band on polyacrylamide gel electrophoresis (PAGE) and sodium dodecyl sulfate (SDS)-PAGE. The molecular weight of the enzyme was estimated to be about 41000 and 42000 by gel filtration and SDS-PAGE, respectively. The enzyme quickly oxidized dopamine, and its K(m) value for dopamine was 2.8 mM. The optimum pH was at 6.5, and the enzyme activity was stable in the range of pH 5-11 at 5 degrees C for 48 h. The enzyme had an optimum temperature of 30 degrees C and was stable even after a heat treatment at 70 degrees C for 30 min. The enzyme activity was completely inhibited by L-ascorbic acid, cysteine, sodium diethyldithiocarbamate, and potassium cyanide. Under a low buffer capacity, the enzyme was also strongly inhibited by citric acid and acetic acid at 10 mM.

  16. Recognition of pornographic web pages by classifying texts and images.

    PubMed

    Hu, Weiming; Wu, Ou; Chen, Zhouyao; Fu, Zhouyu; Maybank, Steve

    2007-06-01

    With the rapid development of the World Wide Web, people benefit more and more from the sharing of information. However, Web pages with obscene, harmful, or illegal content can be easily accessed. It is important to recognize such unsuitable, offensive, or pornographic Web pages. In this paper, a novel framework for recognizing pornographic Web pages is described. A C4.5 decision tree is used to divide Web pages, according to content representations, into continuous text pages, discrete text pages, and image pages. These three categories of Web pages are handled, respectively, by a continuous text classifier, a discrete text classifier, and an algorithm that fuses the results from the image classifier and the discrete text classifier. In the continuous text classifier, statistical and semantic features are used to recognize pornographic texts. In the discrete text classifier, the naive Bayes rule is used to calculate the probability that a discrete text is pornographic. In the image classifier, the object's contour-based features are extracted to recognize pornographic images. In the text and image fusion algorithm, the Bayes theory is used to combine the recognition results from images and texts. Experimental results demonstrate that the continuous text classifier outperforms the traditional keyword-statistics-based classifier, the contour-based image classifier outperforms the traditional skin-region-based image classifier, the results obtained by our fusion algorithm outperform those by either of the individual classifiers, and our framework can be adapted to different categories of Web pages.

  17. Enzyme and Microbial Development | Bioenergy | NREL

    Science.gov Websites

    samples of powdered biomass into a reactor plate, part of the high-throughput biomass recalcitrance the molecular weight of cellA on blue and white containers of SDS PAGE gel in a laboratory

  18. Automated Title Page Cataloging: A Feasibility Study.

    ERIC Educational Resources Information Center

    Weibel, Stuart; And Others

    1989-01-01

    Describes the design of a prototype rule-based system for the automation of descriptive cataloging from title pages. The discussion covers the results of tests of the prototype, major impediments to automatic cataloging from title pages, and prospects for further progress. The rules implemented in the prototype are appended. (16 references)…

  19. Exploring the use of a Facebook page in anatomy education.

    PubMed

    Jaffar, Akram Abood

    2014-01-01

    Facebook is the most popular social media site visited by university students on a daily basis. Consequently, Facebook is the logical place to start with for integrating social media technologies into education. This study explores how a faculty-administered Facebook Page can be used to supplement anatomy education beyond the traditional classroom. Observations were made on students' perceptions and effectiveness of using the Page, potential benefits and challenges of such use, and which Insights metrics best reflect user's engagement. The Human Anatomy Education Page was launched on Facebook and incorporated into anatomy resources for 157 medical students during two academic years. Students' use of Facebook and their perceptions of the Page were surveyed. Facebook's "Insights" tool was also used to evaluate Page performance during a period of 600 days. The majority of in-class students had a Facebook account which they adopted in education. Most students perceived Human Anatomy Education Page as effective in contributing to learning and favored "self-assessment" posts. The majority of students agreed that Facebook could be a suitable learning environment. The "Insights" tool revealed globally distributed fans with considerable Page interactions. The use of a faculty-administered Facebook Page provided a venue to enhance classroom teaching without intruding into students' social life. A wider educational use of Facebook should be adopted not only because students are embracing its use, but for its inherent potentials in boosting learning. The "Insights" metrics analyzed in this study might be helpful when establishing and evaluating the performance of education-oriented Facebook Pages. © 2013 American Association of Anatomists.

  20. Characterization of seeds of selected wild species of rice (Oryza) stored under high temperature and humidity conditions.

    PubMed

    Das, Smruti; Nayak, Monalisa; Patra, B C; Ramakrishnan, B; Krishnan, P

    2010-06-01

    Wild progenitors of rice (Oryza) are an invaluable resource for restoring genetic diversity and incorporating useful traits back into cultivars. Studies were conducted to characterize the biochemical changes, including SDS-PAGE banding pattern of storage proteins in seeds of six wild species (Oryza alta, O. grandiglumis, O. meridionalis, O. nivara, O. officinalis and O. rhizomatis) of rice stored under high temperature (45 degrees C) and humidity (approixmately 100%) for 15 days, which facilitated accelerated deterioration. Under the treated conditions, seeds of different wild rice species showed decrease in per cent germination and concentrations of protein and starch, but increase in conductivity of leachate and content of sugar. The SDS-PAGE analysis of seed proteins showed that not only the total number of bands, but also their intensity in terms of thickness differed for each species under storage. The total number of bands ranged from 11 to 22, but none of the species showed all the bands. Similarity index for protein bands between the control and treated seeds was observed to be least in O. rhizomatis and O. alta, while the indices were 0.7 and 0.625 for O. officinalis and O. nivara, respectively. This study clearly showed that seed deterioration led to distinctive biochemical changes, including the presence or absence as well as altered levels of intensity of proteins. Hence, SDS-PAGE protein banding pattern can be used effectively to characterize deterioration of seeds of different wild species of rice.

  1. Quantification of apolipoprotein B-48 and B-100 in rat liver endoplasmic reticulum and Golgi fractions.

    PubMed Central

    Cartwright, I J; Higgins, J A

    1992-01-01

    We have developed a method for measurement of apolipoprotein (apo) B-48 and apo B-100 in blood and subcellular fractions of rat liver based on SDS/PAGE followed by quantitative immunoblotting using 125I-Protein A. Standard curves were prepared in each assay using apo B prepared from total rat lipoproteins by extraction with tetramethylurea. Subcellular fractions (rough and smooth endoplasmic reticulum and Golgi fractions) were prepared from rat liver and separated into membrane and cisternal-content fractions. For quantification, membrane fractions were solubilized in Triton X-100, and the apo B was immunoprecipitated before separation by SDS/PAGE and immunoblotting. Content fractions were concentrated by ultrafiltration and separated by SDS/PAGE without immunoprecipitation. Quantification of apo B in subcellular fractions and detection of apo B by immunoblotting yielded consistent results. In all fractions apo B-48 was the major form, accounting for approximately three-quarters of the total apo B. By using marker enzymes as internal standards, it was calculated that all of the apo B was recovered in the endoplasmic reticulum and Golgi fractions, with approximately 80% of each form of apo B in the endoplasmic reticulum. More than 90% of the apo B of the rough- and smooth-endoplasmic-reticulum fractions was membrane-bound, whereas approx. 33 and 15% of the apo B of the cis-enriched Golgi fractions and trans-enriched Golgi fractions respectively were membrane-bound. Images Fig. 1. Fig. 3. Fig. 4. PMID:1637294

  2. Use of PCR and Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis Techniques for Differentiation of Prevotella intermedia Sensu Stricto and Prevotella nigrescens

    PubMed Central

    Premaraj, Thyagaseely; Kato, Naoki; Fukui, Katsuhito; Kato, Haru; Watanabe, Kunitomo

    1999-01-01

    Primers were designed from 16S rRNA sequences of Prevotella intermedia sensu stricto and Prevotella nigrescens and were used to discriminate these two species by PCR. The results were compared with those from the PCR technique using primers designed from arbitrarily primed PCR products by Guillot and Mouton (E. Guillot and C. Mouton, J. Clin. Microbiol. 35:1876–1882, 1997). The specificities of both assays were studied by using P. intermedia ATCC 25611, P. nigrescens ATCC 33563, 174 clinical isolates of P. intermedia sensu lato, and 59 reference strains and 58 clinical isolates of other Prevotella species and/or common oral flora. In addition, the usefulness and reliability of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in the differentiation of the two species were examined by comparing the results with those from PCR assays. The controversial lipase test for distinguishing these species was also carried out. Unambiguous differentiation was made by both PCR assays, and the results matched each other. The SDS-PAGE assay was found to misidentify a few strains tested, compared with the results of PCR assays. The lipase test was positive for both species, including the reference strains of P. intermedia and P. nigrescens. We conclude that both PCR assays are simple, rapid, reliable, and specific methods which could be used in clinical studies and that the lipase test is not valuable in the differentiation. The reliable discrimination of the two species by SDS-PAGE is questionable. PMID:10074526

  3. Identification of lactic acid bacteria from chili bo, a Malaysian food ingredient.

    PubMed

    Leisner, J J; Pot, B; Christensen, H; Rusul, G; Olsen, J E; Wee, B W; Muhamad, K; Ghazali, H M

    1999-02-01

    Ninety-two strains of lactic acid bacteria (LAB) were isolated from a Malaysian food ingredient, chili bo, stored for up to 25 days at 28 degreesC with no benzoic acid (product A) or with 7,000 mg of benzoic acid kg-1 (product B). The strains were divided into eight groups by traditional phenotypic tests. A total of 43 strains were selected for comparison of their sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) whole-cell protein patterns with a SDS-PAGE database of LAB. Isolates from product A were identified as Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus farciminis, Pediococcus acidilactici, Enterococcus faecalis, and Weissella confusa. Five strains belonging to clusters which could not be allocated to existing species by SDS-PAGE were further identified by 16S rRNA sequence comparison. One strain was distantly related to the Lactobacillus casei/Pediococcus group. Two strains were related to Weissella at the genus or species level. Two other strains did not belong to any previously described 16S rRNA group of LAB and occupied an intermediate position between the L. casei/Pediococcus group and the Weissella group and species of Carnobacterium. The latter two strains belong to the cluster of LAB that predominated in product B. The incidence of new species and subspecies of LAB in chili bo indicate the high probability of isolation of new LAB from certain Southeast Asian foods. None of the isolates exhibited bacteriocin activity against L. plantarum ATCC 14917 and LMG 17682.

  4. Galactinol synthase from kidney bean cotyledon and zucchini leaf. Purification and N-terminal sequences.

    PubMed Central

    Liu, J J; Odegard, W; de Lumen, B O

    1995-01-01

    Galactinol synthase (GS) was purified 1591-fold with a 3.9% recovery from the cotyledon of kidney bean (Phaseolus vulgaris) by a novel scheme consisting of ammonium sulfate fractionation followed by diethylaminoethyl, Affi-Gel Blue, and UDP-hexanolamine affinity chromatography. The purified enzyme had a specific activity of 8.75 mumol mg-1 min-1, a pH optimum of 7.0, and requirements for manganese ion and DTT. The enzyme exhibited a Km = 0.4 mM for UDP-galactose and a Km = 4.5 mM for myo-inositol. It was identified as a 38-kD peptide that co-purified with a 41- and a 43-kD peptide as shown by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Purification to homogeneity was achieved by isolating the 38-kD peptide from the SDS-PAGE gel. To clarify conflicting reports in the literature about the relative molecular mass of purified GS from zucchini leaf (Cucurbita pepo), a similar scheme with modified eluting conditions was used to purify GS from this source. Zucchini leaf GS was purified to homogeneity and identified as a 36-kD peptide on SDS-PAGE. Partial N-terminal sequences of the 38-kD peptide from kidney bean cotyledon and the 36-kD peptide from zucchini leaf were obtained. To facilitate identification of GS during the purification, an assay utilizing thin-layer chromatography and an isotopic analytic imaging scanner was developed. PMID:7480343

  5. Simulation of Ion Motion in FAIMS through Combined Use of SIMION and Modified SDS

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Prasad, Satendra; Tang, Keqi; Manura, David

    2009-11-01

    Over the years, the use of Field Asymmetric Ion Mobility Spectrometry (FAIMS) has grown with applications spanning from explosives detection to separation of complex biological mixtures. Although, the principles of ion separation in FAIMS is understood and comprehensively characterized, little effort has been made in developing commercially available computational tools that can simulate ion motion in FAIMS. Such a tool could be of great value for refining theory, optimizing the performance of the instrument for specific applications, and in modeling the fringe-fields caused by rf decay at the entrance and exit of FAIMS which can significantly affect ion transmission. Anmore » algorithm using SIMIONTM as its core structure was developed in this study to realistically compute ion trajectory at different ratios of electric field to buffer gas number density (E/N). The E/N can vary from a few Td to ~80 Td in FAIMS as created by an asymmetric square waveform. The Statistical Diffusion Simulation (SDS) model was further incorporated in the algorithm to simulate the ion diffusion in the FAIMS gap. The algorithm was validated using a FAIMS analyzer model similar to the Sionex Corporation model SVAC in terms of its dimensions and geometry. Hydroxyproline and Leucine ions with similar reduced mobility Ko (2.17 and 2.18 cm2.V-1.s-1, respectively) were used as model ions to test the new algorithm and demonstrate the effects of gas flow and waveform (voltage pulse amplitude and frequency) on peak shape and ion current transmission. Simulation results from three ion types: O2-(H2O)3, (A type), (C3H6O)2H+ (B type), and (C12H24O)2H+ (C type) were then compared with the experimental data (available in the literature). The SIMION-SDS-Field Dependent Mobility Calculation (FDMC) algorithm provided good agreement with experimental measurements of the ion peak position in FAIMS compensation voltage (CV) spectrum, peak width, and the ion transmission over a broad range of E/N.« less

  6. Preparation of protein samples for mass spectrometry and N-terminal sequencing.

    PubMed

    Glenn, Gary

    2014-01-01

    The preparation of protein samples for mass spectrometry and N-terminal sequencing is a key step in successfully identifying proteins. Mass spectrometry is a very sensitive technique, and as such, samples must be prepared carefully since they can be subject to contamination of the sample (e.g., due to incomplete subcellular fractionation or purification of a multiprotein complex), overwhelming of the sample by highly abundant proteins, and contamination from skin or hair (keratin can be a very common hit). One goal of sample preparation for mass spec is to reduce the complexity of the sample - in the example presented here, mitochondria are purified, solubilized, and fractionated by sucrose density gradient sedimentation prior to preparative 1D SDS-PAGE. It is important to verify the purity and integrity of the sample so that you can have confidence in the hits obtained. More protein is needed for N-terminal sequencing and ideally it should be purified to a single band when run on an SDS-polyacrylamide gel. The example presented here involves stably expressing a tagged protein in HEK293 cells and then isolating the protein by affinity purification and SDS-PAGE. © 2014 Elsevier Inc. All rights reserved.

  7. Analysis of amyloid fibrils in the cheetah (Acinonyx jubatus).

    PubMed

    Bergström, Joakim; Ueda, Mitsuharu; Une, Yumi; Sun, Xuguo; Misumi, Shogo; Shoji, Shozo; Ando, Yukio

    2006-06-01

    Recently, a high prevalence of amyloid A (AA) amyloidosis has been documented among captive cheetahs worldwide. Biochemical analysis of amyloid fibrils extracted from the liver of a Japanese captive cheetah unequivocally showed that protein AA was the main fibril constituent. Further characterization of the AA fibril components by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot analysis revealed three main protein AA bands with approximate molecular weights of 8, 10 and 12 kDa. Mass spectrometry analysis of the 12-kDa component observed in SDS-PAGE and Western blotting confirmed the molecular weight of a 12,381-Da peak. Our finding of a 12-kDa protein AA component provides evidence that the cheetah SAA sequence is longer than the previously reported 90 amino acid residues (approximately 10 kDa), and hence SAA is part of the amyloid fibril.

  8. The impact of Arizona Highways Magazine's facebook page.

    DOT National Transportation Integrated Search

    2014-02-01

    This project examined the relationship between use of the Arizona Highways magazine (AHM) Facebook Page and the decision to : travel to or within Arizona. Key purposes were to: (1) provide a thorough understanding of AHM Facebook Page users, includin...

  9. 47 CFR 22.503 - Paging geographic area authorizations.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... Section 22.503 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES PUBLIC MOBILE SERVICES Paging and Radiotelephone Service § 22.503 Paging geographic area authorizations... in its sole discretion, the FCC determines that the public interest would be served by such...

  10. 47 CFR 22.503 - Paging geographic area authorizations.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... Section 22.503 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES PUBLIC MOBILE SERVICES Paging and Radiotelephone Service § 22.503 Paging geographic area authorizations... in its sole discretion, the FCC determines that the public interest would be served by such...

  11. 47 CFR 22.503 - Paging geographic area authorizations.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... Section 22.503 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES PUBLIC MOBILE SERVICES Paging and Radiotelephone Service § 22.503 Paging geographic area authorizations... in its sole discretion, the FCC determines that the public interest would be served by such...

  12. Required Discussion Web Pages in Psychology Courses and Student Outcomes

    ERIC Educational Resources Information Center

    Pettijohn, Terry F., II; Pettijohn, Terry F.

    2007-01-01

    We conducted 2 studies that investigated student outcomes when using discussion Web pages in psychology classes. In Study 1, we assigned 213 students enrolled in Introduction to Psychology courses to either a mandatory or an optional Web page discussion condition. Students used the discussion Web page significantly more often and performed…

  13. Beta-Carotene chemical stability in nanoemulsions was improved by stabilized with Beta-Lactoglobulin-Catechin conjugates through free radical method

    USDA-ARS?s Scientific Manuscript database

    Beta-lactoglobulin (BLG)-catechin conjugates were prepared by a free radical method and investigated with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), electrospray ionization-mass spectrometry (ESI-MS), and far-UV circular dichroism (CD). Covalent binding between BLG and cat...

  14. Glycation inhibits trichloroacetic acid (TCA)-induced whey protein precipitation

    USDA-ARS?s Scientific Manuscript database

    Four different WPI saccharide conjugates were successfully prepared to test whether glycation could inhibit WPI precipitation induced by trichloroacetic acid (TCA). Conjugates molecular weights after glycation were analyzed with SDS-PAGE. No significant secondary structure change due to glycation wa...

  15. PURIFICATION AND PARTIAL CHARACTERIZATION OF AN ACID PHOSPHATASE FROM SPIRODELA OLIGORRHIZA AND ITS AFFINITY FOR SELECTED ORGANOPHOSPHATE PESTICIDES

    EPA Science Inventory

    An acid phosphatase from the aquatic plant Spirodela oligorrhiza (duckweed) was isolated by fast protein liquid chromatography (FPLC) and partially characterized. The enzyme was purified 1871-fold with a total yield of 40%. SDS-PAGE electrophoresis of the pure acid phosphatase ...

  16. SPAX - PAX with Super-Pages

    NASA Astrophysics Data System (ADS)

    Bößwetter, Daniel

    Much has been written about the pros and cons of column-orientation as a means to speed up read-mostly analytic workloads in relational databases. In this paper we try to dissect the primitive mechanisms of a database that help express the coherence of tuples and present a novel way of organizing relational data in order to exploit the advantages of both, the row-oriented and the column-oriented world. As we go, we break with yet another bad habit of databases, namely the equal granularity of reads and writes which leads us to the introduction of consecutive clusters of disk pages called super-pages.

  17. The conformational state of polyphenol oxidase from field bean (Dolichos lablab) upon SDS and acid-pH activation

    PubMed Central

    Kanade, Santosh R.; Paul, Beena; Rao, A. G. Appu; Gowda, Lalitha R.

    2006-01-01

    Field bean (Dolichos lablab) contains a single isoform of PPO (polyphenol oxidase) – a type III copper protein that catalyses the o-hydroxylation of monophenols and oxidation of o-diphenols using molecular oxygen – and is a homotetramer with a molecular mass of 120 kDa. The enzyme is activated manyfold either in the presence of the anionic detergent SDS below its critical micellar concentration or on exposure to acid-pH. The enhancement of kcat upon activation is accompanied by a marked shift in the pH optimum for the oxidation of t-butyl catechol from 4.5 to 6.0, an increased sensitivity to tropolone, altered susceptibility to proteolytic degradation and decreased thermostability. The Stokes radius of the native enzyme is found to increase from 49.1±2 to 75.9±0.6 Å (1 Å=0.1 nm). The activation by SDS and acid-pH results in a localized conformational change that is anchored around the catalytic site of PPO that alters the microenvironment of an essential glutamic residue. Chemical modification of field bean and sweet potato PPO with 1-ethyl-3-(3-dimethylaminopropyl)carbodi-imide followed by kinetic analysis leads to the conclusion that both the enzymes possess a core carboxylate essential to activity. This enhanced catalytic efficiency of PPO, considered as an inducible defence oxidative enzyme, is vital to the physiological defence strategy adapted by plants to insect herbivory and pathogen attack. PMID:16393141

  18. Making Pages That Move.

    ERIC Educational Resources Information Center

    Gepner, Ivan

    2001-01-01

    Explains the mechanism of producing dynamic computer pages which is based on three technologies: (1) the document object model; (2) cascading stylesheets; and (3) javascript. Discusses the applications of these techniques in genetics and developmental biology. (YDS)

  19. Heap/stack guard pages using a wakeup unit

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gooding, Thomas M; Satterfield, David L; Steinmacher-Burow, Burkhard

    A method and system for providing a memory access check on a processor including the steps of detecting accesses to a memory device including level-1 cache using a wakeup unit. The method includes invalidating level-1 cache ranges corresponding to a guard page, and configuring a plurality of wakeup address compare (WAC) registers to allow access to selected WAC registers. The method selects one of the plurality of WAC registers, and sets up a WAC register related to the guard page. The method configures the wakeup unit to interrupt on access of the selected WAC register. The method detects access ofmore » the memory device using the wakeup unit when a guard page is violated. The method generates an interrupt to the core using the wakeup unit, and determines the source of the interrupt. The method detects the activated WAC registers assigned to the violated guard page, and initiates a response.« less

  20. Automatic Hidden-Web Table Interpretation by Sibling Page Comparison

    NASA Astrophysics Data System (ADS)

    Tao, Cui; Embley, David W.

    The longstanding problem of automatic table interpretation still illudes us. Its solution would not only be an aid to table processing applications such as large volume table conversion, but would also be an aid in solving related problems such as information extraction and semi-structured data management. In this paper, we offer a conceptual modeling solution for the common special case in which so-called sibling pages are available. The sibling pages we consider are pages on the hidden web, commonly generated from underlying databases. We compare them to identify and connect nonvarying components (category labels) and varying components (data values). We tested our solution using more than 2,000 tables in source pages from three different domains—car advertisements, molecular biology, and geopolitical information. Experimental results show that the system can successfully identify sibling tables, generate structure patterns, interpret tables using the generated patterns, and automatically adjust the structure patterns, if necessary, as it processes a sequence of hidden-web pages. For these activities, the system was able to achieve an overall F-measure of 94.5%.

  1. Multiplex PageRank.

    PubMed

    Halu, Arda; Mondragón, Raúl J; Panzarasa, Pietro; Bianconi, Ginestra

    2013-01-01

    Many complex systems can be described as multiplex networks in which the same nodes can interact with one another in different layers, thus forming a set of interacting and co-evolving networks. Examples of such multiplex systems are social networks where people are involved in different types of relationships and interact through various forms of communication media. The ranking of nodes in multiplex networks is one of the most pressing and challenging tasks that research on complex networks is currently facing. When pairs of nodes can be connected through multiple links and in multiple layers, the ranking of nodes should necessarily reflect the importance of nodes in one layer as well as their importance in other interdependent layers. In this paper, we draw on the idea of biased random walks to define the Multiplex PageRank centrality measure in which the effects of the interplay between networks on the centrality of nodes are directly taken into account. In particular, depending on the intensity of the interaction between layers, we define the Additive, Multiplicative, Combined, and Neutral versions of Multiplex PageRank, and show how each version reflects the extent to which the importance of a node in one layer affects the importance the node can gain in another layer. We discuss these measures and apply them to an online multiplex social network. Findings indicate that taking the multiplex nature of the network into account helps uncover the emergence of rankings of nodes that differ from the rankings obtained from one single layer. Results provide support in favor of the salience of multiplex centrality measures, like Multiplex PageRank, for assessing the prominence of nodes embedded in multiple interacting networks, and for shedding a new light on structural properties that would otherwise remain undetected if each of the interacting networks were analyzed in isolation.

  2. TVDG Home Page

    Science.gov Websites

    Download WinZip Radiation Effects Testing and Calibration This facility is available for the study of space radiation effects, in particular, Single Event Upset ( SEU ) Testing and Spacecraft Instrument Calibration information about our facility. Visit our Space and Radiation Effects Links page to find out what is going on

  3. SdsA polymorph isolation and improvement of their crystal quality using nonconventional crystallization techniques

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    De la Mora, Eugenio; Flores-Hernández, Edith; Jakoncic, Jean

    SdsA, a sodium dodecyl sulfate hydrolase, from Pseudomonas aeruginosa was crystallized in three different crystal polymorphs and their three-dimensional structure was determined. The different polymorphs present different crystal packing habits. One of the polymorphs suggests the existence of a tetramer, an oligomeric state not observed previously, while the crystal packing of the remaining two polymorphs obstructs the active site entrance but stabilizes flexible regions of the protein. Nonconventional crystallization methods that minimize convection, such as counterdiffusion in polyvinyl alcohol gel coupled with the influence of a 500 MHz (10.2 T) magnetic field, were necessary to isolate the poorest diffracting polymorphmore » and increase its internal order to determine its structure by X-ray diffraction. In conclusion, the results obtained show the effectiveness of nonconventional crystallographic methods to isolate different crystal polymorphs.« less

  4. World Wide Web Page Design: A Structured Approach.

    ERIC Educational Resources Information Center

    Gregory, Gwen; Brown, M. Marlo

    1997-01-01

    Describes how to develop a World Wide Web site based on structured programming concepts. Highlights include flowcharting, first page design, evaluation, page titles, documenting source code, text, graphics, and browsers. Includes a template for HTML writers, tips for using graphics, a sample homepage, guidelines for authoring structured HTML, and…

  5. The Library as Information Provider: The Home Page.

    ERIC Educational Resources Information Center

    Clyde, Laurel A.

    1996-01-01

    Discusses ways in which libraries are using the World Wide Web to provide information via a home page, based on information from a survey in Iceland as well as a larger study that conducted content analyses of home pages of public and school libraries in 13 countries. (Author/LRW)

  6. Dynamic Web Pages: Performance Impact on Web Servers.

    ERIC Educational Resources Information Center

    Kothari, Bhupesh; Claypool, Mark

    2001-01-01

    Discussion of Web servers and requests for dynamic pages focuses on experimentally measuring and analyzing the performance of the three dynamic Web page generation technologies: CGI, FastCGI, and Servlets. Develops a multivariate linear regression model and predicts Web server performance under some typical dynamic requests. (Author/LRW)

  7. The changes of serum proteome and tissular pathology in mouse induced by botulinum toxin E injection.

    PubMed

    Wang, J F; Mao, X Y; Zhao, C

    2014-01-01

    The experiment were performed to investigate the poisoning-related proteins and main pathological changes after mouse suffered from injection of botulinum toxin serotype E. Dose of 0.75 LD50 botulinum toxin serotype E per mice were administrated by intraperitoneal injection. Survival mouse were picked as experimental group. The blood were collected from orbital blood and serum sample was separated by centrifugation. The heart, liver, spleen, lung, kidney were fixed in 10 % neutral buffered formalin and then developed paraffin sections. Serum protein components were analyzed by SDS-PAGE gel electrophoresis coupled with 2-DE SDS-PAGE gel electrophoresis. Differentially expressed proteins were analyzed by PDQUest8.0 software and subjected to ion trap mass spectrometry equipped with a high performance liquid chromatography system. The observation of pathological section showed that heart, liver, spleen, lung, kidney exhibited pathological changes in different degree, especially in heart, liver and lung tissues. Heart muscle tissue display serious inflammatory response, heart muscle fiber compulsively expanded and filled with erythrocyte and inflammatory exudates, some heart muscle fiber ruptured, even necrosis; hepatic cell in edge of liver occur apoptosis and some hepatic cell have disintegrated, and even died; pulmonary alveoli broken and partial vein filled with blood. Serum proteins component present a significant changes between control serum and botulism in 24 h by SDS-PAGE gel electrophoresis and 2-DE-SDS-PAGE gel electrophoresis. Twenty differentially expressed protein spots were observed in 2-DE profiles, in which 14 protein spots were undetectable in serum proteome under botulism, 3 protein spots exclusively expressed in state of botulism, 3 protein spots were low-expressed in serum proteome under botulism. Fourteen proteins have been identified among 20 spots elected on two-dimensional electrophoresis gels. Crystal proteins family exclusively expressed in

  8. The mediating role of facebook fan pages.

    PubMed

    Chih, Wen-Hai; Hsu, Li-Chun; Wang, Kai-Yu; Lin, Kuan-Yu

    2014-01-01

    Using the dual mediation hypothesis, this study investigates the role of interestingness (the power of attracting or holding one's attention) attitude towards the news, in the formation of Facebook Fan Page users' electronic word-of-mouth intentions. A total of 599 Facebook fan page users in Taiwan were recruited and structural equation modeling (SEM) was used to test the research hypotheses. The results show that both perceived news entertainment and informativeness positively influence interestingness attitude towards the news. Interestingness attitude towards the news subsequently influences hedonism and utilitarianism attitudes towards the Fan Page, which then influence eWOM intentions. Interestingness attitude towards the news plays a more important role than hedonism and utilitarianism attitudes in generating electronic word-of-mouth intentions. Based on the findings, the implications and future research suggestions are provided.

  9. Identification of a low digestibility δ-Conglutin in yellow lupin (Lupinus luteus L.) seed meal for atlantic salmon (Salmo salar L.) by coupling 2D-PAGE and mass spectrometry.

    PubMed

    Ogura, Takahiro; Hernández, Adrián; Aizawa, Tomoko; Ogihara, Jun; Sunairi, Michio; Alcaino, Javier; Salvo-Garrido, Haroldo; Maureira-Butler, Iván J

    2013-01-01

    The need of quality protein in the aquaculture sector has forced the incorporation of alternative plant proteins into feeding diets. However, most plant proteins show lower digestibility levels than fish meal proteins, especially in carnivorous fishes. Manipulation of protein content by plant breeding can improve the digestibility rate of plant proteins in fish, but the identification of low digestibility proteins is essential. A reduction of low digestibility proteins will not only increase feed efficiency, but also reduce water pollution. Little is known about specific digestible protein profiles and/or molecular identification of more bioavailable plant proteins in fish diets. In this study, we identified low digestibility L. luteus seed proteins using Atlantic salmon (Salmo salar) crude digestive enzymes in an in vitro assay. Low digestibility proteins were identified by comparing SDS-PAGE banding profiles of digested and non-digested lupin seed proteins. Gel image analysis detected a major 12 kDa protein band in both lupin meal and protein isolate digested products. The 12 kDa was confirmed by 2D-PAGE gels and the extracted protein was analyzed with an ion trap mass spectrometer in tandem mass mode. The MS/MS data showed that the 12 kDa low digestibility protein was a large chain δconglutin, a common seed storage protein of yellow lupin. Comparison of the protein band profiles between lupin meal and protein isolates showed that the isolatation process did not affect the low digestibility of the 12 kDa protein.

  10. Identification of a Low Digestibility δ-Conglutin in Yellow Lupin (Lupinus luteus L.) Seed Meal for Atlantic Salmon (Salmo salar L.) by Coupling 2D-PAGE and Mass Spectrometry

    PubMed Central

    Ogura, Takahiro; Hernández, Adrián; Aizawa, Tomoko; Ogihara, Jun; Sunairi, Michio; Alcaino, Javier; Salvo-Garrido, Haroldo; Maureira-Butler, Iván J.

    2013-01-01

    The need of quality protein in the aquaculture sector has forced the incorporation of alternative plant proteins into feeding diets. However, most plant proteins show lower digestibility levels than fish meal proteins, especially in carnivorous fishes. Manipulation of protein content by plant breeding can improve the digestibility rate of plant proteins in fish, but the identification of low digestibility proteins is essential. A reduction of low digestibility proteins will not only increase feed efficiency, but also reduce water pollution. Little is known about specific digestible protein profiles and/or molecular identification of more bioavailable plant proteins in fish diets. In this study, we identified low digestibility L. luteus seed proteins using Atlantic salmon (Salmo salar) crude digestive enzymes in an in vitro assay. Low digestibility proteins were identified by comparing SDS-PAGE banding profiles of digested and non-digested lupin seed proteins. Gel image analysis detected a major 12 kDa protein band in both lupin meal and protein isolate digested products. The 12 kDa was confirmed by 2D-PAGE gels and the extracted protein was analyzed with an ion trap mass spectrometer in tandem mass mode. The MS/MS data showed that the 12 kDa low digestibility protein was a large chain δconglutin, a common seed storage protein of yellow lupin. Comparison of the protein band profiles between lupin meal and protein isolates showed that the isolatation process did not affect the low digestibility of the 12 kDa protein. PMID:24278278

  11. Pristine multi-walled carbon nanotubes/SDS modified carbon paste electrode as an amperometric sensor for epinephrine.

    PubMed

    Thomas, Tony; Mascarenhas, Ronald J; D' Souza, Ozma J; Detriche, Simon; Mekhalif, Zineb; Martis, Praveen

    2014-07-01

    An amperometric sensor for the determination of epinephrine (EP) was fabricated by modifying the carbon paste electrode (CPE) with pristine multi-walled carbon nanotubes (pMWCNTs) using bulk modification followed by drop casting of sodium dodecyl sulfate (SDS) onto the surface for its optimal potential application. The modified electrode showed an excellent electrocatalytic activity towards EP by decreasing the overpotential and greatly enhancing the current sensitivity. FE-SEM images confirmed the dispersion of pMWCNTs in the CPE matrix. EDX analysis ensured the surface coverage of SDS. A comparative study of pMWCNTs with those of oxidized MWCNTs (MWCNTsOX) modified electrodes reveals that the former is the best base material for the construction of the sensor with advantages of lower oxidation overpotential and the least background current. The performance of the modified electrode was impressive in terms of the least charge transfer resistance (Rct), highest values for diffusion coefficient (DEP) and standard heterogeneous electron transfer rate constant (k°). Analytical characterization of the modified electrode exhibited two linear dynamic ranges from 1.0×10(-7) to 1.0×10(-6)M and 1.0×10(-6) to 1.0×10(-4)M with a detection limit of (4.5±0.18)×10(-8)M. A 100-fold excess of serotonin, acetaminophen, folic acid, uric acid, tryptophan, tyrosine and cysteine, 10-fold excess of ascorbic acid and twofold excess of dopamine do not interfere in the quantification of EP at this electrode. The analytical applications of the modified electrode were demonstrated by determining EP in spiked blood serum and adrenaline tartrate injection. The modified electrode involves a simple fabrication procedure, minimum usage of the modifier, quick response, excellent stability, reproducibility and anti-fouling effects. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Modification of an acetone-sodium dodecyl sulfate disruption method for cellular protein extraction from neuropathogenic Clostridium botulinum

    USDA-ARS?s Scientific Manuscript database

    An acetone-sodium dodecyl sulfate (SDS) disruption method was used for the extraction of cellular proteins from neurotoxigenic Clostridium botulinum. The amount of protein extracted per gram of dry weight and the protein profile as revealed by polyacrylamide gel electrophoresis (PAGE) was comparabl...

  13. Oncoprotein protein kinase

    DOEpatents

    Karin, Michael; Hibi, Masahiko; Linn, Anning

    1996-01-01

    An isolated polypeptide (JNK) characterized by having a molecular weight of 46kD as determined by reducing SDS-PAGE, having serine and threonine kinase activity, phosphorylating the c-Jun N-terminal activation domain and polynucleotide sequences and method of detection of JNK.

  14. Portable exhausters POR-004 SKID B, POR-005 SKID C, POR-006 SKID D storage plan

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Nelson, O.D.

    1997-09-04

    This document provides a storage plan for portable exhausters POR-004 SKID B, POR-005 SKID C, AND POR-006 SKID D. The exhausters will be stored until they are needed by the TWRS (Tank Waste Remediation Systems) Saltwell Pumping Program. The storage plan provides criteria for portable exhauster storage, periodic inspections during storage, and retrieval from storage.

  15. Attention: Page has moved

    Science.gov Websites

    Weather Service NWS logo - Click to go to the NWS home page Climate Prediction Center Site Map News Prediction Climate Prediction Center 5200 Auth Road Camp Springs, Maryland 20746 Climate Prediction Center

  16. The proteins of the grape (Vitis vinifera L.) seed endosperm: fractionation and identification of the major components.

    PubMed

    Gazzola, Diana; Vincenzi, Simone; Gastaldon, Luca; Tolin, Serena; Pasini, Gabriella; Curioni, Andrea

    2014-07-15

    In the present study, grape (Vitis vinifera L.) seed endosperm proteins were characterized after sequential fractionation, according to a modified Osborne procedure. The salt-soluble fraction (albumins and globulins) comprised the majority (58.4%) of the total extracted protein. The protein fractions analysed by SDS-PAGE showed similar bands, indicating different solubility of the same protein components. SDS-PAGE in non-reducing and reducing conditions revealed the polypeptide composition of the protein bands. The main polypeptides, which were similar in all the grape varieties analysed, were identified by LC-MS/MS as homologous to the 11S globulin-like seed storage proteins of other plant species, while a monomeric 43 kDa protein presented high homology with the 7S globulins of legume seeds. The results provide new insights about the identity, structure and polypeptide composition of the grape seed storage proteins. Copyright © 2014 Elsevier Ltd. All rights reserved.

  17. Development of a wheat-Aegilops searsii substitution line with positively affecting Chinese steamed bread quality

    PubMed Central

    Du, Xuye; Ma, Xin; Min, Jingzhi; Zhang, Xiaocun; Jia, Zhenzhen

    2018-01-01

    A wheat-Aegilops searsii substitution line GL1402, in which chromosome 1B was substituted with 1Ss from Ae. searsii, was developed and detected using SDS-PAGE and GISH. The SDS-PAGE analysis showed that the HMW-GS encoded by the Glu-B1 loci of Chinese Spring was replaced by the HMW-GS encoded by the Glu-1Ss loci of Ae. searsii. Glutenin macropolymer (GMP) investigation showed that GL1402 had a much higher GMP content than Chinese Spring did. A dough quality comparison of GL1402 and Chinese Spring indicated that GL1402 showed a significantly higher protein content and middle peak time (MPT), and a smaller right peak slope (RPS). Quality tests of Chinese steamed bread (CSB) showed that the GL1402 also produced good steamed bread quality. These results suggested that the substitution line is a valuable breeding material for improving the wheat processing quality.

  18. BRCA1 Regulation of Estrogen Signaling in the Breast

    DTIC Science & Technology

    2007-05-01

    PAGE and processed by immunoblot analysis with antibodies specific for GAL4- DBD or ER as indicated by arrows. Note that differences in the relative...derivative 1–150 (N terminus). Schematic diagrams of ER and DBC-1 indicate fragments used in binding reactions. AF-1, Activation function 1; DBD , DNA...solved by SDS-10% PAGE. Proteins were analyzed by im- munoblot using antibodies against GAL4- DBD (RK5C1; Santa Cruz Biotechnology, Santa Cruz, CA

  19. Four Single-Page Learning Models.

    ERIC Educational Resources Information Center

    Hlynka, Denis

    1979-01-01

    Identifies four models of single-page learning systems that can streamline lengthy, complex prose: Information Mapping, Focal Press Model, Behavioral Objectives Model, and School Mathematics Model. (CMV)

  20. Toward a User-Centered Academic Library Home Page

    ERIC Educational Resources Information Center

    McHale, Nina

    2008-01-01

    In the past decade, academic libraries have struggled with the design of an effective library home page. Since librarians' mental models of information architecture differ from those of their patrons, usability assessments are necessary in designing a user-centered home page. This study details a usability sequence of card sort and paper and…

  1. How Egg Case Proteins Can Protect Cuttlefish Offspring?

    PubMed Central

    Cornet, Valérie; Henry, Joël; Goux, Didier; Duval, Emilie; Bernay, Benoit; Le Corguillé, Gildas; Corre, Erwan; Zatylny-Gaudin, Céline

    2015-01-01

    Sepia officinalis egg protection is ensured by a complex capsule produced by the female accessory genital glands and the ink bag. Our study is focused on the proteins constituting the main egg case. De novo transcriptomes from female genital glands provided essential databases for protein identification. A proteomic approach in SDS-PAGE coupled with MS unveiled a new egg case protein family: SepECPs, for Sepia officinalis Egg Case Proteins. N-glycosylation was demonstrated by PAS staining SDS-PAGE gels. These glycoproteins are mainly produced in the main nidamental glands. SepECPs share high sequence homology, especially in the signal peptide and the three cysteine-rich domains. SepECPs have a high number of cysteines, with conserved motifs involved in 3D-structure. SDS-PAGE showed that SepECPs could form dimers; this result was confirmed by TEM observations, which also revealed a protein network. This network is similar to the capsule network, and it associates these structural proteins with polysaccharides, melanin and bacteria to form a tight mesh. Its hardness and elasticity provide physical protection to the embryo. In addition, SepECPs also have bacteriostatic antimicrobial activity on GRAM- bacteria. By observing the SepECP / Vibrio aestuarianus complex in SEM, we demonstrated the ability of these proteins to agglomerate bacteria and thus inhibit their growth. These original proteins identified from the outer egg case ensure the survival of the species by providing physical and chemical protection to the embryos released in the environment without any maternal protection. PMID:26168161

  2. Diagnostic potential of low molecular weight excretory secretory proteins of Paramphistomum epiclitum for caprine amphistomosis.

    PubMed

    Jaiswal, Amit Kumar; Shanker, Daya; Sudan, Vikrant; Singh, Amit

    2018-06-15

    In the present study, the 75% alcoholic fractionation of excretory-secretory (ES) antigen isolated from 200 to 300 live P. epiclitum was assessed for its diagnostic potential for the detection of caprine amphistomosis by using antibody detection enzyme immunoassay. Prior to enzyme immunoassay, 75% alcoholic fractionation of excretory-secretory (ES) antigen was subjected to SDS- PAGE and western blot analysis for the presence of immunoreactive polypeptides. SDS-PAGE analysis of ES antigen resolved a total 7 polypeptides bands of size 56, 27, 25, 22.5, 12, 11 and 10 kDa. Western blot analysis revealed only two immunoreactive polypeptides (11 kDa and 12 kDa) when polypeptides resolved in SDS-PAGE were probed with known positive pooled serum. None of the polypeptides showed reactions with pooled known negative serum. The working dilutions of antigen, sera and conjugates were determined by checkerboard titration for employing ELISA and cut-off O.D. was calculated 0.616 by using the mean absorbance value of 11 negative kid sera. The sensitivity and specificity of ELISA was found to be 100% and 86.76%, respectively. As per kappa value estimation, the strength of agreement was found to be good. Antibodies to 75% alcoholic fractionation of ES antigen was detected in 20% goats (n = 160) of either sex, although faecal examination detected 10.6% goats to be infected with amphistomosis. The study confirmed that 75% alcoholic fractionation of ES antigen of P. epiclitum based ELISA had good value for serodiagnosis of caprine amphistomosis. Copyright © 2018 Elsevier B.V. All rights reserved.

  3. Identification of Lactic Acid Bacteria from Chili Bo, a Malaysian Food Ingredient

    PubMed Central

    Leisner, Jørgen J.; Pot, Bruno; Christensen, Henrik; Rusul, Gulam; Olsen, John E.; Wee, Bee Wah; Muhamad, Kharidah; Ghazali, Hasanah M.

    1999-01-01

    Ninety-two strains of lactic acid bacteria (LAB) were isolated from a Malaysian food ingredient, chili bo, stored for up to 25 days at 28°C with no benzoic acid (product A) or with 7,000 mg of benzoic acid kg−1 (product B). The strains were divided into eight groups by traditional phenotypic tests. A total of 43 strains were selected for comparison of their sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) whole-cell protein patterns with a SDS-PAGE database of LAB. Isolates from product A were identified as Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus farciminis, Pediococcus acidilactici, Enterococcus faecalis, and Weissella confusa. Five strains belonging to clusters which could not be allocated to existing species by SDS-PAGE were further identified by 16S rRNA sequence comparison. One strain was distantly related to the Lactobacillus casei/Pediococcus group. Two strains were related to Weissella at the genus or species level. Two other strains did not belong to any previously described 16S rRNA group of LAB and occupied an intermediate position between the L. casei/Pediococcus group and the Weissella group and species of Carnobacterium. The latter two strains belong to the cluster of LAB that predominated in product B. The incidence of new species and subspecies of LAB in chili bo indicate the high probability of isolation of new LAB from certain Southeast Asian foods. None of the isolates exhibited bacteriocin activity against L. plantarum ATCC 14917 and LMG 17682. PMID:9925588

  4. Production and purification of polyclonal antibody against F(ab')2 fragment of human immunoglobulin G

    PubMed Central

    Nasiri, Hadi; Valedkarimi, Zahra; Aghebati-Maleki, Leili; Abdolalizadeh, Jalal; Kazemi, Tohid; Esparvarinha, Mojghan; Majidi, Jafar

    2017-01-01

    Antibodies are essential tools of biomedical and biochemical researches. Polyclonal antibodies are produced against different epitopes of antigens. Purified F(ab')2 can be used for animal’s immunization to produce polyclonal antibodies. Human immunoglobulin G (IgG) was purified by ion exchange chromatography method. In all stages verification method of the purified antibodies was sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Purified IgG was digested by pepsin enzyme and F(ab')2 fragment was purified by gel filtration separation method. For production of polyclonal antibody, rabbit was immunized by purified F(ab')2 and antibody production was investigated by enzyme-linked immunosorbent assay. Purified anti-IgG F(ab')2 was conjugated with fluorescein isothiocyanate. Ion exchange chromatography purification yielded 38 mg of human IgG antibody. The results of SDS-PAGE in reduced and non-reduced conditions showed bands with 25-30 kDa molecular weight (MW) and 50-kDa respectively and a distinct band with 150 kDa MW. The results of non-reduced SDS-PAGE for determining the purity of F(ab')2 fragment showed one band in 90 kDa and a band in 150 kDa MW position. Purification by Ion exchange chromatography method resulted about 12 mg rabbit polyclonal antibody. Flow cytometry showed generated polyclonal antibody had an acceptable activity compared to commercial antibody. Taking together, purified IgG F(ab')2 and polyclonal anti-IgG F(ab')2 are useful tools in biomedical and biochemical researches and diagnostic kits. PMID:29326789

  5. Production and purification of polyclonal antibody against F(ab')2 fragment of human immunoglobulin G.

    PubMed

    Nasiri, Hadi; Valedkarimi, Zahra; Aghebati-Maleki, Leili; Abdolalizadeh, Jalal; Kazemi, Tohid; Esparvarinha, Mojghan; Majidi, Jafar

    2017-01-01

    Antibodies are essential tools of biomedical and biochemical researches. Polyclonal antibodies are produced against different epitopes of antigens. Purified F(ab') 2 can be used for animal's immunization to produce polyclonal antibodies. Human immunoglobulin G (IgG) was purified by ion exchange chromatography method. In all stages verification method of the purified antibodies was sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Purified IgG was digested by pepsin enzyme and F(ab') 2 fragment was purified by gel filtration separation method. For production of polyclonal antibody, rabbit was immunized by purified F(ab') 2 and antibody production was investigated by enzyme-linked immunosorbent assay. Purified anti-IgG F(ab') 2 was conjugated with fluorescein isothiocyanate. Ion exchange chromatography purification yielded 38 mg of human IgG antibody. The results of SDS-PAGE in reduced and non-reduced conditions showed bands with 25-30 kDa molecular weight (MW) and 50-kDa respectively and a distinct band with 150 kDa MW. The results of non-reduced SDS-PAGE for determining the purity of F(ab') 2 fragment showed one band in 90 kDa and a band in 150 kDa MW position. Purification by Ion exchange chromatography method resulted about 12 mg rabbit polyclonal antibody. Flow cytometry showed generated polyclonal antibody had an acceptable activity compared to commercial antibody. Taking together, purified IgG F(ab') 2 and polyclonal anti-IgG F(ab') 2 are useful tools in biomedical and biochemical researches and diagnostic kits.

  6. House-dust mite allergy: mapping of Dermatophagoides pteronyssinus allergens for dogs by two-dimensional immunoblotting.

    PubMed

    Martins, Luís Miguel Lourenço; Marques, Andreia Grilo; Pereira, Luísa Maria Dotti Silva; Goicoa, Ana; Semião-Santos, Saul José; Bento, Ofélia Pereira

    2015-04-01

    Specific immunotherapy has shown to be very useful for allergy control in dogs, with a common success rate ranging from 65% to 70%. However, this efficacy could probably be improved and the identification of individual allergomes, with the choice of more adequate molecular allergen pools for specific immunotherapy, being the strategy. To map Dermatophagoides pteronyssinus (Der p) allergens for mite-sensitized atopic dogs, for better understanding how individual allergograms may influence the response to house-dust mite immunotherapy. To identify the Der p mite allergome for dogs, 20 individuals allergic to dust-mites and sensitized to Der p, were selected. The extract from Der p was submitted to isoelectric focusing (IEF), one-dimensional (1-D) and two-dimensional (2-D) sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Separated proteins were blotted onto polyvinylidene difluoride (PVDF) membranes and immunoblottings were performed with patient sera. Allergen-bound specific IgE was detected. Eleven allergens were identified from isoelectric focusing (IEF), as well as from 1-D SDS PAGE. From 2-D SDS-PAGE, 24 spots were identified. Several similarities were found between dog and human allergograms and no absolute correlation between sensitization and allergy was observed either. As in humans, different individual allergograms do not seem to implicate different clinical patterns, but may influence the response to specific immunotherapy. The molecular epidemiology approach in veterinary allergy management, by the characterization of individual patients' allergoms and by choosing the best molecular allergen pool for each patient could also improve the efficacy of allergy immunotherapy.

  7. Properties of dermonecrotic toxin prepared from sonic extracts Bordetella bronchiseptica.

    PubMed

    Kume, K; Nakai, T; Samejima, Y; Sugimoto, C

    1986-05-01

    A toxin with dermonecrotic activity (DNT) was purified from sonic extracts of Bordetella bronchiseptica L3 of pig origin at phase I by chromatographic and electrophoretic methods. The purification procedure was one developed for obtaining the Pasteurella multocida DNT from sonic extracts with some modifications. Dermonecrotizing activity of B. bronchiseptica-purified DNT was increased by 600-fold compared with that of the crude extract, and the average yield was about 3%. The toxin was homogeneous, as determined by Ouchterlony double immunodiffusion, crossed immunoelectrophoresis, and disk isoelectric focusing in polyacrylamide gels. The toxin gave a single band on polyacrylamide disk gel electrophoresis (PAGE) and sodium dodecyl sulfate-SDS PAGE. The molecular weight of the toxin was ca. 190,000 +/- 5,000, as determined by SDS-PAGE. The isoelectric point of the toxin was ca. 6.5 to 6.6. The minimal necrotizing dose of the toxin for guinea pigs was about 2 ng of protein per 0.1 ml, the 50% lethal dose per mouse was about 0.3 micrograms, and the minimal cytotoxic dose for embryonic bovine lung cells was about 2 ng/ml. The toxin was heat labile and sensitive to inactivation by trypsin, Formalin, and glutaraldehyde. The mildly trypsinized B. bronchiseptica DNT preparation dissociated into two polypeptide chains, with molecular weights of ca. 75,000 +/- 4,000 (fragment 1) and ca. 118,000 +/- 5,000 (fragment 2), after treatment with dithiothreitol-SDS or urea. Upon removal of dithiothreitol and urea from the dissociated DNT preparation, the fragments reassociated, and the DNT that was formed was indistinguishable from the native toxin.

  8. Handley Page metal construction

    NASA Technical Reports Server (NTRS)

    1929-01-01

    In this report Handley Page construction techniques are shown such as: solid-drawn tubular duralumin spars are used in the stabilizer; plain channel sections are used extensively for minor components; and the manner of assembling them into a stabilizer compression strut is shown.

  9. Poisson statistics of PageRank probabilities of Twitter and Wikipedia networks

    NASA Astrophysics Data System (ADS)

    Frahm, Klaus M.; Shepelyansky, Dima L.

    2014-04-01

    We use the methods of quantum chaos and Random Matrix Theory for analysis of statistical fluctuations of PageRank probabilities in directed networks. In this approach the effective energy levels are given by a logarithm of PageRank probability at a given node. After the standard energy level unfolding procedure we establish that the nearest spacing distribution of PageRank probabilities is described by the Poisson law typical for integrable quantum systems. Our studies are done for the Twitter network and three networks of Wikipedia editions in English, French and German. We argue that due to absence of level repulsion the PageRank order of nearby nodes can be easily interchanged. The obtained Poisson law implies that the nearby PageRank probabilities fluctuate as random independent variables.

  10. Automatic comic page image understanding based on edge segment analysis

    NASA Astrophysics Data System (ADS)

    Liu, Dong; Wang, Yongtao; Tang, Zhi; Li, Luyuan; Gao, Liangcai

    2013-12-01

    Comic page image understanding aims to analyse the layout of the comic page images by detecting the storyboards and identifying the reading order automatically. It is the key technique to produce the digital comic documents suitable for reading on mobile devices. In this paper, we propose a novel comic page image understanding method based on edge segment analysis. First, we propose an efficient edge point chaining method to extract Canny edge segments (i.e., contiguous chains of Canny edge points) from the input comic page image; second, we propose a top-down scheme to detect line segments within each obtained edge segment; third, we develop a novel method to detect the storyboards by selecting the border lines and further identify the reading order of these storyboards. The proposed method is performed on a data set consisting of 2000 comic page images from ten printed comic series. The experimental results demonstrate that the proposed method achieves satisfactory results on different comics and outperforms the existing methods.

  11. WMO SDS-WAS NAMEE Regional Center: Towards continuous evaluation of dust models in Northern Africa

    NASA Astrophysics Data System (ADS)

    Basart, Sara; García-Castillo, Gerardo; Cuevas, Emilio; Terradellas, Enric

    2016-04-01

    One of the most important activities of the Regional Center for Northern Africa, Middle East and Europe of the World Meteorological Organization's Sand and Dust Storm Warning Advisory and Assessment System (WMO SDS-WAS, http://sds-was.aemet.es) is the dust model intercomparison and forecast evaluation, which is deemed an indispensable service to the users and an invaluable tool to assess model skills. Currently, the Regional Center collects daily dust forecasts from models run by nine partners (BSC, ECMWF, NASA, NCEP, SEEVCCC, EMA, CNR-ISAC, NOA and UK Met Office). A multi-model ensemble has also been set up in an effort to provide added-value products to the users. The first problem to address the dust model evaluation is the scarcity of suitable routine observations near the Sahara, the world's largest source of mineral dust. The present contribution presents preliminary results of dust model evaluation using new observational datasets. The current routine evaluation of dust predictions is focused on total-column dust optical depth (DOD) and uses remote-sensing retrievals from sun-photometric (AERONET) and satellite (MODIS) measurements. However, most users of dust forecasts are interested in the concentration near the surface (in the air we breathe) rather than in the total column content. Therefore, evaluation of the predicted surface concentration is also necessary. In this context, the initiative of the African Monsoon Interdisciplinary Analysis (AMMA) International Program to establish permanent measuring stations in the Sahel is extremely important. Tapered Element Oscillating Microbalance (TEOM) monitors continuously record PM10 in M'Bour (Senegal); Cinzana (Mali) and Banizoumbou (Niger). This surface model evaluation is complemented with the PM10 observation from the Air Quality Control and Monitoring Network (AQCMN) of the Canary Islands (Spain). The region, located in the sub-tropical Eastern Atlantic (roughly 100 km west of the Moroccan coast), is

  12. Intrinsic protein fluorescence interferes with detection of tear glycoproteins in SDS-polyacrylamide gels using extrinsic fluorescent dyes.

    PubMed

    Zhao, Zhenjun; Aliwarga, Yulina; Willcox, Mark D P

    2007-12-01

    Intrinsic protein fluorescence may interfere with the visualization of proteins after SDS-polyacrylamide electrophoresis. In an attempt to analyze tear glycoproteins in gels, we ran tear samples and stained the proteins with a glycoprotein-specific fluorescent dye. The fluorescence detected was not limited to glycoproteins. There was strong intrinsic fluorescence of proteins normally found in tears after soaking the gels in 40% methanol plus 1-10% acetic acid and, to a lesser extent, in methanol or acetic acid alone. Nanograms of proteins gave visible native fluorescence and interfere with extrinsic fluorescent dye detection. Poly-L-lysine, which does not contain intrinsically fluorescent amino acids, did not fluoresce.

  13. Intrinsic Protein Fluorescence Interferes with Detection of Tear Glycoproteins in SDS-Polyacrylamide Gels Using Extrinsic Fluorescent Dyes

    PubMed Central

    Zhao, Zhenjun; Aliwarga, Yulina; Willcox, Mark DP

    2007-01-01

    Intrinsic protein fluorescence may interfere with the visualization of proteins after SDS-polyacrylamide electrophoresis. In an attempt to analyze tear glycoproteins in gels, we ran tear samples and stained the proteins with a glycoprotein-specific fluorescent dye. The fluorescence detected was not limited to glycoproteins. There was strong intrinsic fluorescence of proteins normally found in tears after soaking the gels in 40% methanol plus 1–10% acetic acid and, to a lesser extent, in methanol or acetic acid alone. Nanograms of proteins gave visible native fluorescence and interfere with extrinsic fluorescent dye detection. Poly-L-lysine, which does not contain intrinsically fluorescent amino acids, did not fluoresce. PMID:18166676

  14. Effects of temperature and SDS on the structure of beta-glycosidase from the thermophilic archaeon Sulfolobus solfataricus.

    PubMed Central

    D'auria, S; Barone, R; Rossi, M; Nucci, R; Barone, G; Fessas, D; Bertoli, E; Tanfani, F

    1997-01-01

    The effects of temperature and SDS on the three-dimensional organization and secondary structure of beta-glycosidase from the thermophilic archaeon Sulfolobus solfataricus were investigated by CD, IR spectroscopy and differential scanning calorimetry. CD spectra in the near UV region showed that the detergent caused a remarkable change in the protein tertiary structure, and far-UV CD analysis revealed only a slight effect on secondary structure. Infrared spectroscopy showed that low concentrations of the detergent (up to 0.02%) induced slight changes in the enzyme secondary structure, whereas high concentrations caused the alpha-helix content to increase at high temperatures and prevented protein aggregation. PMID:9169619

  15. Web Pages for Your Classroom: The Easy Way!

    ERIC Educational Resources Information Center

    McCorkle, Sandra K.

    This book provides the classroom teacher or librarian with templates and instructions for creating Web pages for use with middle school or high school students. The pages can then be used for doing research projects or other types of projects that familiarize students with the power, flexibility, and usefulness of the Web. Part I, Technology in…

  16. Attention: Page has moved

    Science.gov Websites

    Information CPC Web Team USA.gov is the U.S. Government's official Web portal to all Federal, state and local government Web resources and services. This page has moved In about 10 seconds you will be transferred to its

  17. Purification and characterization of Bacillus cereus protease suitable for detergent industry.

    PubMed

    Prakash, Monika; Banik, Rathindra Mohan; Koch-Brandt, Claudia

    2005-12-01

    An extracellular alkaline protease from an alkalophilic bacterium, Bacillus cereus, was produced in a large amount by the method of extractive fermentation. The protease is thermostable, pH tolerant, and compatible with commercial laundry detergents. The protease purified and characterized in this study was found to be superior to endogenous protease already present in commercial laundry detergents. The enzyme was purified to homogeneity by ammonium sulfate precipitation, concentration by ultrafiltration, anion-exchange chromatography, and gel filtration. The purified enzyme had a specific activity of 3256.05 U/mg and was found to be a monomeric protein with a molecular mass of 28 and 31 kDa, as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and nondenaturing PAGE, respectively. Its maximum protease activity against casein was found to be at pH 10.5 and 50 degrees C. Proteolytic activity of the enzyme was detected by casein and gelatin zymography, which gave a very clear protease activity zone on gel that corresponded to the band obtained on SDS-PAGE and nondenaturing PAGE with a molecular mass of nearly 31 kDa. The purified enzyme was analyzed through matrix-assisted laser desorption ionization-time-of-flight-mass spectrometry (MALDI-TOF-MS) and identified as a subtilisin class of protease. Specific serine protease inhibitors, suggesting the presence of serine residues at the active site, inhibited the enzyme significantly.

  18. Starch granule-associated proteins of hull-less barley (Hordeum vulgare L.) from the Qinghai-Tibet Plateau in China.

    PubMed

    Wang, Chun-Ping; Pan, Zhi-Fen; Nima, Zha-Xi; Tang, Ya-Wei; Cai, Peng; Liang, Jun-Jun; Deng, Guang-Bing; Long, Hai; Yu, Mao-Qun

    2011-03-15

    The starch granule-associated proteins (SGAPs) are the minor components of the starch granules and a majority of them are believed to be starch biosynthetic enzymes. The Qinghai-Tibet Plateau in China, one of the centres of origin of cultivated barley, is abundant in hull-less barley resources which exhibit high polymorphism in SGAPs. The SGAPs of hull-less barley from Qinghai-Tibet Plateau were analysed by one-dimensional (1-D) SDS-PAGE, 2-D PAGE and ESI-Q-TOF MS/MS. In the 1-D SDS-PAGE gel, four proteins including a 80 kDa starch synthase, actin, actin 4 and ATP synthase β-subunit were identified as novel SGAPs. A total of six different bands were identified as starch granule-bound starch synthase I (GBSSI) and the segregation of the novel GBSSI bands in F(1) and F(2) seeds derived from yf127 × yf70 was in accordance with Mendel's law. In the 2-D PAGE gel, 92 spots were identified as 42 protein species which could be classified into 15 functional groups. Thirteen protein species were identified as SGAPs for the first time and multiple spots were identified as GBSSI. This study revealed novel SGAPs in hull-less barley from the Qinghai-Tibet Plateau in China and these will be significant in further studies of starch biosynthesis in barley. Copyright © 2011 Society of Chemical Industry.

  19. Navigation To and From a Page: Which Links Get Clicked From Where

    EPA Pesticide Factsheets

    Use Google Analytics navigation summary data to find out what page users most frequently click your Contact Us link from (Previous Page Path), or which links on your homepage are popular or unpopular (Next Page Path).

  20. Is Domain Highlighting Actually Helpful in Identifying Phishing Web Pages?

    PubMed

    Xiong, Aiping; Proctor, Robert W; Yang, Weining; Li, Ninghui

    2017-06-01

    To evaluate the effectiveness of domain highlighting in helping users identify whether Web pages are legitimate or spurious. As a component of the URL, a domain name can be overlooked. Consequently, browsers highlight the domain name to help users identify which Web site they are visiting. Nevertheless, few studies have assessed the effectiveness of domain highlighting, and the only formal study confounded highlighting with instructions to look at the address bar. We conducted two phishing detection experiments. Experiment 1 was run online: Participants judged the legitimacy of Web pages in two phases. In Phase 1, participants were to judge the legitimacy based on any information on the Web page, whereas in Phase 2, they were to focus on the address bar. Whether the domain was highlighted was also varied. Experiment 2 was conducted similarly but with participants in a laboratory setting, which allowed tracking of fixations. Participants differentiated the legitimate and fraudulent Web pages better than chance. There was some benefit of attending to the address bar, but domain highlighting did not provide effective protection against phishing attacks. Analysis of eye-gaze fixation measures was in agreement with the task performance, but heat-map results revealed that participants' visual attention was attracted by the highlighted domains. Failure to detect many fraudulent Web pages even when the domain was highlighted implies that users lacked knowledge of Web page security cues or how to use those cues. Potential applications include development of phishing prevention training incorporating domain highlighting with other methods to help users identify phishing Web pages.

  1. Caseoperoxidase, Mixed β-Casein-SDS-Hemin-Imidazole Complex: A Nano Artificial Enzyme

    PubMed Central

    Moosavi-Movahedi, Zainab; Gharibi, Hussein; Hadi-Alijanvand, Hamid; Akbarzadeh, Mohammad; Esmaili, Mansoore; Atri, Maliheh S.; Sefidbakht, Yahya; Bohlooli, Mousa; Nazari, Khodadad; Javadian, Soheila; Hong, Jun; Saboury, Ali A.; Sheibani, Nader; Moosavi-Movahedi, Ali A.

    2016-01-01

    A novel peroxidase-like artificial enzyme, named “caseoperoxidase”, was biomimetically designed using a nano artificial amino acid apo-protein hydrophobic pocket. This four-component nano artificial enzyme containing heme-imidazole-β-casein-SDS exhibited high activity growth and kcat performance towards the native horseradish peroxidase (HRP) demonstrated by the steady state kinetics using UV-Vis spectrophotometry. The hydrophobicity and secondary structure of the caseoperoxidase were studied by ANS fluorescence and circular dichroism spectroscopy. Camel β-casein (Cβ-casein), with a flexible structure and exalted hydrophobicity, was selected as an appropriate apo-protein for the heme active site using a homology modeling method. Heme docking into the newly obtained Cβ-casein structure indicated one heme was mainly incorporated with Cβ-casein. The presence of a main electrostatic site for the active site in the Cβ-casein was also confirmed by experimental methods through Wyman binding potential and isothermal titration calorimetry. The existence of Cβ-casein protein in this biocatalyst lowered the suicide inactivation, and indicated that the obtained structure has a good protective role for the heme active-site. Additional further experiments confirmed the retention of caseoperoxidase structure and function as an artificial enzyme. PMID:25562503

  2. Caseoperoxidase, mixed β-casein-SDS-hemin-imidazole complex: a nano artificial enzyme.

    PubMed

    Moosavi-Movahedi, Zainab; Gharibi, Hussein; Hadi-Alijanvand, Hamid; Akbarzadeh, Mohammad; Esmaili, Mansoore; Atri, Maliheh S; Sefidbakht, Yahya; Bohlooli, Mousa; Nazari, Khodadad; Javadian, Soheila; Hong, Jun; Saboury, Ali A; Sheibani, Nader; Moosavi-Movahedi, Ali A

    2015-01-01

    A novel peroxidase-like artificial enzyme, named "caseoperoxidase", was biomimetically designed using a nano artificial amino acid apo-protein hydrophobic pocket. This four-component nano artificial enzyme containing heme-imidazole-β-casein-SDS exhibited high activity growth and k(cat) performance toward the native horseradish peroxidase demonstrated by the steady state kinetics using UV-vis spectrophotometry. The hydrophobicity and secondary structure of the caseoperoxidase were studied by ANS fluorescence and circular dichroism spectroscopy. Camel β-casein (Cβ-casein) was selected as an appropriate apo-protein for the heme active site because of its innate flexibility and exalted hydrophobicity. This selection was confirmed by homology modeling method. Heme docking into the newly obtained Cβ-casein structure indicated one heme was mainly incorporated with Cβ-casein. The presence of a main electrostatic site for the active site in the Cβ-casein was also confirmed by experimental methods through Wyman binding potential and isothermal titration calorimetry. The existence of Cβ-casein protein in this biocatalyst lowered the suicide inactivation and provided a suitable protective role for the heme active-site. Additional experiments confirmed the retention of caseoperoxidase structure and function as an artificial enzyme.

  3. European user trial of paging by satellite

    NASA Technical Reports Server (NTRS)

    Fudge, R. E.; Fenton, C. J.

    1990-01-01

    British Telecom conceived the idea of adapting their existing paging service, together with the use of existing terrestrial pagers, to yield a one way data (i.e., paging) satellite service to mobiles. The user trial of paging by satellites was successful. It demonstrated that services could be provided over a wide geographical area to low priced terminals. Many lessons were learned in unexpected areas. These include the need for extensive liaison with all users involved, especially the drivers, to ensure they understood the potential benefits. There was a significant desire for a return acknowledgement channel or even a return data channel. Above all there is a need to ensure that the equipment can be taken across European borders and legitimately used in all European countries. The next step in a marketing assessment would be to consider the impact of two way data messaging such as INMARSAT-C.

  4. Improving Interdisciplinary Provider Communication Through a Unified Paging System.

    PubMed

    Heidemann, Lauren; Petrilli, Christopher; Gupta, Ashwin; Campbell, Ian; Thompson, Maureen; Cinti, Sandro; Stewart, David A

    2016-06-01

    Interdisciplinary communication at a Veterans Affairs (VA) academic teaching hospital is largely dependent on alphanumeric paging, which has limitations as a result of one-way communication and lack of reliable physician identification. Adverse patient outcomes related to difficulty contacting the correct consulting provider in a timely manner have been reported. House officers were surveyed on the level of satisfaction with the current VA communication system and the rate of perceived adverse patient outcomes caused by potential delays within this system. Respondents were then asked to identify the ideal paging system. These results were used to develop and deploy a new Web site. A postimplementation survey was repeated 1 year later. This study was conducted as a quality improvement project. House officer satisfaction with the preintervention system was 3%. The majority used more than four modalities to identify consultants, with 59% stating that word of mouth was a typical source. The preferred mode of paging was the university hospital paging system, a Web-based program that is used at the partnering academic institution. Following integration of VA consulting services within the university hospital paging system, the level of satisfaction improved to 87%. Significant decreases were seen in perceived adverse patient outcomes (from 16% to 2%), delays in patient care (from 90% to 16%), and extended hospitalizations (from 46% to 4%). Our study demonstrates significant improvement in physician satisfaction with a newly implemented paging system that was associated with a decreased perceived number of adverse patient events and delays in care.

  5. Some Features of "Alt" Texts Associated with Images in Web Pages

    ERIC Educational Resources Information Center

    Craven, Timothy C.

    2006-01-01

    Introduction: This paper extends a series on summaries of Web objects, in this case, the alt attribute of image files. Method: Data were logged from 1894 pages from Yahoo!'s random page service and 4703 pages from the Google directory; an img tag was extracted randomly from each where present; its alt attribute, if any, was recorded; and the…

  6. Quality of drug information on the World Wide Web and strategies to improve pages with poor information quality. An intervention study on pages about sildenafil.

    PubMed

    Martin-Facklam, Meret; Kostrzewa, Michael; Martin, Peter; Haefeli, Walter E

    2004-01-01

    The generally poor quality of health information on the world wide web (WWW) has caused preventable adverse outcomes. Quality management of information on the internet is therefore critical given its widespread use. In order to develop strategies for the safe use of drugs, we scored general and content quality of pages about sildenafil and performed an intervention to improve their quality. The internet was searched with Yahoo and AltaVista for pages about sildenafil and 303 pages were included. For assessment of content quality a score based on accuracy and completeness of essential drug information was assigned. For assessment of general quality, four criteria were evaluated and their association with high content quality was determined by multivariate logistic regression analysis. The pages were randomly allocated to either control or intervention group. Evaluation took place before, as well as 7 and 22 weeks after an intervention which consisted of two letters with individualized feedback information on the respective page which were sent electronically to the address mentioned on the page. Providing references to scientific publications or prescribing information was significantly associated with high content quality (odds ratio: 8.2, 95% CI 3.2, 20.5). The intervention had no influence on general or content quality. To prevent adverse outcomes caused by misinformation on the WWW individualized feedback to the address mentioned on the page was ineffective. It is currently probably the most straight-forward approach to inform lay persons about indicators of high information quality, i.e. the provision of references.

  7. Quality of drug information on the World Wide Web and strategies to improve pages with poor information quality. An intervention study on pages about sildenafil

    PubMed Central

    Martin-Facklam, Meret; Kostrzewa, Michael; Martin, Peter; Haefeli, Walter E

    2004-01-01

    Aims The generally poor quality of health information on the world wide web (WWW) has caused preventable adverse outcomes. Quality management of information on the internet is therefore critical given its widespread use. In order to develop strategies for the safe use of drugs, we scored general and content quality of pages about sildenafil and performed an intervention to improve their quality. Methods The internet was searched with Yahoo and AltaVista for pages about sildenafil and 303 pages were included. For assessment of content quality a score based on accuracy and completeness of essential drug information was assigned. For assessment of general quality, four criteria were evaluated and their association with high content quality was determined by multivariate logistic regression analysis. The pages were randomly allocated to either control or intervention group. Evaluation took place before, as well as 7 and 22 weeks after an intervention which consisted of two letters with individualized feedback information on the respective page which were sent electronically to the address mentioned on the page. Results Providing references to scientific publications or prescribing information was significantly associated with high content quality (odds ratio: 8.2, 95% CI 3.2, 20.5). The intervention had no influence on general or content quality. Conclusions To prevent adverse outcomes caused by misinformation on the WWW individualized feedback to the address mentioned on the page was ineffective. It is currently probably the most straight-forward approach to inform lay persons about indicators of high information quality, i.e. the provision of references. PMID:14678344

  8. Book Holder And Page Turner For The Elderly And Handicapped

    NASA Technical Reports Server (NTRS)

    Kerley, James; Eklund, Wayne

    1993-01-01

    Device holds reading matter and facilitates page turning for person not having use of arms and hands. Accommodates variety of publication formats, whether book, magazine, or newspaper. Holder sits on hospital-bed table and adjusted to convenient viewing angle. Includes flat upright back support for reading matter, hinged base, and main bracket with bent-wire page holders. Top support on back extended for such large items as newspapers. Wings on back support extended for oversize materials. Reader turns page by gripping special rod via mouthpiece, applying friction cup at its tip to page, and manipulating rod. Mouthpiece wide and tapered so user grips with teeth and uses jaws to move it, rather than using tongue or lips. Helpful to older people, whose facial and mouth muscles weak.

  9. Network and User-Perceived Performance of Web Page Retrievals

    NASA Technical Reports Server (NTRS)

    Kruse, Hans; Allman, Mark; Mallasch, Paul

    1998-01-01

    The development of the HTTP protocol has been driven by the need to improve the network performance of the protocol by allowing the efficient retrieval of multiple parts of a web page without the need for multiple simultaneous TCP connections between a client and a server. We suggest that the retrieval of multiple page elements sequentially over a single TCP connection may result in a degradation of the perceived performance experienced by the user. We attempt to quantify this perceived degradation through the use of a model which combines a web retrieval simulation and an analytical model of TCP operation. Starting with the current HTTP/l.1 specification, we first suggest a client@side heuristic to improve the perceived transfer performance. We show that the perceived speed of the page retrieval can be increased without sacrificing data transfer efficiency. We then propose a new client/server extension to the HTTP/l.1 protocol to allow for the interleaving of page element retrievals. We finally address the issue of the display of advertisements on web pages, and in particular suggest a number of mechanisms which can make efficient use of IP multicast to send advertisements to a number of clients within the same network.

  10. La preparacion en ciencia de los candidatos a maestros del nivel elemental primario segun la reforma de la educacion cientifica en Puerto Rico: Una propuesta de secuencia curricular

    NASA Astrophysics Data System (ADS)

    Rodriguez Plaza, Evelyn

    El proposito de esta investigacion fue identificar los componentes de la preparacion en ciencia que deben recibir los estudiantes del Bachillerato en Artes en Educacion Elemental, Nivel Primario, de acuerdo a los documentos que dirigen la reforma de la educacion cientifica en Puerto Rico. Tambien, se identificaron los componentes de los cursos que forman parte de la preparacion en ciencia de estos estudiantes. Se compararon los componentes de la preparacion en ciencia y los componentes de los cursos para determinar congruencias y discrepancias. Con los datos recopilados se identificaron los componentes de los cursos de una secuencia curricular para la preparacion en ciencia de los candidatos a maestros del nivel elemental primario. La secuencia curricular que se propone en esta investigacion incluye cursos de contenido cientifico y de metodologia en la ensenanza de la ciencia disenados para satisfacer las necesidades de los candidatos. Se recomienda que en los procesos para el diseno, la implantacion y la evaluacion de estos cursos participen profesores de ciencia, profesores de educacion y maestros del nivel elemental primario. Todos los cursos de la secuencia curricular deben tener un enfoque constructivista. Las experiencias educativas que se incluyan en los cursos deben aspirar a desarrollar en los candidatos los atributos de la cultura cientifica y actitudes positivas hacia la ciencia y hacia la ensenanza de esta disciplina. El modelaje por parte de los profesores que ensenen los cursos de la secuencia curricular es fundamental en el desarrollo profesional de los candidatos. Se recomienda que en los cursos de contenido cientifico se estudien los conceptos y los conocimientos cientificos que forman parte del curriculo de Kindergarten a tercer grado de forma integrada y con una profundidad universitaria. Estos cursos deben tener un enfoque interdisciplinario e incluir el estudio de la naturaleza de la ciencia y un componente de laboratorio para desarrollar los

  11. [An evaluation of the quality of health web pages using a validated questionnaire].

    PubMed

    Conesa Fuentes, Maria del Carmen; Aguinaga Ontoso, Enrique; Hernández Morante, Juan José

    2011-01-01

    The objective of the present study was to evaluate the quality of general health information in Spanish language web pages, and the official Regional Services web pages from the different Autonomous Regions. It is a cross-sectional study. We have used a previously validated questionnaire to study the present state of the health information on Internet for a lay-user point of view. By mean of PageRank (Google®), we obtained a group of webs, including a total of 65 health web pages. We applied some exclusion criteria, and finally obtained a total of 36 webs. We also analyzed the official web pages from the different Health Services in Spain (19 webs), making a total of 54 health web pages. In the light of our data, we observed that, the quality of the general information health web pages was generally rather low, especially regarding the information quality. Not one page reached the maximum score (19 points). The mean score of the web pages was of 9.8±2.8. In conclusion, to avoid the problems arising from the lack of quality, health professionals should design advertising campaigns and other media to teach the lay-user how to evaluate the information quality. Copyright © 2009 Elsevier España, S.L. All rights reserved.

  12. An Analysis of Academic Library Web Pages for Faculty

    ERIC Educational Resources Information Center

    Gardner, Susan J.; Juricek, John Eric; Xu, F. Grace

    2008-01-01

    Web sites are increasingly used by academic libraries to promote key services and collections to teaching faculty. This study analyzes the content, location, language, and technological features of fifty-four academic library Web pages designed especially for faculty to expose patterns in the development of these pages.

  13. Washington VAAC Redirect Page

    Science.gov Websites

    National Environmental Satellite, Data, and Information Service Home Page Default Office of Satellite and Atlantic Composites Pacific Composites Satellite Services Argos DCS EMWIN GEONETCast Americas GOES DCS LRIT NOAA DRO Conference NOAASIS SARSAT ---- Satellite Information ---- GOES -- Satellite Status -- Special

  14. Proteomic Profiling of Rat Thyroarytenoid Muscle

    ERIC Educational Resources Information Center

    Welham, Nathan V.; Marriott, Gerard; Bless, Diane M.

    2006-01-01

    Purpose: Proteomic methodologies offer promise in elucidating the systemwide cellular and molecular processes that characterize normal and diseased thyroarytenoid (TA) muscle. This study examined methodological issues central to the application of 2-dimensional sodium dodecyl sulfate polyacrylamide gel electrophoresis (2D SDS-PAGE) to the study of…

  15. EF-2DE Analysis and Protein Identification

    USDA-ARS?s Scientific Manuscript database

    Isoelectric focusing followed by SDS-PAGE (IEF-2DE) separates proteins in a two-dimensional matrix of protein pI (Protein Isoelectric Point) and molecular weight (MW). The technique is particularly useful to distinguish protein isoforms (Radwan et al., 2012) and proteins that contain post-translatio...

  16. Regenerative Medicine and Restoration of Joint Function

    DTIC Science & Technology

    2013-10-01

    examined if the inability of adult human chondrocytes to form cartilage tissue is dependent on age or species differences between human and cows ...SDS-PAGE, and transferred to a nitrocellulose membrane by iblot (Invitrogen, US). Membranes were blocked in 1% milk for 1 h and incubated with

  17. A novel procedure for the extraction of protein deposits from soft hydrophilic contact lenses for analysis.

    PubMed

    Keith, D; Hong, B; Christensen, M

    1997-05-01

    A quick, simple, and efficient extraction technique was developed for the removal of protein from soft hydrophilic contact lenses. An extraction solvent consisting of a 50:50 mix of 0.2% trifluoroacetic acid and acetonitrile was used to remove protein from in vitro laboratory-deposited and human-worn contact lenses. The protein removed was analyzed using HPLC, bicinchoninic acid (BCA) analysis, and SDS-PAGE gel electreophoresis. Extraction efficiency for lysozyme from laboratory-deposited Group IV lenses was determined to be approximately 100%. Group IV human-worn contact lenses were extracted and analyzed for lysozyme by HPLC and total protein by bicinchoninic acid (BCA) analysis. Groups I, II, III, and IV contact lenses deposited with an artificial tear protein solution and human-worn lenses were extracted and analyzed by SDS-PAGE gel electreophoresis and micro-BCA. The ACN/TFA procedure offers a simple, quick, and efficient extraction technique for removal of protein from contact lenses for subsequent analysis.

  18. Effects of industrial cashew nut processing on anacardic acid content and allergen recognition by IgE.

    PubMed

    Mattison, Christopher P; Malveira Cavalcante, Jéfferson; Izabel Gallão, Maria; Sousa de Brito, Edy

    2018-02-01

    Cashew nuts are important both nutritionally and industrially, but can also cause food allergies in some individuals. The present study aimed to assess the effect(s) of industrial processing on anacardic acids and allergens present in cashew nuts. Sample analyses were performed using liquid chromatography coupled with mass spectrometry, SDS-PAGE and immunoassay. The anacardic acid concentration ranged from 6.2 to 82.6mg/g during processing, and this variation was attributed to cashew nut shell liquid incorporation during storage and humidification. Dehydrated and selected samples did not significantly differ in anacardic acid content, having values similar to the raw sample. SDS-PAGE and immunoassay analysis with rabbit polyclonal sera and human IgE indicated only minor differences in protein solubility and antibody binding following processing steps. The findings indicate that appreciable amounts of anacardic acid remain in processed nuts, and that changes to cashew allergens during industrial processing may only mildly affect antibody recognition. Published by Elsevier Ltd.

  19. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Yip, Wingkip; Dong, Jianguo,; Yang, Shang Fa

    Tomato ACC synthase is inactivated by its substrate SAM, with the moiety of aminobutyrate being covalently linked to ACC synthase during the catalytic reactions. A partial purified ACC synthase (the catalytic activity 100 {mu}mol/h{center dot}mg protein) from pellets of apple extract was incubated with (3,4{sup 14}C) SAM. Only one radioactive peak was revealed in a C-4 reverse phase HPLC and one radioactive band on SDS-PAGE with an M.W. of 48 kDa. Apple ACC synthase in native form is resistant to V8, {alpha}-chromtrypsin and carboxylpeptidase A digestion, but effectively inactivated by trypsin and ficin, as demonstrated by both the activity assaymore » and SAM labeling. The radioactive protein cut from the SDS-PAGE was injected to three mice, two of the mice showed responses to the protein in western blot analysis. The antibodies from mice is currently under characterization.« less

  20. Children monosensitized to pine nuts have similar patterns of sensitization.

    PubMed

    Novembre, Elio; Mori, Francesca; Barni, Simona; Ferrante, Giuliana; Pucci, Neri; Ballabio, Cinzia; Uberti, Francesca; Penas, Elena; Restani, Patrizia

    2012-12-01

    Several cases of pine nut allergies and anaphylaxis have been reported in the literature, but only few pine nut allergens have been characterized. The aim of this research is to identify through immunoelectrophoretic techniques the major pine nut allergens in a group of children monosensitized to pine nuts. We studied five children with pine nut allergies and no other sensitization to food except to pine nuts, confirmed by in vivo (prick test, prick-to-prick) and in vitro tests (specific IgE determinations [CAP-FEIA]). The protein profile of pine nuts was analyzed by Sodium Dodecyl sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE). Immunoblotting was performed after incubation of membranes with the sera from the children included in the present study. Immunoblotting (SDS-PAGE) demonstrated five similar bands between 6 and 47 kDa in all the subjects studied. These bands should be considered the potential allergens for pine nut allergic children. © 2012 John Wiley & Sons A/S.

  1. Effects of heat and high-pressure treatments on the solubility and immunoreactivity of almond proteins.

    PubMed

    Zhang, Yan; Zhang, Jieqiong; Sheng, Wei; Wang, Shuo; Fu, Tong-Jen

    2016-05-15

    The effects of dry and moist heat, autoclave sterilization and high-pressure treatment on the biochemical characteristics and immunological properties of almond proteins were investigated. Changes in the solubility and immunoreactivity of almond proteins extracted from treated almond flour were evaluated using a total protein assay, indirect competitive inhibition enzyme-linked immunosorbent assay (IC-ELISA), and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Almond proteins were stable during dry-heat treatment at temperatures below 250°C. Dry heat at 400°C, boiling, autoclave sterilization and high-pressure treatment in the presence of water at ⩾ 500 MPa greatly reduced the solubility and immunoreactivity of almond proteins. SDS-PAGE revealed that the protein profiles of almond flour samples treated under these conditions also changed significantly. The synergistic effects of heat, pressure and the presence of water contributed to significant changes in solubility and immunoreactivity of almond proteins. Copyright © 2015 Elsevier Ltd. All rights reserved.

  2. Electrophoresis characterisation of protein as a method to establish the entomological origin of stingless bee honeys.

    PubMed

    Ramón-Sierra, Jesús Manuel; Ruiz-Ruiz, Jorge Carlos; de la Luz Ortiz-Vázquez, Elizabeth

    2015-09-15

    Increasing production of stingless-bee honey and the prospect of broader marker for natural and organic products indicate the need to establish parameters to determinate the entomological origin and authenticity of honey. In this research, honeys of Apis mellifera, Melipona beecheii and Trigona spp. were collected in Yucatan, Mexico. Stingless-bee honeys contained more water and less total sugars and reducing sugars. SDS-PAGE patterns show distinctive bands for each kind of honey. The SDS-PAGE pattern of A. mellifera proteins honey showed three bands with molecular weights between 10.2 and 74.8kDa, there were five proteins bands in M. beecheii honey with molecular weights between 6.1 and 97.0kDa and nine for Trigona spp. proteins between 9.3 and 86.7kDa. Conventional physicochemical parameters along with electrophoresis profiles of stingless-bee honeys proteins could be an alternative for determination of entomological origin. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Pulsed electric field (PEF)-induced aggregation between lysozyme, ovalbumin and ovotransferrin in multi-protein system.

    PubMed

    Wu, Li; Zhao, Wei; Yang, Ruijin; Yan, Wenxu

    2015-05-15

    The aggregation of multi-proteins is of great interest in food processing and a good understanding of the formation of aggregates during PEF processing is needed for the application of the process to pasteurize protein-based foods. The aggregates formation of a multi-protein system (containing ovalbumin, ovotransferrin and lysozyme) was studied through turbidity, size exclusion chromatography and SDS-PAGE patterns for interaction studies and binding forces. Results from size exclusion chromatography indicated that there was no soluble aggregates formed during PEF processing. The existence of lysozyme was important to form insoluble aggregates in the chosen ovalbumin solution. The results of SDS-PAGE patterns indicated that lysozyme was prone to precipitate, and was relatively the higher component of aggregates. Citric acid could be effective in inhibiting lysozyme from interacting with other proteins during PEF processing. Blocking the free sulphydryl by N-ethylmaleimide (NEM) did not affect aggregation inhibition. Copyright © 2014 Elsevier Ltd. All rights reserved.

  4. Absence of sugars in electrophoretically purified cytochrome b5 demonstrated by combined gas chromatography-mass spectrometry

    PubMed Central

    1981-01-01

    The problem of determining small but significant amounts of carbohydrates, in purified proteins, has been studied using the membrane protein, cytochrome b5. A newly developed method that involves direct gas chromatography-mass spectrometry of sugars obtained by hydrolysis of proteins purified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) allows the identification and determination of small amounts of carbohydrates (e.g., 20 micrograms of glycoprotein containing a minimum of 0.1% monosaccharide), even in the presence of relatively high amounts of impurities. Application of this method to cytochrome b5 fragments obtained by tryptic digestion from rat liver microsomes and purified by combined gel filtration and ion exchange chromatography, followed by SDS PAGE, has consistently yielded values below 0.07 mol of the individual sugars and aminosugars per mole cytochrome b5. It is concluded that cytochrome b5, at least its trypsin-released major amino- terminal fragment, is not constitutively glycosylated. PMID:7251667

  5. Marination and Physicochemical Characteristics of Vacuum-aged Duck Breast Meat.

    PubMed

    Khan, Muhammad Issa; Lee, Hyun Jung; Kim, Hyun-Joo; Young, Hae In; Lee, Haelim; Jo, Cheorun

    2016-11-01

    We investigated marinade absorption and physicochemical characteristics of vacuum-aged duck breasts that were halved and individually vacuum-packed for chiller aging at 4°C for 14 d. One half was marinated for 0, 7, or 14 d, while the second half was used as a control. Marinade absorption, cooking loss, cooking yield, texture profile, pH, color, protein solubility, and thiobarbituric acid reactive substances (TBARS) values were evaluated, and protein sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was performed. Marinade absorption and pH did not vary significantly after 14 d of aging. Marination increased the pH, color (a* and b*) values, and cooking yield and reduced cooking loss. TBARS values significantly increased with aging time, but were significantly reduced by marination. Myofibril and total protein solubility increased with aging and marination, while SDS-PAGE showed protein degradation. Hence, aging and marination can be used simultaneously to improve physicochemical quality and cooking yield of vacuum-aged duck breast.

  6. Purification and properties of fructosyl lysine oxidase from Fusarium oxysporum S-1F4.

    PubMed

    Sakai, Y; Yoshida, N; Isogai, A; Tani, Y; Kato, N

    1995-03-01

    Fructosyl lysine oxidase (FLOD) was examined for its use in the enzymatic measurement of the level of glycated albumin in blood serum. To isolate microorganisms having such an enzyme activity, we used N epsilon-fructosyl N alpha-Z-lysine (epsilon-FL) as a sole nitrogen source in the enrichment culture medium. The isolated fungus, strain S-1F4, showed a high FLOD activity in the cell-free extract and was identified as Fusarium oxysporum. FLOD was purified to an apparent homogeneity on SDS-PAGE. The molecular mass of the subunit was 50 kDa on SDS-PAGE and seemed to exist in a monomeric form. The enzyme had an absorption spectrum characteristic of a flavoprotein and the flavin was found to be covalently bound to the enzyme. The enzyme acted against N epsilon-fructosyl N alpha-Z-lysine and N alpha-fructosyl N epsilon-Z-lysine and showed specificity for fructosyl lysine residues.

  7. Characterization of viral proteins of Oryctes baculovirus and comparison between two geographical isolates.

    PubMed

    Mohan, K S; Gopinathan, K P

    1989-01-01

    Bacilliform Oryctes baculovirus particles have been visualized in electron micrographs of midgut sections from virus infected Oryctes rhinoceros beetles. Morphologically the Indian isolate (Oryctes baculovirus, KI) resembled the previously reported Oryctes baculovirus, isolate PV505. The constituent proteins of baculovirus KI have been analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and by Western blots using polyclonal antibodies raised against the complete viral particles, as probes. A total of forty eight viral proteins have been identified. Fourteen viral proteins were located on the viral envelope. Among the proteins constituting the nucleocapsid, three were located internally within the capsid. A 23.5 kDa protein was tightly associated with viral DNA in the nucleocapsid core. Two envelope and seven capsid proteins of KI and PV505 revealed differences in SDS-PAGE profiles and glycosylation patterns. Immunoblotting of KI and PV505 proteins with anti KI antiserum demonstrated antigenic differences between the two viral isolates.

  8. Functionality of ovalbumin during Chinese steamed bread-making processing.

    PubMed

    Sang, Shangyuan; Zhang, Huang; Xu, Lei; Chen, Yisheng; Xu, Xueming; Jin, Zhengyu; Yang, Na; Wu, Fengfeng; Li, Dandan

    2018-07-01

    Hen egg is commonly used in some cereal-based food, including cakes and bread. Ovalbumin, one of the major components of egg white protein, can affect the performance of the food product. The interaction between ovalbumin and gluten protein and its effect on property of dough and quality of Chinese steamed bread was investigated in this study. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) patterns indicated that ovalbumin was surprisingly not incorporated in glutenins by covalent bond, whereas size-exclusion high-performance liquid chromatography showed that glutenin macropolymer content in glutenins increased slightly. Furthermore, dynamic rheology experiments indicated ovalbumin led to a decrease inG' andG″ of dough. Based on molecular dynamic simulation and SDS-PAGE results, it was inferred that ovalbumin was not hydrolyzed by endopeptidases during dough fermentation and crosslinked to gluten proteins during steaming. Finally, ovalbumin improved maximum dough height (Hm) during dough development and specific volume of Chinese steamed bread. Copyright © 2018 Elsevier Ltd. All rights reserved.

  9. Development of a wheat-Aegilops searsii substitution line with positively affecting Chinese steamed bread quality.

    PubMed

    Du, Xuye; Ma, Xin; Min, Jingzhi; Zhang, Xiaocun; Jia, Zhenzhen

    2018-03-01

    A wheat- Aegilops searsii substitution line GL1402, in which chromosome 1B was substituted with 1S s from Ae. searsii , was developed and detected using SDS-PAGE and GISH. The SDS-PAGE analysis showed that the HMW-GS encoded by the Glu-B1 loci of Chinese Spring was replaced by the HMW-GS encoded by the Glu-1S s loci of Ae. searsii . Glutenin macropolymer (GMP) investigation showed that GL1402 had a much higher GMP content than Chinese Spring did. A dough quality comparison of GL1402 and Chinese Spring indicated that GL1402 showed a significantly higher protein content and middle peak time (MPT), and a smaller right peak slope (RPS). Quality tests of Chinese steamed bread (CSB) showed that the GL1402 also produced good steamed bread quality. These results suggested that the substitution line is a valuable breeding material for improving the wheat processing quality.

  10. Antioxidant and ACE-inhibitory activities of hemp (Cannabis sativa L.) protein hydrolysates produced by the proteases AFP, HT, Pro-G, actinidin and zingibain.

    PubMed

    Teh, Sue-Siang; Bekhit, Alaa El-Din A; Carne, Alan; Birch, John

    2016-07-15

    Hemp protein isolates (HPIs) were hydrolysed by proteases (AFP, HT, ProG, actinidin and zingibain). The enzymatic hydrolysis of HPIs was evaluated through the degree of hydrolysis and SDS-PAGE profiles. The bioactive properties of the resultant hydrolysates (HPHs) were accessed through ORAC, DPPḢ scavenging and ACE-inhibitory activities. The physical properties of the resultant HPHs were evaluated for their particle sizes, zeta potential and surface hydrophobicity. HT had the highest rate of caseinolytic activity at the lowest concentration (0.1 mg mL(-1)) compared to other proteases that required concentration of 100 mg mL(-1) to achieve their maximum rate of caseinolytic activity. This led to the highest degree of hydrolysis of HPIs by HT in the SDS-PAGE profiles. Among all proteases and substrates, HT resulted in the highest bioactivities (ORAC, DPPḢ scavenging and ACE-inhibitory activities) generated from alkali extracted HPI in the shortest time (2 h) compared to the other protease preparations. Copyright © 2016 Elsevier Ltd. All rights reserved.

  11. Cardiology Patient Page: Electronic Cigarettes

    MedlinePlus

    ... of the American Heart Association Cardiology Patient Page Electronic Cigarettes Rachel A. Grana , Pamela M. Ling , Neal ... Footnotes References Figures & Tables Info & Metrics eLetters Introduction Electronic cigarettes (E-cigarettes) are devices that deliver nicotine ...

  12. Soluble scute proteins of healthy and ill desert tortoises (Gopherus agassizii)

    USGS Publications Warehouse

    Homer, B.L.; Li, C.; Berry, K.H.; Denslow, N.D.; Jacobson, E.R.; Sawyer, R.H.; Williams, J.E.

    2001-01-01

    Objectives - To characterize protein composition of shell scute of desert tortoises and to determine whether detectable differences could be used to identify healthy tortoises from tortoises with certain illnesses. Animals - 20 desert tortoises. Procedures - Complete postmortem examinations were performed on all tortoises. Plastron scute proteins were solubilized, scute proteins were separated by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and proteins were analyzed, using densitometry. Two-dimensional immobilized pH gradient-PAGE (2D IPG-PAGE) and immunoblot analysis, using polyclonal antisera to chicken-feather ?? keratin and to alligator-scale ?? keratin, were conducted on representative samples. The 14-kd proteins were analyzed for amino acid composition. Results - The SDS-PAGE and densitometry revealed 7 distinct bands, each with a mean relative protein concentration of > 1 %, ranging from 8 to 47 kd, and a major protein component of approximately 14 kd that constituted up to 75% of the scute protein. The 2D IPG-PAGE revealed additional distinct 62-and 68-kd protein bands. On immunoblot analysis, the 14-, 32-, and 45-kd proteins reacted with both antisera. The 14-kd proteins had an amino acid composition similar to that of chicken ?? keratins. There was a substantial difference in the percentage of the major 14-kd proteins from scute of ill tortoises with normal appearing shells, compared with 14-kd proteins of healthy tortoises. Conclusions and Clinical Relevance - The major protein components of shell scute of desert tortoises have amino acid composition and antigenic features of ?? keratins. Scute protein composition may be altered in tortoises with certain systemic illnesses.

  13. 46. Photograph of a published page. 'OPERATIONS IN INCORPORATION BUILDINGS: ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    46. Photograph of a published page. 'OPERATIONS IN INCORPORATION BUILDINGS: HOLSTON DEFENSE CORPORATION. 'HOLSTON ARMY AMMUNITION PLANT.' Page 17. (no date). - Holston Army Ammunition Plant, RDX-and-Composition-B Manufacturing Line 9, Kingsport, Sullivan County, TN

  14. Practice Audit in Gastroenterology (PAGE) program: A novel approach to continuing professional development

    PubMed Central

    Armstrong, David; Hollingworth, Roger; Gardiner, Tara; Klassen, Michael; Smith, Wendy; Hunt, Richard H; Barkun, Alan; Gould, Michael; Leddin, Desmond

    2006-01-01

    BACKGROUND: Practice audit is an important component of continuing professional development that may more readily be undertaken if it were less complex. This qualitative study assessed the use of personal digital assistants to facilitate data collection and review. METHODS: Personal digital assistants programmed with standard questionnaires related to upper gastrointestinal endoscopies (Practice Audit in Gastroenterology-Endoscopy [‘PAGE-Endo’]) and colonoscopies (PAGE-Colonoscopy [‘PAGE-Colo’]) were provided to Canadian gastroenterologists, surgeons and internists. Over a three-week audit period, participants recorded indications, and the expected (E) and reported (R) findings for each procedure. Thereafter, participants recorded compliance with reporting, the ease of use and value of the PAGE program, and their willingness to perform another audit. RESULTS: Over 15 to 18 months, 173 participants completed PAGE-Endo (6168 procedures) and 111 completed PAGE-Colo (4776 procedures). Most respondents noted that PAGE was easy to use (99%), beneficial (88% to 95%), and that they were willing undertake another audit (92% to 95%). In PAGE-Endo, alarm features were prevalent (55%), but major reported findings were less common than expected: esophagitis (E 29.9%, R 14.8%), esophageal stricture (E 8.3%, R 3.6%), gastric ulcer (E 17.0%, R 4.7%), gastric cancer (E 4.3%, R 1.0%) and duodenal ulcer (E 11.5%, R 5.7%). In PAGE-Colo, more colonoscopies were performed for symptom investigation (55%) than for screening (25%) or surveillance (20%). There were marked interprovincial variations with respect to sedation, biopsies and technical aspects of colonoscopy. CONCLUSION: Secure, real-time data entry with review of aggregate and individual data in the PAGE program provided an acceptable, straightforward methodology for accredited practice audit activities. PAGE has considerable potential for continuing professional development in gastroenterology and other specialties

  15. Accelerating Demand Paging for Local and Remote Out-of-Core Visualization

    NASA Technical Reports Server (NTRS)

    Ellsworth, David

    2001-01-01

    This paper describes a new algorithm that improves the performance of application-controlled demand paging for the out-of-core visualization of data sets that are on either local disks or disks on remote servers. The performance improvements come from better overlapping the computation with the page reading process, and by performing multiple page reads in parallel. The new algorithm can be applied to many different visualization algorithms since application-controlled demand paging is not specific to any visualization algorithm. The paper includes measurements that show that the new multi-threaded paging algorithm decreases the time needed to compute visualizations by one third when using one processor and reading data from local disk. The time needed when using one processor and reading data from remote disk decreased by up to 60%. Visualization runs using data from remote disk ran about as fast as ones using data from local disk because the remote runs were able to make use of the remote server's high performance disk array.

  16. 45. Photograph of a published page. OPERATIONS IN 'H' OR ...

    Library of Congress Historic Buildings Survey, Historic Engineering Record, Historic Landscapes Survey

    45. Photograph of a published page. OPERATIONS IN 'H' OR DEWATERING BUILDING: HOLSTON DEFENSE CORPORATION. 'HOLSTON ARMY AMMUNITION PLANT.' Page 16. (no date). - Holston Army Ammunition Plant, RDX-and-Composition-B Manufacturing Line 9, Kingsport, Sullivan County, TN

  17. Effect of Pulsed Ultraviolet Light and High Hydrostatic Pressure on the Antigenicity of Almond Protein Extracts.

    USDA-ARS?s Scientific Manuscript database

    The efficacy of pulsed ultraviolet light (PUV) and high hydrostatic pressure (HHP) on reducing the IgE binding to the almond extracts, was studied using SDS-PAGE, Western Blot, and ELISA probed with human plasma containing IgE antibodies to almond allergens, and a polyclonal antibody against almond ...

  18. Guaiacol Peroxidase Zymography for the Undergraduate Laboratory

    ERIC Educational Resources Information Center

    Wilkesman, Jeff; Castro, Diana; Contreras, Lellys M.; Kurz, Liliana

    2014-01-01

    This laboratory exercise presents a novel way to introduce undergraduate students to the specific detection of enzymatic activity by electrophoresis. First, students prepare a crude peroxidase extract and then analyze the homogenate via electrophoresis. Zymography, that is, a SDS-PAGE method to detect enzyme activity, is used to specifically…

  19. Effects of salinity, pH and temperature on the re-establishment of bioluminescence and copper or SDS toxicity in the marine dinoflagellate Pyrocystis lunula using bioluminescence as an endpoint

    USGS Publications Warehouse

    Craig, J.M.; Klerks, P.L.; Heimann, K.; Waits, J.L.

    2003-01-01

    Pyrocystis lunula is a unicellular, marine, photoautotrophic, bioluminescent dinoflagellate. This organism is used in the Lumitox ?? bioassay with inhibition of bioluminescence re-establishment as the endpoint. Experiments determined if acute changes in pH, salinity, or temperature had an effect on the organisms' ability to re-establish bioluminescence, or on the bioassay's potential to detect sodium dodecyl sulfate (SDS) and copper toxicity. The re-establishment of bioluminescence itself was not very sensitive to changes in pH within the pH 6-10 range, though reducing pH from 8 to levels below 6 decreased this capacity. Increasing the pH had little effect on Cu or SDS toxicity, but decreasing the pH below 7 virtually eliminated the toxicity of either compound in the bioassay. Lowering the salinity from 33 to 27??? or less resulted in a substantial decrease in re-establishment of bioluminescence, while increasing the salinity to 43 or 48 ??? resulted in a small decline. Salinity had little influence on the bioassay's quantification of Cu toxicity, while the data showed a weak negative relationship between SDS toxicity and salinity. Re-establishment of bioluminescence showed a direct dependence on temperature, but only at 10??C did temperature have an obvious effect on the toxicity of Cu in this bioassay. ?? 2003 Elsevier Science Ltd. All rights reserved.

  20. Sources and Coverage of Medical News on Front Pages of US Newspapers

    PubMed Central

    Lai, William Y. Y.; Lane, Trevor; Jones, Alison

    2009-01-01

    Background Medical news that appears on newspaper front pages is intended to reach a wide audience, but how this type of medical news is prepared and distributed has not been systematically researched. We thus quantified the level of visibility achieved by front-page medical stories in the United States and analyzed their news sources. Methodology Using the online resource Newseum, we investigated front-page newspaper coverage of four prominent medical stories, and a high-profile non-medical news story as a control, reported in the US in 2007. Two characteristics were quantified by two raters: which newspaper titles carried each target front-page story (interrater agreement, >96%; kappa, >0.92) and the news sources of each target story (interrater agreement, >94%; kappa, >0.91). National rankings of the top 200 US newspapers by audited circulation were used to quantify the extent of coverage as the proportion of the total circulation of ranked newspapers in Newseum. Findings In total, 1630 front pages were searched. Each medical story appeared on the front pages of 85 to 117 (67.5%–78.7%) ranked newspaper titles that had a cumulative daily circulation of 23.1 to 33.4 million, or 61.8% to 88.4% of all newspapers. In contrast, the non-medical story achieved front-page coverage in 152 (99.3%) newspaper titles with a total circulation of 41.0 million, or 99.8% of all newspapers. Front-page medical stories varied in their sources, but the Washington Post, Los Angeles Times, New York Times and the Associated Press together supplied 61.7% of the total coverage of target front-page medical stories. Conclusion Front-page coverage of medical news from different sources is more accurately revealed by analysis of circulation counts rather than of newspaper titles. Journals wishing to widen knowledge of research news and organizations with important health announcements should target at least the four dominant media organizations identified in this study. PMID:19724643

  1. Market study: Tactile paging system

    NASA Technical Reports Server (NTRS)

    1977-01-01

    A market survey was conducted regarding the commercialization potential and key market factors relevant to a tactile paging system for deaf-blind people. The purpose of the tactile paging system is to communicate to the deaf-blind people in an institutional environment. The system consists of a main console and individual satellite wrist units. The console emits three signals by telemetry to the wrist com (receiving unit) which will measure approximately 2 x 4 x 3/4 inches and will be fastened to the wrist by a strap. The three vibration signals are fire alarm, time period indication, and a third signal which will alert the wearer of the wrist com to the fact that the pin on the top of the wrist is emitting a morse coded message. The Morse code message can be felt and recognized with the finger.

  2. JavaScript: Convenient Interactivity for the Class Web Page.

    ERIC Educational Resources Information Center

    Gray, Patricia

    This paper shows how JavaScript can be used within HTML pages to add interactive review sessions and quizzes incorporating graphics and sound files. JavaScript has the advantage of providing basic interactive functions without the use of separate software applications and players. Because it can be part of a standard HTML page, it is…

  3. Attention: Page has moved

    Science.gov Websites

    Weather Service NWS logo - Click to go to the NWS home page Climate Prediction Center Site Map News with the new address. NOAA/ National Weather Service National Centers for Environmental Prediction Climate Prediction Center 5200 Auth Road Camp Springs, Maryland 20746 Climate Prediction Center Web Team

  4. Allergy to grass pollen: mapping of Dactylis glomerata and Phleum pratense allergens for dogs by two-dimensional immunoblotting

    PubMed Central

    Marques, Andreia Grilo; Pereira, Luísa Maria Dotti Silva; Semião-Santos, Saul José; Bento, Ofélia Pereira

    2017-01-01

    Introduction Much less is known about grass-pollen allergens to dogs, when compared with humans. Genetic-based patterns might play an important role in sensitization profiles, conditioning the success of allergen-specific immunotherapy. Aim Mapping of Dactylis glomerata (D. glomerata) and Phleum pratense (P. pratense) allergens for grass pollen-sensitized atopic dogs, for better understanding how individual allergograms may influence the response to grass-pollen immunotherapy. Material and methods To identify D. glomerata and P. pratense allergoms for dogs, 15 individuals allergic to grass pollen and sensitized to D. glomerata and P. pratense were selected. D. glomerata and P. pratense proteomes were separated by isoelectric focusing (IEF), one-dimensional (1-D) and two-dimensional (2-D) sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Separated proteins were blotted onto Polyvinylidene difluoride (PVDF) membranes and allergens were identified by patient sera IgE in Western Blotting (WB). Results In D. glomerata, 17 allergens were identified from IEF and 11 from 1-D SDS-PAGE, while from P. pratense, 18 and 6 allergens were identified, respectively. From 2-D SDS-PAGE 13 spots were identified from D. glomerata and 27 from P. pratense. Conclusions Several similarities were found between dog and human D. glomerata and P. pratense sensitization profiles but no relationship between clinical signs and a specific pattern of allergen recognition was observed. Similarities were found in each patient pattern of sensitization between D. glomerata and P. pratense, also suggesting cross-reactive phenomena. Further molecular epidemiology approach is needed to understand the role of the sensitization pattern in allergen-specific immunotherapy effectiveness in grass-pollen allergic dogs. PMID:28261033

  5. Deep proteomic profiling of vasopressin-sensitive collecting duct cells. I. Virtual Western blots and molecular weight distributions.

    PubMed

    Yang, Chin-Rang; Tongyoo, Pumipat; Emamian, Milad; Sandoval, Pablo C; Raghuram, Viswanathan; Knepper, Mark A

    2015-12-15

    The mouse mpkCCD cell line is a continuous cultured epithelial cell line with characteristics of renal collecting duct principal cells. This line is widely used to study epithelial transport and its regulation. To provide a data resource useful for experimental design and interpretation in studies using mpkCCD cells, we have carried out "deep" proteomic profiling of these cells using three levels of fractionation (differential centrifugation, SDS-PAGE, and HPLC) followed by tandem mass spectrometry to identify and quantify proteins. The analysis of all resulting samples generated 34.6 gigabytes of spectral data. As a result, we identified 6,766 proteins in mpkCCD cells at a high level of stringency. These proteins are expressed over eight orders of magnitude of protein abundance. The data are provided to users as a public data base (https://helixweb.nih.gov/ESBL/Database/mpkFractions/). The mass spectrometry data were mapped back to their gel slices to generate "virtual Western blots" for each protein. For most of the 6,766 proteins, the apparent molecular weight from SDS-PAGE agreed closely with the calculated molecular weight. However, a substantial fraction (>15%) of proteins was found to run aberrantly, with much higher or much lower mobilities than predicted. These proteins were analyzed to identify mechanisms responsible for altered mobility on SDS-PAGE, including high or low isoelectric point, high or low hydrophobicity, physiological cleavage, residence in the lysosome, posttranslational modifications, and expression of alternative isoforms due to alternative exon usage. Additionally, this analysis identified a previously unrecognized isoform of aquaporin-2 with apparent molecular mass <20 kDa. Copyright © 2015 the American Physiological Society.

  6. Deep proteomic profiling of vasopressin-sensitive collecting duct cells. I. Virtual Western blots and molecular weight distributions

    PubMed Central

    Yang, Chin-Rang; Tongyoo, Pumipat; Emamian, Milad; Sandoval, Pablo C.; Raghuram, Viswanathan

    2015-01-01

    The mouse mpkCCD cell line is a continuous cultured epithelial cell line with characteristics of renal collecting duct principal cells. This line is widely used to study epithelial transport and its regulation. To provide a data resource useful for experimental design and interpretation in studies using mpkCCD cells, we have carried out “deep” proteomic profiling of these cells using three levels of fractionation (differential centrifugation, SDS-PAGE, and HPLC) followed by tandem mass spectrometry to identify and quantify proteins. The analysis of all resulting samples generated 34.6 gigabytes of spectral data. As a result, we identified 6,766 proteins in mpkCCD cells at a high level of stringency. These proteins are expressed over eight orders of magnitude of protein abundance. The data are provided to users as a public data base (https://helixweb.nih.gov/ESBL/Database/mpkFractions/). The mass spectrometry data were mapped back to their gel slices to generate “virtual Western blots” for each protein. For most of the 6,766 proteins, the apparent molecular weight from SDS-PAGE agreed closely with the calculated molecular weight. However, a substantial fraction (>15%) of proteins was found to run aberrantly, with much higher or much lower mobilities than predicted. These proteins were analyzed to identify mechanisms responsible for altered mobility on SDS-PAGE, including high or low isoelectric point, high or low hydrophobicity, physiological cleavage, residence in the lysosome, posttranslational modifications, and expression of alternative isoforms due to alternative exon usage. Additionally, this analysis identified a previously unrecognized isoform of aquaporin-2 with apparent molecular mass <20 kDa. PMID:26310816

  7. Development and characterization of an allergoid of cat dander for immunotherapy.

    PubMed

    Sola, J P; Pedreño, Y; Cerezo, A; Peñalver-Mellado, M

    2018-01-13

    Allergy to cats is a frequent cause of sensitization to indoor allergens and currently there are few alternatives to specific immunotherapy with cat native extracts. The objective is to develop and characterize a new allergoid to increase the tools available for use in clinical practice. The allergoid cat dander extract (ACD) was developed from a native cat dander extract (NCD) by modification with glutaraldehyde, and the optimal process control was determined by SDS-PAGE, DOT BLOT and determination of free amine groups. The ACD was characterized in protein profile by SDS-PAGE, size exclusion chromatography (SEC) and peptide footprint. The allergenic profile of ACD was determined by immunoblot, IgE CAP inhibition and IgG competition ELISA. The major allergen content in NCD was obtained by the ELISA sandwich protocol and was extrapolated to ACD. The control process determined the optimal development of the allergoid. The ACD obtained contains 182.28μg/mg of protein and 11.90μg/mg of Fel d 1. SDS-PAGE and SEC confirmed the presence of high molecular weight proteins in ACD, and the peptide footprint showed the presence of Fel d 1 and Fel d 7. The high degree of polymerization was evidenced with the determination of the reduction of lysine residues in the allergoid, resulting 91.96%. The ACD showed a significant loss of allergenicity respect to NCD, while the IgG-binding capacity was maintained. The ACD obtained presents a good safety profile, so would be a good alternative for treatment of cat allergy. Copyright © 2017 SEICAP. Published by Elsevier España, S.L.U. All rights reserved.

  8. The envelopes of amphibian oocytes: physiological modifications in Bufo arenarum.

    PubMed

    Barisone, Gustavo A; Albertali, Isabel E; Sánchez, Mercedes; Cabada, Marcelo O

    2003-02-11

    A characterization of the Amphibian Bufo arenarum oocyte envelope is presented. It was made in different functional conditions of the oocyte: 1) when it has been released into the coelomic cavity during ovulation (surrounded by the coelomic envelope, (CE), 2) after it has passed through the oviduct and is deposed (surrounded by the viteline envelope, (VE), and 3) after oocyte activation (surrounded by the fertilization envelope, (FE). The characterization was made by SDS-PAGE followed by staining for protein and glycoproteins. Labeled lectins were used to identify glycosidic residues both in separated components on nitrocellulose membranes or in intact oocytes and embryos. Proteolytic properties of the content of the cortical granules were also analyzed. After SDS-PAGE of CE and VE, a different protein pattern was observed. This is probably due to the activity of a protease present in the pars recta of the oviduct. Comparison of the SDS-PAGE pattern of VE and FE showed a different mobility for one of the glycoproteins, gp75. VE and FE proved to have different sugar residues in their oligosaccharide chains. Mannose residues are only present in gp120 of the three envelopes. N-acetyl-galactosamine residues are present in all of the components, except for gp69 in the FE. Galactose residues are present mainly in gp120 of FE. Lectin-binding assays indicate the presence of glucosamine, galactose and N-acetyl galactosamine residues and the absence (or non-availability) of N-acetyl-glucosamine or fucose residues on the envelopes surface. The cortical granule product (CGP) shows proteolytic activity on gp75 of the VE.

  9. The envelopes of amphibian oocytes: physiological modifications in Bufo arenarum

    PubMed Central

    Barisone, Gustavo A; Albertali, Isabel E; Sánchez, Mercedes; Cabada, Marcelo O

    2003-01-01

    A characterization of the Amphibian Bufo arenarum oocyte envelope is presented. It was made in different functional conditions of the oocyte: 1) when it has been released into the coelomic cavity during ovulation (surrounded by the coelomic envelope, (CE), 2) after it has passed through the oviduct and is deposed (surrounded by the viteline envelope, (VE), and 3) after oocyte activation (surrounded by the fertilization envelope, (FE). The characterization was made by SDS-PAGE followed by staining for protein and glycoproteins. Labeled lectins were used to identify glycosidic residues both in separated components on nitrocellulose membranes or in intact oocytes and embryos. Proteolytic properties of the content of the cortical granules were also analyzed. After SDS-PAGE of CE and VE, a different protein pattern was observed. This is probably due to the activity of a protease present in the pars recta of the oviduct. Comparison of the SDS-PAGE pattern of VE and FE showed a different mobility for one of the glycoproteins, gp75. VE and FE proved to have different sugar residues in their oligosaccharide chains. Mannose residues are only present in gp120 of the three envelopes. N-acetyl-galactosamine residues are present in all of the components, except for gp69 in the FE. Galactose residues are present mainly in gp120 of FE. Lectin-binding assays indicate the presence of glucosamine, galactose and N-acetyl galactosamine residues and the absence (or non-availability) of N-acetyl-glucosamine or fucose residues on the envelopes surface. The cortical granule product (CGP) shows proteolytic activity on gp75 of the VE. PMID:12694627

  10. Analysis of Soluble Proteins in Natural Cordyceps sinensis from Different Producing Areas by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis and Two-dimensional Electrophoresis.

    PubMed

    Li, Chun-Hong; Zuo, Hua-Li; Zhang, Qian; Wang, Feng-Qin; Hu, Yuan-Jia; Qian, Zheng-Ming; Li, Wen-Jia; Xia, Zhi-Ning; Yang, Feng-Qing

    2017-01-01

    As one of the bioactive components in Cordyceps sinensis (CS), proteins were rarely used as index components to study the correlation between the protein components and producing areas of natural CS. Protein components of 26 natural CS samples produced in Qinghai, Tibet, and Sichuan provinces were analyzed and compared to investigate the relationship among 26 different producing areas. Proteins from 26 different producing areas were extracted by Tris-HCl buffer with Triton X-100, and separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE). The SDS-PAGE results indicated that the number of protein bands and optical density curves of proteins in 26 CS samples was a bit different. However, the 2-DE results showed that the numbers and abundance of protein spots in protein profiles of 26 samples were obviously different and showed certain association with producing areas. Based on the expression values of matched protein spots, 26 batches of CS samples can be divided into two main categories (Tibet and Qinghai) by hierarchical cluster analysis. The number of protein bands and optical density curves of proteins in 26 Cordyceps sinensis samples were a bit different on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profilesNumbers and abundance of protein spots in protein profiles of 26 samples were obvious different on two-dimensional electrophoresis mapsTwenty-six different producing areas of natural Cordyceps sinensis samples were divided into two main categories (Tibet and Qinghai) by Hierarchical cluster analysis based on the values of matched protein spots. Abbreviations Used : SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis, 2-DE: Two-dimensional electrophoresis, Cordyceps sinensis : CS, TCMs: Traditional Chinese medicines.

  11. Code AI Personal Web Pages

    NASA Technical Reports Server (NTRS)

    Garcia, Joseph A.; Smith, Charles A. (Technical Monitor)

    1998-01-01

    The document consists of a publicly available web site (george.arc.nasa.gov) for Joseph A. Garcia's personal web pages in the AI division. Only general information will be posted and no technical material. All the information is unclassified.

  12. Reporting on post-menopausal hormone therapy: an analysis of gynaecologists' web pages.

    PubMed

    Bucksch, Jens; Kolip, Petra; Deitermann, Bernhilde

    2004-01-01

    The present study was designed to analyse Web pages of German gynaecologists with regard to postmenopausal hormone therapy (HT). There is a growing body of evidence, that the overall health risks of HT exceed the benefits. Making one's own informed choice has become a central concern for menopausal women. The Internet is an important source of health information, but the quality is often dubious. The study focused on the analysis of basic criteria such as last modification date and quality of the HT information content. The results of the Women's Health Initiative Study (WHI) were used as a benchmark. We searched for relevant Web pages by entering a combination of key words (9 x 13 = 117) into the search engine www.google.de. Each Web page was analysed using a standardized questionnaire. The basic criteria and the quality of content on each Web page were separately categorized by two evaluators. Disagreements were resolved by discussion. Of the 97 websites identified, basic criteria were not met by the majority. For example, the modification date was displayed by only 23 (23.7%) Web pages. The quality of content of most Web pages regarding HT was inaccurate and incomplete. Whilst only nine (9.3%) took up a balanced position, 66 (68%) recommended HT without any restrictions. In 22 cases the recommendation was indistinct and none of the sites refused HT. With regard to basic criteria, there was no difference between HT-recommending Web pages and sites with balanced position. Evidence-based information resulting from the WHI trial was insufficiently represented on gynaecologists' Web pages. Because of the growing number of consumers looking online for health information, the danger of obtaining harmful information has to be minimized. Web pages of gynaecologists do not appear to be recommendable for women because they do not provide recent evidence-based findings about HT.

  13. Partial purification and characterization of polyphenoloxidase from culinary-medicinal Royal Sun mushroom (the Himematsutake), Agaricus brasiliensis S. Wasser et al. (Agaricomycetideae).

    PubMed

    Matsumoto-Akanuma, Akiko; Akanuma, Satoshi; Motoi, Masuro; Yamagishi, Akihiko; Ohno, Naohito

    2011-01-01

    The Royal Sun mushroom, the Himematsutake culinary-medicinal mushroom, Agaricus brasiliensis has several polyphenoloxidase activities in a broad sense. Here we report the partial purification of tyrosinase-type polyphenoloxidase (PPO). PPO is purified from A. brasiliensis without browning using a two-phase partitioning with Triton X-114 and ammonium sulfate fractionation. Partially denaturing SDS-PAGE (sodium dodecyl sulfate-polyacrylamide electrophoresis) staining with L-3,4-dihydroxyphenylalanine was performed and the indicated molecular sizes were approximately 70 kDa and 45 kDa. The purified enzyme is in its latent state and can be activated maximally in the presence of 1.6 mM sodium dodecyl sulfate (SDS). This enzyme catalyzes two distinct reactions, monophenolase and diphenolase activity, and the monophenolase activity showed a lag time typical of polyphenoloxidase. The K(m) value for 4-tert-butylcatechol was quite similar in the presence and absence of SDS, but the apparent V(max) value was increased 2.0-fold by SDS. Mimosine was a typical competitive inhibitor with K(i) values of 138.2 microM and 281.0 microM n the presence and absence of SDS, respectively.

  14. Creating a Facebook Page for the Seismological Society of America

    NASA Astrophysics Data System (ADS)

    Newman, S. B.

    2009-12-01

    In August, 2009 I created a Facebook “fan” page for the Seismological Society of America. We had been exploring cost-effective options for providing forums for two-way communication for some months. We knew that a number of larger technical societies had invested significant sums of money to create customized social networking sites but that a small society would need to use existing low-cost software options. The first thing I discovered when I began to set up the fan page was that an unofficial SSA Facebook group already existed, established by Steven J. Gibbons, a member in Norway. Steven had done an excellent job of posting material about SSA. Partly because of the existing group, the official SSA fan page gained fans rapidly. We began by posting information about our own activities and then added links to activities in the broader geoscience community. While much of this material also appeared on our website and in our publication, Seismological Research Letters (SRL), the tone on the FB page is different. It is less formal with more emphasis on photos and links to other sites, including our own. Fans who are active on FB see the posts as part of their social network and do not need to take the initiative to go to the SSA site. Although the goal was to provide a forum for two-way communication, our initial experience was that people were clearly reading the page but not contributing content. This appears to be case with fan pages of sister geoscience societies. FB offers some demographic information to fan site administrators. In an initial review of the demographics it appeared that fans were younger than the overall demographics of the Society. It appeared that a few of the fans are not members or even scientists. Open questions are: what content will be most useful to fans? How will the existence of the page benefit the membership as a whole? Will the page ultimately encourage two-way communication as hoped? Web 2.0 is generating a series of new

  15. The impact of visual layout factors on performance in Web pages: a cross-language study.

    PubMed

    Parush, Avi; Shwarts, Yonit; Shtub, Avy; Chandra, M Jeya

    2005-01-01

    Visual layout has a strong impact on performance and is a critical factor in the design of graphical user interfaces (GUIs) and Web pages. Many design guidelines employed in Web page design were inherited from human performance literature and GUI design studies and practices. However, few studies have investigated the more specific patterns of performance with Web pages that may reflect some differences between Web page and GUI design. We investigated interactions among four visual layout factors in Web page design (quantity of links, alignment, grouping indications, and density) in two experiments: one with pages in Hebrew, entailing right-to-left reading, and the other with English pages, entailing left-to-right reading. Some performance patterns (measured by search times and eye movements) were similar between languages. Performance was particularly poor in pages with many links and variable densities, but it improved with the presence of uniform density. Alignment was not shown to be a performance-enhancing factor. The findings are discussed in terms of the similarities and differences in the impact of layout factors between GUIs and Web pages. Actual or potential applications of this research include specific guidelines for Web page design.

  16. Effects of Fe deficiency on the protein profile of Brassica napus phloem sap

    USDA-ARS?s Scientific Manuscript database

    The aim of this work was to study the effect of Fe deficiency on the protein profile of phloem sap exudates from Brassica napus using 2-DE (IEF-SDS PAGE). The experiment was repeated thrice and two technical replicates per treatment were done. Two hundred sixty-three spots were consistently detected...

  17. Metallothionein quantification in clams by reversed-phase high-performance liquid chromatography coupled to fluorescence detection after monobromobimane derivatization.

    PubMed

    Alhama, José; Romero-Ruiz, Antonio; López-Barea, Juan

    2006-02-24

    In this paper, we describe a highly specific, sensitive and reliable method for total metallothionein (MT) quantification by RP-HPLC coupled to fluorescence detection following reaction with monobromobimane of thiols from metal-depleted MT after heat-denaturation of extracts in the presence of sodium dodecyl sulphate (SDS). SDS-polyacrylamide gel electrophoresis (SDS-PAGE) confirmed the identity of the peak resolved (t(R)=16.44) with MT: a highly fluorescent protein of approximately 8.3 kDa, in agreement with the high thiol content and low MT size. Other heat-resistant and Cys-containing proteins of 35 kDa were efficiently separated. The new method was successfully used to quantify MT content in digestive gland of clams from southern Spanish coastal sites with different metal levels, and is proposed as a tool for using MTs as biomarker in monitoring programmes.

  18. Portable exhauster POR-007/Skid E and POR-008/Skid F storage plan

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Nelson, O.D.

    1998-07-25

    This document provides storage requirements for 1,000 CFM portable exhausters POR-O07/Skid E and POR-008/Skid F. These requirements are presented in three parts: preparation for storage, storage maintenance and testing, and retrieval from storage. The exhauster component identification numbers listed in this document contain the prefix POR-007 or POR-008 depending on which exhauster is being used.

  19. SSE Data and Information Page

    Atmospheric Science Data Center

    2018-04-04

    Surface meteorology and Solar Energy (SSE) Data and Information A new POWER home page ... The Release 6.0 Surface meteorology and Solar Energy (SSE) data set contains parameters formulated for assessing and designing renewable energy systems. This latest release contains new parameters based on ...

  20. NOAA History - Main Page

    Science.gov Websites

    NOAA History Banner gold bar divider home - takes you to index page about the site contacts noaa americas science and service noaa legacy 1807 - 2007 NOAA History is an intrinsic part of the history of Initiative scroll divider More NOAA History from Around the Nation scroll divider drawing of a tornado NOAA

  1. Analysis of cytoskeletal proteins in posterior capsule opacification after implantation of acrylic and hydrogel intraocular lenses.

    PubMed

    Matsushima, Hiroyuki; Mukai, Kouichiro; Obara, Yoshitaka; Yoshida, Shinichiro; Clark, John I

    2004-01-01

    To analyze selected lens cytoskeletal proteins in posterior capsule opacification (PCO) 2 weeks after intraocular lens (IOL) implantation in rabbits. Department of Ophthalmology, Dokkyo University School of Medicine, Tochigi, Japan. Eight 10-week-old albino rabbits were prepared and anesthetized for phacoemulsification and aspiration of the crystalline lens and implantation of an acrylic or a hydrogel IOL. Two weeks postoperatively, the rabbits were killed and the IOLs removed for immunohistochemistry. Deparaffinized tissue sections were processed with antibodies against alpha-smooth muscle actin (alpha-SMA) and beta-crystallin to observe the types of PCO with the 2 IOL types. The proteins in the PCO tissue and the normal lens were homogenized, centrifuged, and analyzed using SDS-polyacrylamide gel electrophoresis (SDS-PAGE) densitometric analysis and Western immunoblotting for actin and vimentin. Immunohistochemistry demonstrated a fibroblastic cell type expressing alpha-SMA and partial regeneration of epithelial cells, resulting in a lenticular structure that stained irregularly for beta-crystallin. The immunoreactivity of fibroblast-like cells to beta-crystallin appeared weaker than that of the regenerated lenticular structure. SDS-PAGE showed variability in the content of cytoskeletal proteins in the insoluble fractions of the PCO. Degradation of the cytoskeletal components was greater with the acrylic IOL than with the hydrogel IOL. Cytoskeletal proteins expressed during the formation of PCO and IOL implantation may have potential as therapeutic target proteins to improve the biocompatibility of IOLs.

  2. The structure and dynamics of Nano Particles encapsulated by the SDS monolayer collapse at the water/TCE interface

    NASA Astrophysics Data System (ADS)

    Shi, Wenxiong

    2016-11-01

    The super-saturated surfactant monolayer collapses with the nanoparticles (NPs) at the water/trichloroethylene (TCE) interface are investigated using molecular dynamics (MD) simulations. The results show that sodium alkyl sulfate (SDS) monolayer collapse is initiated by buckling and followed primarily by budding and the bud encapsulating the NPs and oil molecules. The developed bud detaches from the monolayer into a water phase and forms the swollen micelle emulsion with NPs and oil molecules. We investigate the wavelength of the initial budding and the theoretical description of the budding process. The wavelength of the monolayer increases with bending modulus. The energy barrier of the budding can be easily overcome by thermal fluctuation energy, which indicates that budding process proceeds rapidly.

  3. Simple and rapid method for isolation and quantitation of polyhydroxyalkanoate by SDS-sonication treatment.

    PubMed

    Arikawa, Hisashi; Sato, Shunsuke; Fujiki, Tetsuya; Matsumoto, Keiji

    2017-08-01

    We developed a new method for isolation and quantitation of polyhydroxyalkanoate (PHA) from culture broth. In this method, the cells were sonicated in sodium dodecyl sulfate (SDS) solution and centrifuged to recover PHA. The recovered PHA was rinsed with deionized water and ethanol, and then weighed after drying. Hazardous chemicals such as chloroform, methanol, and sulfuric acid were not used, and no expensive analytical instruments were needed. We applied this method to Cupriavidus necator culture broths that included various amounts of poly(3-hydroxybutyrate) (PHB) or poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx) from flasks and jar fermentors. The quantitation by this method was practical for use with a wide range of production amounts and PHA monomer compositions compared to the conventional whole-cell methanolysis method with gas chromatographic analysis, and besides, the recovered PHAs were adequately pure (≥96% purity). Therefore, this new method would be valuable not only for quantitation of PHA but also for preparation of samples to characterize their mechanical properties. Copyright © 2017 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.

  4. A 7 ke-SD-FWC 1.2 e-RMS Temporal Random Noise 128×256 Time-Resolved CMOS Image Sensor With Two In-Pixel SDs for Biomedical Applications.

    PubMed

    Seo, Min-Woong; Kawahito, Shoji

    2017-12-01

    A large full well capacity (FWC) for wide signal detection range and low temporal random noise for high sensitivity lock-in pixel CMOS image sensor (CIS) embedded with two in-pixel storage diodes (SDs) has been developed and presented in this paper. For fast charge transfer from photodiode to SDs, a lateral electric field charge modulator (LEFM) is used for the developed lock-in pixel. As a result, the time-resolved CIS achieves a very large SD-FWC of approximately 7ke-, low temporal random noise of 1.2e-rms at 20 fps with true correlated double sampling operation and fast intrinsic response less than 500 ps at 635 nm. The proposed imager has an effective pixel array of and a pixel size of . The sensor chip is fabricated by Dongbu HiTek 1P4M 0.11 CIS process.

  5. Home Page: The Mode of Transport through the Information Superhighway

    NASA Technical Reports Server (NTRS)

    Lujan, Michelle R.

    1995-01-01

    The purpose of the project with the Aeroacoustics Branch was to create and submit a home page for the internet about branch information. In order to do this, one must also become familiar with the way that the internet operates. Learning HyperText Markup Language (HTML), and the ability to create a document using this language was the final objective in order to place a home page on the internet (World Wide Web). A manual of instructions regarding maintenance of the home page, and how to keep it up to date was also necessary in order to provide branch members with the opportunity to make any pertinent changes.

  6. 14 CFR 389.26 - Special rules for tariff page filings.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 14 Aeronautics and Space 4 2011-01-01 2011-01-01 false Special rules for tariff page filings. 389.26 Section 389.26 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION... § 389.26 Special rules for tariff page filings. (a) Tariffs issued by carriers. The filing fee for...

  7. 14 CFR 389.26 - Special rules for tariff page filings.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 14 Aeronautics and Space 4 2010-01-01 2010-01-01 false Special rules for tariff page filings. 389.26 Section 389.26 Aeronautics and Space OFFICE OF THE SECRETARY, DEPARTMENT OF TRANSPORTATION... § 389.26 Special rules for tariff page filings. (a) Tariffs issued by carriers. The filing fee for...

  8. Evaluating Information Quality: Hidden Biases on the Children's Web Pages

    ERIC Educational Resources Information Center

    Kurubacak, Gulsun

    2006-01-01

    As global digital communication continues to flourish, the Children's Web pages become more critical for children to realize not only the surface but also breadth and deeper meanings in presenting these milieus. These pages not only are very diverse and complex but also enable intense communication across social, cultural and political…

  9. Confirmation of the genome position and mass of the viral protien, VP3, of Solenopsis Invicta Virus 1 (Picornavirales: Dicistroviridae) infecting the Red Imported Fire Ant (Hymenoptera: Formicidae)

    USDA-ARS?s Scientific Manuscript database

    A predicted peptide sequence corresponding to the capsid protein VP3 of SINV-1 was used to prepare a polyclonal antibody preparation and probe SDS-PAGE separated proteins from SINV-1 particles and S. invicta ants. The SINV-1 VP3 antibody preparation recognized a 24 kDa protein from denatured SINV-1...

  10. Psychometric properties of the AIR Self-Determination Scale: the Chinese version (AIR SDS-C) for Chinese people with intellectual disabilities.

    PubMed

    Wong, P K S; Wong, D F K; Zhuang, X Y; Liu, Y

    2017-03-01

    The construct of self-determination has received considerable attention in the international field of intellectual disabilities (ID). Recently, there has been a rapid development of this construct in Chinese societies including Hong Kong. However, there is no locally validated instrument to measure self-determination in people with ID. This article explains the validation process of the AIR Self-Determination Scale - Chinese version (AIR SDS-C) adapted from the 24-item AIR Self-Determination Scale, developed by Wolman and his colleagues, which is used in school setting. People with mild/moderate ID aged 15 years or above were recruited from special schools and social services units in different regions of Hong Kong. Factor analysis and reliability tests were conducted. Data for a total of 356 participants were used for the analysis. A confirmatory factor analysis was performed to test the factorial construct, and Mplus 7.0 was used for the analysis. The factor structure proposed in the original English version was supported by the data, and all factor loadings were between 0.42 and 0.76. The whole scale achieved good reliability (Cronbach's α = 0.88 and ω = 0.90). The AIR SDS-C appears to be a valid and reliable scale. This study examined adult groups as well as student groups. The application of the scale can thus be extended to a wider population. The implications for theory building and practice are discussed. © 2016 MENCAP and International Association of the Scientific Study of Intellectual and Developmental Disabilities and John Wiley & Sons Ltd.

  11. Characterizing Microseismicity at the Newberry Volcano Geothermal Site using PageRank

    NASA Astrophysics Data System (ADS)

    Aguiar, A. C.; Myers, S. C.

    2015-12-01

    The Newberry Volcano, within the Deschutes National Forest in Oregon, has been designated as a candidate site for the Department of Energy's Frontier Observatory for Research in Geothermal Energy (FORGE) program. This site was stimulated using high-pressure fluid injection during the fall of 2012, which generated several hundred microseismic events. Exploring the spatial and temporal development of microseismicity is key to understanding how subsurface stimulation modifies stress, fractures rock, and increases permeability. We analyze Newberry seismicity using both surface and borehole seismometers from the AltaRock and LLNL seismic networks. For our analysis we adapt PageRank, Google's initial search algorithm, to evaluate microseismicity during the 2012 stimulation. PageRank is a measure of connectivity, where higher ranking represents highly connected windows. In seismic applications connectivity is measured by the cross correlation of 2 time windows recorded on a common seismic station and channel. Aguiar and Beroza (2014) used PageRank based on cross correlation to detect low-frequency earthquakes, which are highly repetitive but difficult to detect. We expand on this application by using PageRank to define signal-correlation topology for micro-earthquakes, including the identification of signals that are connected to the largest number of other signals. We then use this information to create signal families and compare PageRank families to the spatial and temporal proximity of associated earthquakes. Studying signal PageRank will potentially allow us to efficiently group earthquakes with similar physical characteristics, such as focal mechanisms and stress drop. Our ultimate goal is to determine whether changes in the state of stress and/or changes in the generation of subsurface fracture networks can be detected using PageRank topology. This work performed under the auspices of the U.S. Department of Energy by Lawrence Livermore National Laboratory under

  12. Best Practices for Searchable Collection Pages

    EPA Pesticide Factsheets

    Searchable Collection pages are stand-alone documents that do not have any web area navigation. They should not recreate existing content on other sites and should be tagged with quality metadata and taxonomy terms.

  13. World Wide Web home page for the South Platte NAWQA

    USGS Publications Warehouse

    Qi, Sharon L.; Dennehy, Kevin F.

    1997-01-01

    A World Wide Web home page for the U.S. Geological Survey's (USGS) National Water-Quality Assessment (NAWQA) Program, South Platte River Basin study is now online. The home page includes information about the basinwide investigation and provides viewing and downloading access to physical, chemical, and biological data collected by the study team.

  14. An efficient scheme for automatic web pages categorization using the support vector machine

    NASA Astrophysics Data System (ADS)

    Bhalla, Vinod Kumar; Kumar, Neeraj

    2016-07-01

    In the past few years, with an evolution of the Internet and related technologies, the number of the Internet users grows exponentially. These users demand access to relevant web pages from the Internet within fraction of seconds. To achieve this goal, there is a requirement of an efficient categorization of web page contents. Manual categorization of these billions of web pages to achieve high accuracy is a challenging task. Most of the existing techniques reported in the literature are semi-automatic. Using these techniques, higher level of accuracy cannot be achieved. To achieve these goals, this paper proposes an automatic web pages categorization into the domain category. The proposed scheme is based on the identification of specific and relevant features of the web pages. In the proposed scheme, first extraction and evaluation of features are done followed by filtering the feature set for categorization of domain web pages. A feature extraction tool based on the HTML document object model of the web page is developed in the proposed scheme. Feature extraction and weight assignment are based on the collection of domain-specific keyword list developed by considering various domain pages. Moreover, the keyword list is reduced on the basis of ids of keywords in keyword list. Also, stemming of keywords and tag text is done to achieve a higher accuracy. An extensive feature set is generated to develop a robust classification technique. The proposed scheme was evaluated using a machine learning method in combination with feature extraction and statistical analysis using support vector machine kernel as the classification tool. The results obtained confirm the effectiveness of the proposed scheme in terms of its accuracy in different categories of web pages.

  15. Content and Design Features of Academic Health Sciences Libraries' Home Pages.

    PubMed

    McConnaughy, Rozalynd P; Wilson, Steven P

    2018-01-01

    The goal of this content analysis was to identify commonly used content and design features of academic health sciences library home pages. After developing a checklist, data were collected from 135 academic health sciences library home pages. The core components of these library home pages included a contact phone number, a contact email address, an Ask-a-Librarian feature, the physical address listed, a feedback/suggestions link, subject guides, a discovery tool or database-specific search box, multimedia, social media, a site search option, a responsive web design, and a copyright year or update date.

  16. "I didn't know her, but…": parasocial mourning of mediated deaths on Facebook RIP pages

    NASA Astrophysics Data System (ADS)

    Klastrup, Lisbeth

    2015-04-01

    This article examines the use of six Danish "Rest in Peace" or (RIP) memorial pages. The article focuses on the relation between news media and RIP page use, in relation to general communicative practices on these pages. Based on an analysis of press coverage of the deaths of six young people and a close analysis of 1,015 comments extracted from the RIP pages created to memorialize them, it is shown that their deaths attracted considerable media attention, as did the RIP pages themselves. Comment activity seem to reflect the news stories in the way the commenters refer to the context of death and the emotional distress they experience, but mainly comments on the RIP pages are conventional expressions of sympathy and "RIP" wishes. The article concludes that public RIP pages might be understood as virtual spontaneous shrines, affording an emerging practice of "RIP-ing."

  17. Science on the Web: Secondary School Students' Navigation Patterns and Preferred Pages' Characteristics

    NASA Astrophysics Data System (ADS)

    Dimopoulos, Kostas; Asimakopoulos, Apostolos

    2010-06-01

    This study aims to explore navigation patterns and preferred pages' characteristics of ten secondary school students' searching the web for information about cloning. The students navigated the Web for as long as they wished in a context of minimum support of teaching staff. Their navigation patterns were analyzed using audit trail data software. The characteristics of their preferred Web pages were also analyzed using a scheme of analysis largely based on socio-linguistics and socio-semiotics approaches. Two distinct groups of students could be discerned. The first consisted of more competent students, who during their navigation visited fewer relevant pages, however of higher credibility and more specialized content. The second group consists of weaker students, who visited more pages, mainly of lower credibility and rather popularized content. Implications for designing educational web pages and teaching are discussed.

  18. CD and 31P NMR studies of tachykinin and MSH neuropeptides in SDS and DPC micelles

    NASA Astrophysics Data System (ADS)

    Schneider, Sydney C.; Brown, Taylor C.; Gonzalez, Javier D.; Levonyak, Nicholas S.; Rush, Lydia A.; Cremeens, Matthew E.

    2016-02-01

    Secondary structural characteristics of substance P (SP), neurokinin A (NKA), neurokinin B (NKB), α-melanocyte stimulating hormone peptide (α-MSH), γ1-MSH, γ2-MSH, and melittin were evaluated with circular dichroism in phosphite buffer, DPC micelles, and SDS micelles. CD spectral properties of γ1-MSH and γ2-MSH as well as 31P NMR of DPC micelles with all the peptides are reported for the first time. Although, a trend in the neuropeptide/micelle CD data appears to show increased α-helix content for the tachykinin peptides (SP, NKA, NKB) and increased β-sheet content for the MSH peptides (α-MSH, γ1-MSH, γ2-MSH) with increasing peptide charge, the lack of perturbed 31P NMR signals for all neuropeptides could suggest that the reported antimicrobial activity of SP and α-MSH might not be related to a membrane disruption mode of action.

  19. Characterization of major allergens of royal jelly Apis mellifera.

    PubMed

    Rosmilah, M; Shahnaz, M; Patel, G; Lock, J; Rahman, D; Masita, A; Noormalin, A

    2008-12-01

    Royal jelly is widely consumed in the community and has perceived benefits ranging from promoting growth in children and improvement of general health status to enhancement of longevity for the elderly. However, royal jelly consumption has been linked to contact dermatitis, acute asthma, anaphylaxis and death. High prevalence of positive skin tests to royal jelly have been reported among atopic populations in countries with a high rate of royal jelly consumption. The present study is aimed to identify the major allergens of royal jelly. Royal jelly extract was separated by sodium dodecyl polyacrylamide gel electrophoresis (SDS-PAGE) and 2-dimensional electrophoresis (2-D). Immunoblotting of the SDS-PAGE and 2-D profiles were performed to identify the allergenic spots. Spots were then excised from the 2-D gel, digested with trypsin and analyzed by mass spectrometry. The SDS-PAGE of royal jelly extract revealed 18 bands between 10 to 167 kD. Western blot of the fractionated proteins detected 15 IgE-binding bands between 14 to 127 kD with seven major allergens of 32, 40, 42, 49, 55, 60 and 67 kD using serum from 53 subjects with royal jelly allergy. The 2-D gel fractionated the royal jelly proteins to more than 50 different protein spots. Out of these, 30 spots demonstrated specific IgE affinity to the sera tested. Eight spots of the major royal jelly allergens were selected for mass-spectrometry analysis. Digested tryptic peptides of the spots were compared to the amino acid sequence search in protein databases which identified the fragments of royal jelly homologus to major royal jelly protein 1 (MRJ1) and major royal jelly protein 2 (MRJ2). In conclusion, the major allergens of royal jelly are MRJ1 and MRJ2 in our patients' population.

  20. Fractionation and identification of the allergic proteins in Aspergillus species.

    PubMed

    Falahati, M; Ghanbari, S; Ebrahimi, M; Ghazanfari, M; Bazrafshan, F; Farahyar, S; Falak, R

    2016-12-01

    Allergy is an undesired immune response to non-pathogenic agents. However, some opportunistic microorganisms such as fungi can also cause allergy. Among those fungi, hyphae form of Aspergillus strains including A. fumigatus , A. flavus , and A. niger could be mentioned. In this study, we aimed to separate allergic proteins from Aspergillus strains and determine their identity. Standard species of Aspergillus strains were cultivated in optimized conditions and the mycelium was separated by centrifugation. The fungal cells were lysed through physical methods such as freeze-thawing and grinding to prepare a suitable protein extract. The protein concentration was measured by Bradford method and the electrophoretic pattern of the extract was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The proteins were fractionated by ammonium sulfate precipitation and anion exchange chromatography using fast protein liquid chromatography (FPLC) system. The IgE immunoreactivity of the sensitized patients and controls was studied using the fractionated proteins by enzyme-linked immunosorbent assay (ELISA). Following SDS-PAGE, proteins were electrotransferred onto polyvinylidene difluoride (PVDF) membranes and the strips were blotted with allergic patients' and controls' sera. The immunoreactive bands were excised from colloidal coomassie-stained SDS-PAGE gels and studied by mass spectroscopy methods. Among the studied species, A. fumigatus showed stronger IgE reactivity and more IgE reactive protein bands than others did. The proteins with higher molecular weights showed stronger immunoreactivity in Western blotting. Receiver operating characteristic curve analysis demonstrated a correlation between the results of the applied ELISA methods. One of the most prominent IgE-reactive proteins was confirmed to be 45 kDa mycelia catalase. Our findings confirmed that high molecular weight proteins might play a major role in allergy and IgE reactivity to

  1. Native Mutant Huntingtin in Human Brain

    PubMed Central

    Sapp, Ellen; Valencia, Antonio; Li, Xueyi; Aronin, Neil; Kegel, Kimberly B.; Vonsattel, Jean-Paul; Young, Anne B.; Wexler, Nancy; DiFiglia, Marian

    2012-01-01

    Huntington disease (HD) is caused by polyglutamine expansion in the N terminus of huntingtin (htt). Analysis of human postmortem brain lysates by SDS-PAGE and Western blot reveals htt as full-length and fragmented. Here we used Blue Native PAGE (BNP) and Western blots to study native htt in human postmortem brain. Antisera against htt detected a single band broadly migrating at 575–850 kDa in control brain and at 650–885 kDa in heterozygous and Venezuelan homozygous HD brains. Anti-polyglutamine antisera detected full-length mutant htt in HD brain. There was little htt cleavage even if lysates were pretreated with trypsin, indicating a property of native htt to resist protease cleavage. A soluble mutant htt fragment of about 180 kDa was detected with anti-htt antibody Ab1 (htt-(1–17)) and increased when lysates were treated with denaturants (SDS, 8 m urea, DTT, or trypsin) before BNP. Wild-type htt was more resistant to denaturants. Based on migration of in vitro translated htt fragments, the 180-kDa segment terminated ≈htt 670–880 amino acids. If second dimension SDS-PAGE followed BNP, the 180-kDa mutant htt was absent, and 43–50 kDa htt fragments appeared. Brain lysates from two HD mouse models expressed native full-length htt; a mutant fragment formed if lysates were pretreated with 8 m urea + DTT. Native full-length mutant htt in embryonic HD140Q/140Q mouse primary neurons was intact during cell death and when cell lysates were exposed to denaturants before BNP. Thus, native mutant htt occurs in brain and primary neurons as a soluble full-length monomer. PMID:22375012

  2. Flagellin diversity in Clostridium botulinum groups I and II: a new strategy for strain identification.

    PubMed

    Paul, Catherine J; Twine, Susan M; Tam, Kevin J; Mullen, James A; Kelly, John F; Austin, John W; Logan, Susan M

    2007-05-01

    Strains of Clostridium botulinum are traditionally identified by botulinum neurotoxin type; however, identification of an additional target for typing would improve differentiation. Isolation of flagellar filaments and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that C. botulinum produced multiple flagellin proteins. Nano-liquid chromatography-tandem mass spectrometry (nLC-MS/MS) analysis of in-gel tryptic digests identified peptides in all flagellin bands that matched two homologous tandem flagellin genes identified in the C. botulinum Hall A genome. Designated flaA1 and flaA2, these open reading frames encode the major structural flagellins of C. botulinum. Colony PCR and sequencing of flaA1/A2 variable regions classified 80 environmental and clinical strains into group I or group II and clustered isolates into 12 flagellar types. Flagellar type was distinct from neurotoxin type, and epidemiologically related isolates clustered together. Sequencing a larger PCR product, obtained during amplification of flaA1/A2 from type E strain Bennett identified a second flagellin gene, flaB. LC-MS analysis confirmed that flaB encoded a large type E-specific flagellin protein, and the predicted molecular mass for FlaB matched that observed by SDS-PAGE. In contrast, the molecular mass of FlaA was 2 to 12 kDa larger than the mass predicted by the flaA1/A2 sequence of a given strain, suggesting that FlaA is posttranslationally modified. While identification of FlaB, and the observation by SDS-PAGE of different masses of the FlaA proteins, showed the flagellin proteins of C. botulinum to be diverse, the presence of the flaA1/A2 gene in all strains examined facilitates single locus sequence typing of C. botulinum using the flagellin variable region.

  3. Flagellin Diversity in Clostridium botulinum Groups I and II: a New Strategy for Strain Identification▿

    PubMed Central

    Paul, Catherine J.; Twine, Susan M.; Tam, Kevin J.; Mullen, James A.; Kelly, John F.; Austin, John W.; Logan, Susan M.

    2007-01-01

    Strains of Clostridium botulinum are traditionally identified by botulinum neurotoxin type; however, identification of an additional target for typing would improve differentiation. Isolation of flagellar filaments and analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed that C. botulinum produced multiple flagellin proteins. Nano-liquid chromatography-tandem mass spectrometry (nLC-MS/MS) analysis of in-gel tryptic digests identified peptides in all flagellin bands that matched two homologous tandem flagellin genes identified in the C. botulinum Hall A genome. Designated flaA1 and flaA2, these open reading frames encode the major structural flagellins of C. botulinum. Colony PCR and sequencing of flaA1/A2 variable regions classified 80 environmental and clinical strains into group I or group II and clustered isolates into 12 flagellar types. Flagellar type was distinct from neurotoxin type, and epidemiologically related isolates clustered together. Sequencing a larger PCR product, obtained during amplification of flaA1/A2 from type E strain Bennett identified a second flagellin gene, flaB. LC-MS analysis confirmed that flaB encoded a large type E-specific flagellin protein, and the predicted molecular mass for FlaB matched that observed by SDS-PAGE. In contrast, the molecular mass of FlaA was 2 to 12 kDa larger than the mass predicted by the flaA1/A2 sequence of a given strain, suggesting that FlaA is posttranslationally modified. While identification of FlaB, and the observation by SDS-PAGE of different masses of the FlaA proteins, showed the flagellin proteins of C. botulinum to be diverse, the presence of the flaA1/A2 gene in all strains examined facilitates single locus sequence typing of C. botulinum using the flagellin variable region. PMID:17351097

  4. Analysis of Soluble Proteins in Natural Cordyceps sinensis from Different Producing Areas by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis and Two-dimensional Electrophoresis

    PubMed Central

    Li, Chun-Hong; Zuo, Hua-Li; Zhang, Qian; Wang, Feng-Qin; Hu, Yuan-Jia; Qian, Zheng-Ming; Li, Wen-Jia; Xia, Zhi-Ning; Yang, Feng-Qing

    2017-01-01

    Background: As one of the bioactive components in Cordyceps sinensis (CS), proteins were rarely used as index components to study the correlation between the protein components and producing areas of natural CS. Objective: Protein components of 26 natural CS samples produced in Qinghai, Tibet, and Sichuan provinces were analyzed and compared to investigate the relationship among 26 different producing areas. Materials and Methods: Proteins from 26 different producing areas were extracted by Tris-HCl buffer with Triton X-100, and separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional electrophoresis (2-DE). Results: The SDS-PAGE results indicated that the number of protein bands and optical density curves of proteins in 26 CS samples was a bit different. However, the 2-DE results showed that the numbers and abundance of protein spots in protein profiles of 26 samples were obviously different and showed certain association with producing areas. Conclusions: Based on the expression values of matched protein spots, 26 batches of CS samples can be divided into two main categories (Tibet and Qinghai) by hierarchical cluster analysis. SUMMARY The number of protein bands and optical density curves of proteins in 26 Cordyceps sinensis samples were a bit different on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein profilesNumbers and abundance of protein spots in protein profiles of 26 samples were obvious different on two-dimensional electrophoresis mapsTwenty-six different producing areas of natural Cordyceps sinensis samples were divided into two main categories (Tibet and Qinghai) by Hierarchical cluster analysis based on the values of matched protein spots. Abbreviations Used: SDS-PAGE: Sodium dodecyl sulfate polyacrylamide gel electrophoresis, 2-DE: Two-dimensional electrophoresis, Cordyceps sinensis: CS, TCMs: Traditional Chinese medicines PMID:28250651

  5. [Protein interaction site of Toxoplasma gondii microneme protein 6 and aldolase determined by site-directed mutagenesis].

    PubMed

    Zheng, Bin; Yin, Zhi-Kui; Zhan, Xi-Mei

    2014-06-01

    To identify the protein interaction site of Toxoplasma gondii microneme protein 6 (MIC6) and aldolase by using site-directed mutagenesis. Based on Toxoplasma gondii MIC6 gene sequence (GenBank Accession No. AF110270), the specific primers were designed. Tryptophan (W)-348 of MIC6 C terminus (MIC6C) was mutated to valine (V) via site-directed mutagenesis. MIC6C W/V gene was obtained from cDNA library by PCR amplification and subcloned into pGEX-4T-1. The mutant protein GST-MIC6C W/V was expressed in E. coli, induced by 0.8 mmol/L IPTG, and purified by affinity chromatography. Glutathione sepharose beads were incubated with GST-MIC6C W/V and GST-MIC6C, respectively, and then incubated with T. gondii tachyzoites lysate, and bound proteins were eluted using sample buffer. Bound products were resolved by SDS-PAGE and Western blotting. Glutathione sepharose beads were incubated with GST-MIC6C W/V and GST-MIC6C, respectively, and then incubated with aldolase-His6. After incubation, the resin was washed and subjected to SDS-PAGE. The MIC6C W/N gene was obtained, and the recombinant plasmid MIC6C W/V/pGEX-4T-1 was successfully constructed. The mutant protein GST-MIC6C W/V was expressed and purified in vitro. SDS-PAGE analysis indicated that GST-MIC6C was co-precipitated with aldolase from T. gondii tachyzoites lysate or aldolase-His6, whereas GST-MIC6C W/V failed to precipitate aldolase from T. gondii tachyzoites lysate or aldolase-His6. Western blotting analysis using anti-aldolase antibody indicated that GST-MIC6C could pull-down aldolase from T. gondii tachyzoites lysate. Tryptophan (W348) was the interaction site of MIC6 and aldolase in T. gondii.

  6. Carnosine ameliorates lens protein turbidity formations by inhibiting calpain proteolysis and ultraviolet C-induced degradation.

    PubMed

    Liao, Jiahn-Haur; Lin, I-Lin; Huang, Kai-Fa; Kuo, Pei-Ting; Wu, Shih-Hsiung; Wu, Tzu-Hua

    2014-06-25

    Carnosine (CAR) is an endogenous peptide and present in lens, but there is little evidence for its effectiveness in calpain-induced proteolysis inhibition and its differential effects toward different wavelengths of ultraviolet (UV) irradiation. This study aimed to develop three in vitro cataract models to compare the mechanisms underlying the protective activities of CAR. Crude crystallins extracted from porcine lenses were used for antiproteolysis assays, and purified γ-crystallins were used for anti-UV assays. The turbidity in those in vitro models mimics cataract formation and was assayed by measuring optical density (OD) at 405 nm. The effectiveness of CAR on calpain-induced proteolysis was studied at 37 and 58 °C. Patterns of proteins were then analyzed by SDS-PAGE. The turbidity was reduced significantly (p<0.05) at 60 min measurements with the increased concentration of CAR (10-300 mM). SDS-PAGE showed that the decreased intensities at both ∼28 and ∼30 kDa protein bands in heat-enhanced assays were ameliorated by CAR at ≥10 mM concentrations. In UV-B studies, CAR (200, 300 mM) reduced the turbidity of γ-crystallin significantly (p<0.05) at 6 h observations. The turbidity of samples containing γ-crystallins was ameliorated while incubated with CAR (100, 300 mM) significantly (p<0.05) following 4 h of exposure to UV-C. SDS-PAGE showed that the presence of CAR reduced UV-B-induced aggregation of γ-crystallins at ∼44 kDa and resulted in less loss of γ-crystallin following UV-C exposure. The result of modeling also suggests that CAR acts as an inhibitor of calpain. In conclusion, CAR protects lens proteins more readily by inhibiting proteolysis and UV-C-induced degradation than aggregation induced by UV-B irradiation.

  7. Generation and purification of recombinant fimbrillin from Porphyromonas (Bacteroides) gingivalis 381.

    PubMed Central

    Washington, O R; Deslauriers, M; Stevens, D P; Lyford, L K; Haque, S; Yan, Y; Flood, P M

    1993-01-01

    Fimbrillin is the major subunit protein of fimbriae from the human periodontal pathogen Porphyromonas (Bacteroides) gingivalis. We describe here the generation and initial characterization of recombinant fimbrillin (r-fimbrillin) isolated from P. gingivalis 381. A fragment of DNA encoding the gene for fimbrillin was generated by polymerase chain reaction and cloned into the expression vector pET11b. Plasmids containing the recombinant gene were transfected into Escherichia coli. Clones were selected on plates for ampicillin resistance and individually screened by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) for protein production after activation with IPTG (isopropyl-beta-D- thiogalactopyranoside). One clone, OW0.2, produced significant amounts of a 42-kDa protein after induction with IPTG. This clone contained the pET11b plasmid with a 1-kb insert that had sequence homology to the gene encoding fimbrillin. The majority of recombinant protein from clone OW0.2 was found in the cytoplasm within inclusion bodies. Protein aggregates were solubilized in 8 M urea, and SDS-PAGE analysis showed two major protein bands, one at 42 kDa and the other at 17 kDa. These two proteins coeluted from a DEAE-Sepharose column at 0.15 M NaCl and were reactive to rabbit antiserum to fimbrillin in a Western blot (immunoblot). A preparation giving a single protein band at 42 kDa in SDS-PAGE was obtained by size fractionation by using continuous-elution electrophoresis. Lymph node cells from animals immunized with either fimbrillin from P. gingivalis or r-fimbrillin showed antigen-specific proliferation to both P. gingivalis fimbrillin and r-fimbrillin in an in vitro recall assay. Therefore, it appears that r-fimbrillin is chemically, antigenically, and serologically identical to fimbrillin isolated from P. gingivalis 381. Images PMID:8094377

  8. Effect of laser soldering irradiation on covalent bonds of pure collagen.

    PubMed

    Constantinescu, Mihai A; Alfieri, Alex; Mihalache, George; Stuker, Florian; Ducray, Angélique; Seiler, Rolf W; Frenz, Martin; Reinert, Michael

    2007-03-01

    Laser tissue welding and soldering is being increasingly used in the clinical setting for defined surgical procedures. The exact induced changes responsible for tensile strength are not yet fully investigated. To further improve the strength of the bonding, a better understanding of the laser impact at the subcellular level is necessary. The goal of this study was to analyze whether the effect of laser irradiation on covalent bonding in pure collagen using irradiances typically applied for tissue soldering. Pure rabbit and equine type I collagen were subjected to laser irradiation. In the first part of the study, rabbit and equine collagen were compared using identical laser and irradiation settings. In the second part of the study, equine collagen was irradiated at increasing laser powers. Changes in covalent bonding were studied indirectly using the sodium dodecylsulfate polyacrylamide gel electrophoresis (SDS-PAGE) technique. Tensile strengths of soldered membranes were measured with a calibrated tensile force gauge. In the first experiment, no differences between the species-specific collagen bands were noted, and no changes in banding were found on SDS-PAGE after laser irradiation. In the second experiment, increasing laser irradiation power showed no effect on collagen banding in SDS-PAGE. Finally, the laser tissue soldering of pure collagen membranes showed virtually no determinable tensile strength. Laser irradiation of pure collagen at typical power settings and exposure times generally used in laser tissue soldering does not induce covalent bonding between collagen molecules. This is true for both rabbit and equine collagen proveniences. Furthermore, soldering of pure collagen membranes without additional cellular components does not achieve the typical tensile strength reported in native, cell-rich tissues. This study is a first step in a better understanding of laser impact at the molecular level and might prove useful in engineering of combined

  9. Geographically diverse Australian isolates of Melissococcus pluton exhibit minimal genotypic diversity by restriction endonuclease analysis.

    PubMed

    Djordjevic, S P; Smith, L A; Forbes, W A; Hornitzky, M A

    1999-04-15

    Melissococcus pluton, the causative agent of European foulbrood is an economically significant disease of honey bees (Apis mellifera) across most regions of the world and is prevalent throughout most states of Australia. 49 Isolates of M. pluton recovered from diseased colonies or honey samples in New South Wales, Queensland, South Australia, Tasmania and Victoria were compared using SDS-PAGE, Western immunoblotting and restriction endonuclease analyses. DNA profiles of all 49 geographically diverse isolates showed remarkably similar AluI profiles although four isolates (one each from Queensland, South Australia, New South Wales and Victoria) displayed minor profile variations compared to AluI patterns of all other isolates. DNA from a subset of the 49 Australian and three isolates from the United Kingdom were digested separately with the restriction endonucleases CfoI, RsaI and DraI. Restriction endonuclease fragment patterns generated using these enzymes were also similar although minor variations were noted. SDS-PAGE of whole cell proteins from 13 of the 49 isolates from different states of Australia, including the four isolates which displayed minor profile variations (AluI) produced indistinguishable patterns. Major immunoreactive proteins of approximate molecular masses of 21, 24, 28, 30, 36, 40, 44, 56, 60, 71, 79 and 95 kDa were observed in immunoblots of whole cell lysates of 22 of the 49 isolates and reacted with rabbit hyperimmune antibodies raised against M. pluton whole cells. Neither SDS-PAGE or immunoblotting was capable of distinguishing differences between geographically diverse isolates of M. pluton. Collectively these data confirm that Australian isolates of M. pluton are genetically homogeneous and that this species may be clonal. Plasmid DNA was not detected in whole cell DNA profiles of any isolate resolved using agarose gel electrophoresis.

  10. Qualitative variation of photolabelled benzodiazepine receptors in different species.

    PubMed

    Hebebrand, J; Friedl, W; Lentes, K U; Propping, P

    1986-01-01

    In order to examine whether species differences of benzodiazepine receptor subunits exist, we compared the fluorographic pattern of photoaffinity labelled subunits after SDS-PAGE in five species: fish, frog, chicken, mouse and calf. Each species showed a distinct pattern of specifically labelled proteins. We conclude that species variation of benzodiazepine receptor does indeed exist.

  11. The structure and dynamics of Nano Particles encapsulated by the SDS monolayer collapse at the water/TCE interface

    PubMed Central

    Shi, Wenxiong

    2016-01-01

    The super-saturated surfactant monolayer collapses with the nanoparticles (NPs) at the water/trichloroethylene (TCE) interface are investigated using molecular dynamics (MD) simulations. The results show that sodium alkyl sulfate (SDS) monolayer collapse is initiated by buckling and followed primarily by budding and the bud encapsulating the NPs and oil molecules. The developed bud detaches from the monolayer into a water phase and forms the swollen micelle emulsion with NPs and oil molecules. We investigate the wavelength of the initial budding and the theoretical description of the budding process. The wavelength of the monolayer increases with bending modulus. The energy barrier of the budding can be easily overcome by thermal fluctuation energy, which indicates that budding process proceeds rapidly. PMID:27853312

  12. Assessment of Free Dye in Solutions of Dual-Labeled Antibody Conjugates for In Vivo Molecular Imaging

    PubMed Central

    Aldrich, Melissa B.; Wang, XueJuan; Hart, Amy; Sampath, Lakshmi; Marshall, Milton V.; Sevick-Muraca, Eva M.

    2017-01-01

    PURPOSE Recent preclinical and clinical studies show dyes that excite and fluoresce in the near infrared range may be used for tracking and detecting disease targets in vivo. A method for quantifying free dye molecules in antibody conjugate preparations is required for agent batch release and for translation into the clinic. PROCEDURES Herein, we developed and validated a SDS-PAGE method to determine the percentage of free IRDye 800CW in (DTPA)n-trastuzumab—(IRDye 800)m conjugate sample preparations in which HPLC assessment of free dye was not possible. RESULTS The SDS-PAGE assay was accurate and valid for free IRDye 800CW amounts between 38 and 4 molar percent of total dye. Gel sample preparation reagent affected the specificity of the assay, and lower and upper limits of quantitation and detection were determined. CONCLUSION This method may be applicable to other near infrared dye-conjugated antibody-based imaging agents in which HPLC assessment of purity is not feasible. This validated method for quality assurance will facilitate the translation of dual-labeled antibody conjugates for nuclear and optical imaging. PMID:20458634

  13. Different analytical approaches in assessing antibacterial activity and the purity of commercial lysozyme preparations for dairy application.

    PubMed

    Brasca, Milena; Morandi, Stefano; Silvetti, Tiziana; Rosi, Veronica; Cattaneo, Stefano; Pellegrino, Luisa

    2013-05-21

    Hen egg-white lysozyme (LSZ) is currently used in the food industry to limit the proliferation of lactic acid bacteria spoilage in the production of wine and beer, and to inhibit butyric acid fermentation in hard and extra hard cheeses (late blowing) caused by the outgrowth of clostridial spores. The aim of this work was to evaluate how the enzyme activity in commercial preparations correlates to the enzyme concentration and can be affected by the presence of process-related impurities. Different analytical approaches, including turbidimetric assay, SDS-PAGE and HPLC were used to analyse 17 commercial preparations of LSZ marketed in different countries. The HPLC method adopted by ISO allowed the true LSZ concentration to be determined with accuracy. The turbidimetric assay was the most suitable method to evaluate LSZ activity, whereas SDS-PAGE allowed the presence of other egg proteins, which are potential allergens, to be detected. The analytical results showed that the purity of commercially available enzyme preparations can vary significantly, and evidenced the effectiveness of combining different analytical approaches in this type of control.

  14. Purification and biochemical characterization of glucose 6-phosphate dehydrogenase, 6-phosphogluconate dehydrogenase and glutathione reductase from rat lung and inhibition effects of some antibiotics.

    PubMed

    Adem, Sevki; Ciftci, Mehmet

    2016-12-01

    G6PD, 6PGD and GR have been purified separately in the single step from rat lung using 2', 5'-ADP Sepharose 4B affinity chromatography. The purified enzymes showed a single band on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weights of the enzymes were estimated to be 134 kDa for G6PD, 107 kDa for 6PGD and 121 kDa for GR by Sephadex G-150 gel filtration chromatography, and the subunit molecular weights was respectively found to be 66, 52 and 63 kDa by SDS-PAGE. Optimum pH, stable pH, optimum ionic strength, optimum temperature, KM and Vmax values for substrates were determined. Product inhibition studies were also performed. The enzymes were inhibited by levofloxacin, furosemide, ceftazidime, cefuroxime and gentamicin as in vitro with IC50 values in the range of 0.07-30.13 mM. In vivo studies demonstrated that lung GR was inhibited by furosemide and lung 6PGD was inhibited by levofloxacin.

  15. Protein and alkaloid patterns of the floral nectar in some solanaceous species.

    PubMed

    Kerchner, András; Darók, Judit; Bacskay, Ivett; Felinger, Attila; Jakab, Gábor; Farkas, Ágnes

    2015-09-01

    The family Solanaceae includes several melliferous plants, which tend to produce copious amounts of nectar. Floral nectar is a chemically complex aqueous solution, dominated by sugars, but minor components such as amino acids, proteins, flavonoids and alkaloids are present as well. This study aimed at analysing the protein and alkaloid profile of the nectar in seven solanaceous species. Proteins were examined with SDS-PAGE and alkaloids were analyzed with HPLC. The investigation of protein profile revealed significant differences in nectar-protein patterns not only between different plant genera, but also between the three Nicotiana species investigated. SDS-PAGE suggested the presence of several Nectarin proteins with antimicrobial activity in Nicotiana species. The nectar of all tobacco species contained the alkaloid nicotine, N. tabacum having the highest nicotine content. The nectar of Brugmansia suaveolens, Datura stramonium, Hyoscyamus niger and Lycium barbarum contained scopolamine, the highest content of which was measured in B. suaveolens. The alkaloid concentrations in the nectars of most solanaceous species investigated can cause deterrence in honeybees, and the nectar of N. rustica and N. tabacum can be considered toxic for honeybees.

  16. Relationship between the dough quality and content of specific glutenin proteins in wheat mill streams, and its application to making flour suitable for instant Chinese noodles.

    PubMed

    Yahata, Eriko; Maruyama-Funatsuki, Wakako; Nishio, Zenta; Yamamoto, Yoshihiko; Hanaoka, Akihiro; Sugiyama, Hisashi; Tanida, Masatoshi; Saruyama, Haruo

    2006-04-01

    The content of specific proteins such as high-molecular-weight glutenin subunits HMW-GS 5+10 and low-molecular-weight glutenin subunits LMW-GS KS2 in wheat mill streams of extra-strong Kachikei 33 wheat was quantified by SDS-PAGE and 2D-PAGE. The mill streams showed varied quantities of HMW-GS 5+10 (0.077 to 2.007 mg/g of mill stream), LMW-GS KS2 (0.018 to 0.586 mg/g of mill stream) and total protein (9.42% to 18.98%). The contents of these specific proteins in the mill streams were significantly correlated with the SDS sedimentation volume and the mixing properties, which are respective indices of specific loaf volume and dough strength. The contents of these specific glutenin proteins in the mill streams were therefore found to be significantly important for improving the dough quality suitable for bread and Chinese noodles. Accordingly, we present here the application of this information to the development of an effective method for producing mill streams with high quality and yield that are suitable for instant Chinese noodles.

  17. Comparative Biochemical and Functional Studies on a Branded Human Recombinant Factor VIIa and a Biosimilar Equivalent Product.

    PubMed

    Sadeghi, Nasiredin; Kahn, Daniel; Syed, Daneyal; Iqbal, Omer; Abro, Schuharazad; Eshraghi, Reza; Hoppensteadt, Debra; Fareed, Jawed

    2014-09-01

    Recombinant factor VIIa (rFVIIa; NovoSeven, Novo Nordisk, Copenhagen, Denmark) is used to control bleeding in patients with hemophilia. A generic version of FVIIa was developed by AryoGen (Tehran, Iran). This study compared the composition and functional activities of AryoSeven and NovoSeven. Each product was compared at equigravimetric (1 mg/mL) stock solution for protein content. The proteomic profile was obtained using surface-enhanced laser desorption ionization mass spectrometry. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis was carried out to determine the protein profile and Western blotting was performed using a polyclonal rabbit antihuman FVIIa antibody. The FVIIa-related antigen was also measured using a commercially available enzyme-linked immunosorbent assay method. Functional assay included the prothrombin time correction in FVII-deficient plasma. The protein content was comparable in 2 products and the mass spectra analysis showed a single peak at 50 kDa in all products. The SDS-PAGE and immunoblotting studies were comparable. Both products exhibited similar coagulant properties in different assays. © The Author(s) 2014.

  18. Phosphorylation of the budgerigar fledgling disease virus major capsid protein VP1

    NASA Technical Reports Server (NTRS)

    Haynes, J. I. 2nd; Consigli, R. A.; Spooner, B. S. (Principal Investigator)

    1992-01-01

    The structural proteins of the budgerigar fledgling disease virus, the first known nonmammalian polyomavirus, were analyzed by isoelectric focusing and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The major capsid protein VP1 was found to be composed of at least five distinct species having isoelectric points ranging from pH 6.45 to 5.85. By analogy with the murine polyomavirus, these species apparently result from different modifications of an initial translation product. Primary chicken embryo cells were infected in the presence of 32Pi to determine whether the virus structural proteins were modified by phosphorylation. SDS-PAGE of the purified virus structural proteins demonstrated that VP1 (along with both minor capsid proteins) was phosphorylated. Two-dimensional analysis of the radiolabeled virus showed phosphorylation of only the two most acidic isoelectric species of VP1, indicating that this posttranslational modification contributes to VP1 species heterogeneity. Phosphoamino acid analysis of 32P-labeled VP1 revealed that phosphoserine is the only phosphoamino acid present in the VP1 protein.

  19. Production and biochemical characterization of insecticidal enzymes from Aspergillus fumigatus toward Callosobruchus maculatus.

    PubMed

    Pereira, Jackeline L; Franco, Octávio L; Noronha, Eliane F

    2006-06-01

    In the present work, Aspergillus fumigatus is described as a higher producer of hydrolytic enzymes secreted in response to the presence of the Callosobruchus maculatus bruchid pest. This fungus was able to grow over cowpea weevil shells as a unique carbon source, secreting alkaline proteolytic and chitinolytic enzymes. Enzyme secretion in A. fumigatus was induced by both C. maculatus exoskeleton as well as commercial chitin, and alkaline proteolytic and chitinolytic activities were detected after 48 hours of growth. Furthermore, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed the production of specific proteins. Among them, two extracellular alkaline proteinases from culture enriched with C. maculatus exoskeleton were purified after chromatographic procedures using ion exchange and affinity columns. These proteins, named AP15 and AP30, had apparent molecular masses of 15,500 and 30,000 Da, respectively, as estimated by SDS-PAGE electrophoresis and mass spectrometry. AP30 was classified as a serine proteinase because it was inhibited by 5 mM: phenylmethylsulfonyl fluoride (100%) and 50 microM leupeptin (67.94%).

  20. Production of peptone from boso fish (Oxyeleotris marmorata) for bacterial growth medium

    NASA Astrophysics Data System (ADS)

    Priatni, S.; Kosasih, W.; Budiwati, T. A.; Ratnaningrum, D.

    2017-03-01

    Underutilized Oxyeleotris marmorata fish is abundant and widespread in Indonesia. The study aimed to use O. marmorata fish for peptone production using papain from dried latex of papaya fruit. The peptone was applied as nitrogen sources for bacterial growth. The resulted peptone was optimized at 50-65°C for 5-8 hr, using 0.1% of papain at pH 6.0. Characterization of peptone was based on the soluble protein content, N-amino content, % degree hydrolysis (DH), SDS PAGE profile and growth of bacteria Escherichia coli and Staphylococcus aureus. The results indicated that the optimum condition for hydrolysis was at 50°C for 7 hr (p < 0.05). Fish peptone soluble protein content was of 8.6 mg/mL, α-amino was 0.59%, and AN/TN 5.47%. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS PAGE) profile of peptone showed a major band with molecular weight between 17-28 kDa. Fish peptone effectiveness for E. coli and S. aureus growth was similar with commercial bacterial peptone.

  1. [Construction and expression of a eukaryotic expression vector containing human CR2-Fc fusion protein].

    PubMed

    Li, Xinxin; Wu, Zhihao; Zhang, Chuanfu; Jia, Leili; Song, Hongbin; Xu, Yuanyong

    2014-01-01

    To construct a eukaryotic expression vector containing human complement receptor 2 (CR2)-Fc and express the CR2-Fc fusion protein in Chinese hamster ovary (CHO) cells. The extracellular domain of human CR2 and IgG1 Fc were respectively amplified, ligated and inserted into the eukaryotic expression vector PCI-neo. After verified by restriction enzyme digestion and sequencing, the recombinant plasmid was transfected into CHO K1 cells. The ones with stable expression of the fusion protein were obtained by means of G418 selection. The expression of the CR2-Fc fusion protein was detected and confirmed by SDS-PAGE and Western blotting. Restriction enzyme digestion and sequencing demonstrated that the recombinant plasmid was valid. SDS-PAGE showed that relative molecular mass (Mr;) of the purified product was consistent with the expected value. Western blotting further proved the single band at the same position. We constructed the eukaryotic expression vector of CR2-Fc/PCI-neo successfully. The obtained fusion protein was active and can be used for the further study of the role in HIV control.

  2. DOE Office of Scientific and Technical Information (OSTI.GOV)

    Woodson, W.R.; Handa, A.K.

    Changes in proteins associated with senescence of the flowers of Hibiscus rosa-sinensis was studied using SDS-PAGE. Total extractable protein from petals decreased with senescence. Changes were noted in patterns of proteins from aging petals. Flower opening and senescence was associated with appearance and disappearance of several polypeptides. One new polypeptide with an apparent mw of 41 kd was first seen the day of flower opening and increased to over 9% of the total protein content of senescent petal tissue. Protein synthesis during aging was investigated by following uptake and incorporation of /sup 3/H-leucine into TCA-insoluble fraction of petal discs. Proteinmore » synthesis, as evidenced by the percent of label incorporated into the TCA-insoluble fraction, was greatest (32%) the day before flower opening. Senescent petal tissue incorporated 4% of label taken up into protein. Proteins were separated by SDS-PAGE and labelled polypeptides identified by fluorography. In presenescent petal tissue, radioactivity was distributed among several major polypeptides. In senescent tissue, much of the radioactivity was concentrated in the 41 kd polypeptide.« less

  3. 16 CFR 436.3 - Cover page.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... business address, telephone number, and, if applicable, email address and primary home page address. (c) A... Commission. You can contact the FTC at 1-877-FTC-HELP or by writing to the FTC at 600 Pennsylvania Avenue, NW...

  4. The next generation of capillary electrophoresis instruments: Performance of CE-SDS protein analysis.

    PubMed

    Kahle, Julia; Maul, Kai Jorrit; Wätzig, Hermann

    2018-01-01

    Over the last decade, capillary electrophoresis gained tremendous importance, because it became an indispensible tool for the quality control of biologics, e.g. therapeutic antibodies. Consequently, there has been a continuous development within the CE market. Microchip techniques have been established in the last years. Further trends are complete solutions for specific applications by the usage of reagent kits. Step by step instructions and facilitated handling of the instruments are becoming more common. This work focuses on the sized-based protein analysis with CE-SDS. The instruments CE 7100 by Agilent Technologies, LabChip® GXII Touch HT by PerkinElmer, Maurice S. by Protein Simple and PrinCE NextI870 by Prince Technologies have been evaluated, mainly analyzing protein mixtures of different molecular weights in long series. Published data of the PA 800 plus by SCIEX are also included in the tabled results. Precision, reliability, flexibility, and speed have been identified as the most important performance parameters, others such as resolution, sensitivity, linearity, ease of use and sustainability have also been considered. All tested instruments have shown an excellent performance. Depending on application and necessities, each user can find the most appropriate one. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Accidentes laborales asociados al desánimo de médicos SERUMS para laborar en el primer nivel de atención de Lima, Perú

    PubMed Central

    Mejia, Christian R.; Valladares-Garrido, Mario J.; Romero, Brian M.; Valladares-Garrido, Danai; Linares-Reyes, Edgardo

    2018-01-01

    Introducción la retención laboral es un tema de suma importancia, porque se requiere de profesionales en el primer nivel de atención (PNA). El objetivo fue determinar si los accidentes laborales se asociaron al desánimo de los médicos para trabajar en el PNA de Lima, Perú. Métodos estudio transversal analítico de datos secundarios de una base de datos generada de una encuesta a médicos que realizaron su Servicio Rural y Urbano-Marginal en Salud (SERUMS). Se incluyó solo a los médicos que manifestaron al inicio del SERUMS que podían trabajar en el PNA de Lima. Se definió como cambio de intención de trabajo en el PNA a los que finalizando el SERUMS refirieron que ya no deseaban laborar en Lima. Esto se asoció según si tuvieron un accidente laboral y se ajustó por otras variables. Resultados de los 124 médicos el 63% fueron hombres (78). La mediana de edad fue de 26 años (rango intercuartílico: 25-27 años). Después de su SERUMS, el 12% (15) manifestó que cambió su interés y que deseaba trabajar en la capital. En el análisis multivariado, haber tenido un accidente laboral disminuyó la frecuencia del cambio de intención de trabajo en el PNA (RPa: 0.28, IC 95%: 0.14-0.54, p < 0,001), ajustado por ocho variables. Conclusiones en un estudio previo los accidentes laborales disminuyeron la frecuencia de trabajar en provincias, pero nuestro estudio dice lo contrario, posiblemente por la percepción de que un trabajo en la capital del país permite estar más cerca de los servicios para ser atendido en caso de cualquier emergencia. PMID:29190859

  6. Preparing for Phase II: A Guide to the Pay/Personnel Administrative Support System (PASS) Source Data System (SDS) Site Preparation Process for PASS Field Managers.

    DTIC Science & Technology

    1981-06-01

    of standard reports are: 1. Activity Locator; 2. Report of Active Duty Obligations and Projected Rotation Date; and 3. Enlisted Personnel Advancement ... advancement in rate, etc.), currently used forms are listed and analyzed to determine how such transactions are processed under the existing system...to suport an SDS functional organization. It is emphasized that configuration design should consider the requirements which will be imposed under all

  7. Structural Insights into the PorK and PorN Components of the Porphyromonas gingivalis Type IX Secretion System.

    PubMed

    Gorasia, Dhana G; Veith, Paul D; Hanssen, Eric G; Glew, Michelle D; Sato, Keiko; Yukitake, Hideharu; Nakayama, Koji; Reynolds, Eric C

    2016-08-01

    The type IX secretion system (T9SS) has been recently discovered and is specific to Bacteroidetes species. Porphyromonas gingivalis, a keystone pathogen for periodontitis, utilizes the T9SS to transport many proteins including the gingipain virulence factors across the outer membrane and attach them to the cell surface via a sortase-like mechanism. At least 11 proteins have been identified as components of the T9SS including PorK, PorL, PorM, PorN and PorP, however the precise roles of most of these proteins have not been elucidated and the structural organization of these components is unknown. In this study, we purified PorK and PorN complexes from P. gingivalis and using electron microscopy we have shown that PorN and the PorK lipoprotein interact to form a 50 nm diameter ring-shaped structure containing approximately 32-36 subunits of each protein. The formation of these rings was dependent on both PorK and PorN, but was independent of PorL, PorM and PorP. PorL and PorM were found to form a separate stable complex. PorK and PorN were protected from proteinase K cleavage when present in undisrupted cells, but were rapidly degraded when the cells were lysed, which together with bioinformatic analyses suggests that these proteins are exposed in the periplasm and anchored to the outer membrane via the PorK lipid. Chemical cross-linking and mass spectrometry analyses confirmed the interaction between PorK and PorN and further revealed that they interact with the PG0189 outer membrane protein. Furthermore, we established that PorN was required for the stable expression of PorK, PorL and PorM. Collectively, these results suggest that the ring-shaped PorK/N complex may form part of the secretion channel of the T9SS. This is the first report showing the structural organization of any T9SS component.

  8. Structural Insights into the PorK and PorN Components of the Porphyromonas gingivalis Type IX Secretion System

    PubMed Central

    Gorasia, Dhana G.; Veith, Paul D.; Hanssen, Eric G.; Glew, Michelle D.; Sato, Keiko; Yukitake, Hideharu; Nakayama, Koji; Reynolds, Eric C.

    2016-01-01

    The type IX secretion system (T9SS) has been recently discovered and is specific to Bacteroidetes species. Porphyromonas gingivalis, a keystone pathogen for periodontitis, utilizes the T9SS to transport many proteins including the gingipain virulence factors across the outer membrane and attach them to the cell surface via a sortase-like mechanism. At least 11 proteins have been identified as components of the T9SS including PorK, PorL, PorM, PorN and PorP, however the precise roles of most of these proteins have not been elucidated and the structural organization of these components is unknown. In this study, we purified PorK and PorN complexes from P. gingivalis and using electron microscopy we have shown that PorN and the PorK lipoprotein interact to form a 50 nm diameter ring-shaped structure containing approximately 32–36 subunits of each protein. The formation of these rings was dependent on both PorK and PorN, but was independent of PorL, PorM and PorP. PorL and PorM were found to form a separate stable complex. PorK and PorN were protected from proteinase K cleavage when present in undisrupted cells, but were rapidly degraded when the cells were lysed, which together with bioinformatic analyses suggests that these proteins are exposed in the periplasm and anchored to the outer membrane via the PorK lipid. Chemical cross-linking and mass spectrometry analyses confirmed the interaction between PorK and PorN and further revealed that they interact with the PG0189 outer membrane protein. Furthermore, we established that PorN was required for the stable expression of PorK, PorL and PorM. Collectively, these results suggest that the ring-shaped PorK/N complex may form part of the secretion channel of the T9SS. This is the first report showing the structural organization of any T9SS component. PMID:27509186

  9. Decoding 2D-PAGE complex maps: relevance to proteomics.

    PubMed

    Pietrogrande, Maria Chiara; Marchetti, Nicola; Dondi, Francesco; Righetti, Pier Giorgio

    2006-03-20

    This review describes two mathematical approaches useful for decoding the complex signal of 2D-PAGE maps of protein mixtures. These methods are helpful for interpreting the large amount of data of each 2D-PAGE map by extracting all the analytical information hidden therein by spot overlapping. Here the basic theory and application to 2D-PAGE maps are reviewed: the means for extracting information from the experimental data and their relevance to proteomics are discussed. One method is based on the quantitative theory of statistical model of peak overlapping (SMO) using the spot experimental data (intensity and spatial coordinates). The second method is based on the study of the 2D-autocovariance function (2D-ACVF) computed on the experimental digitised map. They are two independent methods that are able to extract equal and complementary information from the 2D-PAGE map. Both methods permit to obtain fundamental information on the sample complexity and the separation performance and to single out ordered patterns present in spot positions: the availability of two independent procedures to compute the same separation parameters is a powerful tool to estimate the reliability of the obtained results. The SMO procedure is an unique tool to quantitatively estimate the degree of spot overlapping present in the map, while the 2D-ACVF method is particularly powerful in simply singling out the presence of order in the spot position from the complexity of the whole 2D map, i.e., spot trains. The procedures were validated by extensive numerical computation on computer-generated maps describing experimental 2D-PAGE gels of protein mixtures. Their applicability to real samples was tested on reference maps obtained from literature sources. The review describes the most relevant information for proteomics: sample complexity, separation performance, overlapping extent, identification of spot trains related to post-translational modifications (PTMs).

  10. Mixed hemimicelles solid-phase extraction based on sodium dodecyl sulfate (SDS)-coated nano-magnets for the spectrophotometric determination of Fingolomid in biological fluids

    NASA Astrophysics Data System (ADS)

    Azari, Zhila; Pourbasheer, Eslam; Beheshti, Abolghasem

    2016-01-01

    In this study, mixed hemimicelles solid-phase extraction (SPE) based on sodium dodecyl sulfate (SDS)-coated nano-magnets Fe3O4 was investigated as a novel method for the separation and determination of Fingolimod (FLM) in water, urine and plasma samples prior to spectrophotometeric determination. Due to the high surface area of these new sorbents and the excellent adsorption capacity after surface modification by SDS, satisfactory extraction recoveries can be produced. The main factors affecting the adsolubilization of analysts, such as pH, surfactant and adsorbent amounts, ionic strength, extraction time and desorption conditions were studied and optimized. Under the selected conditions, FLM has been quantitatively extracted. The accuracy of the method was evaluated by recovery measurements on spiked samples, and good recoveries of 96%, 95% and 88% were observed for water, urine and plasma respectively. Proper linear behaviors over the investigated concentration ranges of 2-26, 2-17 and 2-13 mg/L with good coefficients of determination, 0.998, 0.997 and 0.995 were achieved for water, urine and plasma samples, respectively. To the best of our knowledge, this is the first time that a mixed hemimicelles SPE method based on magnetic separation and nanoparticles has been used as a simple and sensitive method for monitoring of FLM in water and biological samples.

  11. Social Responsibility and Corporate Web Pages: Self-Presentation or Agenda-Setting?

    ERIC Educational Resources Information Center

    Esrock, Stuart L.; Leichty, Greg B.

    1998-01-01

    Examines how corporate entities use the Web to present themselves as socially responsible citizens and to advance policy positions. Samples randomly "Fortune 500" companies, revealing that, although 90% had Web pages and 82% of the sites addressed a corporate social responsibility issue, few corporations used their pages to monitor…

  12. Expression of recombinant AccMRJP1 protein from royal jelly of Chinese honeybee in Pichia pastoris and its proliferation activity in an insect cell line

    USDA-ARS?s Scientific Manuscript database

    Main royal jelly protein 1 (MRJP1) is the most abundant member of the main royal jelly protein (MRJP) family among honeybees. Mature MRJP1 cDNA of the Chinese honeybee (Apis cerana cerana MRJP1, or AccMRJP1) was expressed in Pichia pastoris. SDS-PAGE showed that recombinant AccMRJP1 was identical in...

  13. Yellow pages advertising by physicians. Are doctors providing the information consumers want most?

    PubMed

    Butler, D D; Abernethy, A M

    1996-01-01

    Yellow pages listing are the most widely used form of physician advertising. Every month, approximately 21.6 million adults in the United States refer to the yellow pages before obtaining medical care. Mobile consumers--approximately 17% of the U.S. population who move each year--are heavy users of yellow pages. Consumers desire information on a physician's experience, but it is included in less than 1% of all physician display ads.

  14. Reconfigurable Full-Page Braille Displays

    NASA Technical Reports Server (NTRS)

    Garner, H. Douglas

    1994-01-01

    Electrically actuated braille display cells of proposed type arrayed together to form full-page braille displays. Like other braille display cells, these provide changeable patterns of bumps driven by digitally recorded text stored on magnetic tapes or in solid-state electronic memories. Proposed cells contain electrorheological fluid. Viscosity of such fluid increases in strong electrostatic field.

  15. Default Parallels Plesk Panel Page

    Science.gov Websites

    services that small businesses want and need. Our software includes key building blocks of cloud service virtualized servers Service Provider Products Parallels® Automation Hosting, SaaS, and cloud computing , the leading hosting automation software. You see this page because there is no Web site at this

  16. Next Linear Collider Home Page

    Science.gov Websites

    Welcome to the Next Linear Collider NLC Home Page If you would like to learn about linear colliders in general and about this next-generation linear collider project's mission, design ideas, and Linear Collider. line | NLC Home | NLC Technical | SLAC | mcdunn Tuesday, February 14, 2006 01:32:11 PM

  17. Aggregation study in mixture surfactant system TX-100+SDS in heavy water solutions by SANS method

    NASA Astrophysics Data System (ADS)

    Rajewska, A.; Islamov, A. Kh.; Bakeeva, R. F.

    2018-03-01

    The mixing of amphiphiles in water may lead to the formation of mixed micelles which often present new properties with respect to the pure component solutions [1,2]. The mixture system of classic surfactants SDS (sodium dodecyl sulfate)+TX-100(p-(1,1,3,3- tetramethyl) poly(oxyethylene) (anionic + non-ionic) in heavy water solutions was investigated at temperatures 30°, 50°, 70°C for compositions 1:1, 2:1, 3:1 by the small-angle neutron scattering(SANS) method on spectrometer (‘YuMO’) at the IBR-2 pulsed neutron source at FLNP, JINR in Dubna (Russia). Measurements have covered Q range from 8x10-3 to 0.4 Å-1. From the measured dependence of the scattered intensity on the scattering angle, we derived the size, shape of micelles, aggregation number at various compositions and temperatures. The size of mixed micelle is a weak function of the mixing ratio between the two components.

  18. Microseismic Event Relocation and Focal Mechanism Estimation Based on PageRank Linkage

    NASA Astrophysics Data System (ADS)

    Aguiar, A. C.; Myers, S. C.

    2017-12-01

    Microseismicity associated with enhanced geothermal systems (EGS) is key in understanding how subsurface stimulation can modify stress, fracture rock, and increase permeability. Large numbers of microseismic events are commonly associated with hydroshearing an EGS, making data mining methods useful in their analysis. We focus on PageRank, originally developed as Google's search engine, and subsequently adapted for use in seismology to detect low-frequency earthquakes by linking events directly and indirectly through cross-correlation (Aguiar and Beroza, 2014). We expand on this application by using PageRank to define signal-correlation topology for micro-earthquakes from the Newberry Volcano EGS in Central Oregon, which has been stimulated two times using high-pressure fluid injection. We create PageRank signal families from both data sets and compare these to the spatial and temporal proximity of associated earthquakes. PageRank families are relocated using differential travel times measured by waveform cross-correlation (CC) and the Bayesloc approach (Myers et al., 2007). Prior to relocation events are loosely clustered with events at a distance from the cluster. After relocation, event families are found to be tightly clustered. Indirect linkage of signals using PageRank is a reliable way to increase the number of events confidently determined to be similar, suggesting an efficient and effective grouping of earthquakes with similar physical characteristics (ie. location, focal mechanism, stress drop). We further explore the possibility of using PageRank families to identify events with similar relative phase polarities and estimate focal mechanisms following Shelly et al. (2016) method, where CC measurements are used to determine individual polarities within event clusters. Given a positive result, PageRank might be a useful tool in adaptive approaches to enhance production at well-instrumented geothermal sites. Prepared by LLNL under Contract DE-AC52-07NA27344

  19. Early zygote-specific nuclease in mitochondria of the true slime mold Physarum polycephalum.

    PubMed

    Moriyama, Yohsuke; Yamazaki, Tomokazu; Nomura, Hideo; Sasaki, Narie; Kawano, Shigeyuki

    2005-11-01

    The active, selective digestion of mtDNA from one parent is a possible molecular mechanism for the uniparental inheritance of mtDNA. In Physarum polycephalum, mtDNA is packed by DNA-binding protein Glom, which packs mtDNA into rod-shaped mt-nucleoids. After the mating, mtDNA from one parent is selectively digested, and the Glom began to disperse. Dispersed Glom was retained for at least 6 h after mtDNA digestion, but disappeared completely by about 12 h after mixing two strains. We identified two novel nucleases using DNA zymography with native-PAGE and SDS-PAGE. One is a Ca2+-dependent, high-molecular-weight nuclease complex (about 670 kDa), and the other is a Mn2+-dependent, high-molecular-weight nuclease complex (440-670 kDa); the activity of the latter was detected as a Mn2+-dependent, 13-kDa DNase band on SDS-PAGE. All mitochondria isolated from myxamoebae had mt-nucleoids, whereas half of the mitochondria isolated from the zygotes at 12 h after mixing had lost the mt-nucleoids. The activity of the Mn2+-dependent nuclease in the isolated mitochondria was detected at least 8 h after mixing of two strains. The timing and localization of the Mn2+-dependent DNase activity matched the selective digestion of mtDNA.

  20. Purification and biochemical characterization of a thermostable extracellular glucoamylase produced by the thermotolerant fungus Paecilomyces variotii.

    PubMed

    Michelin, Michele; Ruller, Roberto; Ward, Richard J; Moraes, Luiz Alberto B; Jorge, João A; Terenzi, Héctor F; Polizeli, Maria de Lourdes T M

    2008-01-01

    An extracellular glucoamylase produced by Paecilomyces variotii was purified using DEAE-cellulose ion exchange chromatography and Sephadex G-100 gel filtration. The purified protein migrated as a single band in 7% PAGE and 8% SDS-PAGE. The estimated molecular mass was 86.5 kDa (SDS-PAGE). Optima of temperature and pH were 55 degrees C and 5.0, respectively. In the absence of substrate the purified glucoamylase was stable for 1 h at 50 and 55 degrees C, with a t (50) of 45 min at 60 degrees C. The substrate contributed to protect the enzyme against thermal denaturation. The enzyme was mainly activated by manganese metal ions. The glucoamylase produced by P. variotii preferentially hydrolyzed amylopectin, glycogen and starch, and to a lesser extent malto-oligossacarides and amylose. Sucrose, p-nitrophenyl alpha-D-maltoside, methyl-alpha-D-glucopyranoside, pullulan, alpha- and beta-cyclodextrin, and trehalose were not hydrolyzed. After 24 h, the products of starch hydrolysis, analyzed by thin layer chromatography, showed only glucose. The circular dichroism spectrum showed a protein rich in alpha-helix. The sequence of amino acids of the purified enzyme VVTDSFR appears similar to glucoamylases purified from Talaromyces emersonii and with the precursor of the glucoamylase from Aspergillus oryzae. These results suggested the character of the enzyme studied as a glucoamylase (1,4-alpha-D-glucan glucohydrolase).

  1. Comparison of antimicrobial peptide purification via free-flow electrophoresis and gel filtration chromatography.

    PubMed

    Xia, Zhi-Jun; Liu, Zhen; Kong, Fan-Zhi; Fan, Liu-Yin; Xiao, Hua; Cao, Cheng-Xi

    2017-12-01

    Antimicrobial peptides (AMPs) are usually small and cationic biomolecules with broad-spectrum antimicrobial activities against pathogens. Purifying them from complex samples is essential to study their physiochemical properties. In this work, free-flow zone electrophoresis (FFZE) was utilized to purify AMPs from yeast fermentation broth. Meanwhile, gel filtration chromatography (GFC) was conducted for comparison. The separation efficiency was evaluated by SDS-PAGE analysis of the fractions from both methods. Our results demonstrated as follows: (i) FFZE had more than 30-fold higher processing capacity as compared with GFC; (ii) FFZE could achieve 87% purity and 89% recovery rate while in GFC these parameters were about 93 and 82%, respectively; (iii) the former had ∼2-fold dilution but the latter had ∼13-fold dilution. Furthermore, Tricine-SDS-PAGE, Native-PAGE, and gel IEF were carried out to characterize the purified AMPs. We found that two peptides existed as a pair with the molecular mass of ∼5.5 and 7.0 kDa, while the same pI 7.8. These two peptides were proved to have the antimicrobial activity through the standardized agar diffusion method. Therefore, FFZE could be used to continuously purify AMPs with high bioactivity, which will lead to its wide application in the clinical and pharmaceutical fields. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  2. The PorX Response Regulator of the Porphyromonas gingivalis PorXY Two-Component System Does Not Directly Regulate the Type IX Secretion Genes but Binds the PorL Subunit

    PubMed Central

    Vincent, Maxence S.; Durand, Eric; Cascales, Eric

    2016-01-01

    The Type IX secretion system (T9SS) is a versatile multi-protein complex restricted to bacteria of the Bacteriodetes phylum and responsible for the secretion or cell surface exposition of diverse proteins that participate to S-layer formation, gliding motility or pathogenesis. The T9SS is poorly characterized but a number of proteins involved in the assembly of the secretion apparatus in the oral pathogen Porphyromonas gingivalis have been identified based on genome substractive analyses. Among these proteins, PorY, and PorX encode typical two-component system (TCS) sensor and CheY-like response regulator respectively. Although the porX and porY genes do not localize at the same genetic locus, it has been proposed that PorXY form a bona fide TCS. Deletion of porX in P. gingivalis causes a slight decrease of the expression of a number of other T9SS genes, including sov, porT, porP, porK, porL, porM, porN, and porY. Here, we show that PorX and the soluble cytoplasmic domain of PorY interact. Using electrophoretic mobility shift, DNA-protein co-purification and heterologous host expression assays, we demonstrate that PorX does not bind T9SS gene promoters and does not directly regulate expression of the T9SS genes. Finally, we show that PorX interacts with the cytoplasmic domain of PorL, a component of the T9SS membrane core complex and propose that the CheY-like PorX protein might be involved in the dynamics of the T9SS. PMID:27630829

  3. Discovering urban mobility patterns with PageRank based traffic modeling and prediction

    NASA Astrophysics Data System (ADS)

    Wang, Minjie; Yang, Su; Sun, Yi; Gao, Jun

    2017-11-01

    Urban transportation system can be viewed as complex network with time-varying traffic flows as links to connect adjacent regions as networked nodes. By computing urban traffic evolution on such temporal complex network with PageRank, it is found that for most regions, there exists a linear relation between the traffic congestion measure at present time and the PageRank value of the last time. Since the PageRank measure of a region does result from the mutual interactions of the whole network, it implies that the traffic state of a local region does not evolve independently but is affected by the evolution of the whole network. As a result, the PageRank values can act as signatures in predicting upcoming traffic congestions. We observe the aforementioned laws experimentally based on the trajectory data of 12000 taxies in Beijing city for one month.

  4. Use of simulated pages to prepare medical students for internship and improve patient safety.

    PubMed

    Schwind, Cathy J; Boehler, Margaret L; Markwell, Stephen J; Williams, Reed G; Brenner, Michael J

    2011-01-01

    During the transition from medical school to internship, trainees experience high levels of stress related to pages on the inpatient wards. The steep learning curve during this period may also affect patient safety. The authors piloted the use of simulated pages to improve medical student preparedness, decrease stress related to pages, and familiarize medical students with common patient problems. A multidisciplinary team at Southern Illinois University School of Medicine developed simulated pages that were tested among senior medical students. Sixteen medical students were presented with 11 common patient scenarios. Data on assessment, management, and global performance were collected. Mean confidence levels were evaluated pre- and postintervention. Students were also surveyed on how the simulated pages program influenced their perceived comfort in managing patient care needs and the usefulness of the exercise in preparing them to handle inpatient pages. Mean scores on the assessment and management portions of the scenarios varied widely depending on the scenario (range -15.6 ± 41.6 to 95.7 ± 9.5). Pass rates based on global performance ranged from 12% to 93%. Interrater agreement was high (mean kappa = 0.88). Students' confidence ratings on a six-point scale increased from 1.87 preintervention to 3.53 postintervention (P < .0001). Simulated pages engage medical students and may foster medical student preparedness for internship. Students valued the opportunity to simulate "on call" responsibilities, and exposure to simulated pages significantly increased their confidence levels. Further studies are needed to determine effects on patient safety outcomes.

  5. Personal Web home pages of adolescents with cancer: self-presentation, information dissemination, and interpersonal connection.

    PubMed

    Suzuki, Lalita K; Beale, Ivan L

    2006-01-01

    The content of personal Web home pages created by adolescents with cancer is a new source of information about this population of potential benefit to oncology nurses and psychologists. Individual Internet elements found on 21 home pages created by youths with cancer (14-22 years old) were rated for cancer-related self-presentation, information dissemination, and interpersonal connection. Examples of adolescents' online narratives were also recorded. Adolescents with cancer used various Internet elements on their home pages for cancer-related self-presentation (eg, welcome messages, essays, personal history and diary pages, news articles, and poetry), information dissemination (e.g., through personal interest pages, multimedia presentations, lists, charts, and hyperlinks), and interpersonal connection (eg, guestbook entries). Results suggest that various elements found on personal home pages are being used by a limited number of young patients with cancer for self-expression, information access, and contact with peers.

  6. Comparison of quality of internet pages on human papillomavirus immunization in Italian and in English.

    PubMed

    Tozzi, Alberto Eugenio; Buonuomo, Paola Sabrina; Ciofi degli Atti, Marta Luisa; Carloni, Emanuela; Meloni, Marco; Gamba, Fiorenza

    2010-01-01

    Information available on the Internet about immunizations may influence parents' perception about human papillomavirus (HPV) immunization and their attitude toward vaccinating their daughters. We hypothesized that the quality of information on HPV available on the Internet may vary with language and with the level of knowledge of parents. To this end we compared the quality of a sample of Web pages in Italian with a sample of Web pages in English. Five reviewers assessed the quality of Web pages retrieved with popular search engines using criteria adapted from the Good Information Practice Essential Criteria for Vaccine Safety Web Sites recommended by the World Health Organization. Quality of Web pages was assessed in the domains of accessibility, credibility, content, and design. Scores in these domains were compared through nonparametric statistical tests. We retrieved and reviewed 74 Web sites in Italian and 117 in English. Most retrieved Web pages (33.5%) were from private agencies. Median scores were higher in Web pages in English compared with those in Italian in the domain of accessibility (p < .01), credibility (p < .01), and content (p < .01). The highest credibility and content scores were those of Web pages from governmental agencies or universities. Accessibility scores were positively associated with content scores (p < .01) and with credibility scores (p < .01). A total of 16.2% of Web pages in Italian opposed HPV immunization compared with 6.0% of those in English (p < .05). Quality of information and number of Web pages opposing HPV immunization may vary with the Web site language. High-quality Web pages on HPV, especially from public health agencies and universities, should be easily accessible and retrievable with common Web search engines. Copyright 2010 Society for Adolescent Medicine. Published by Elsevier Inc. All rights reserved.

  7. Accounting Programs' Home Pages: What's Happening.

    ERIC Educational Resources Information Center

    Peek, Lucia E.; Roxas, Maria L.

    2002-01-01

    Content analysis of 62 accounting programs' websites indicated the following: 53% include mission statements; 62.9% list accreditation; many faculty biographies and personal pages used inconsistent formats; provision of information on financial aid, student organizations, career services, and certified public accountant requirements varied. Many…

  8. Kinetic Stability of Proteins in Beans and Peas: Implications for Protein Digestibility, Seed Germination, and Plant Adaptation.

    PubMed

    Xia, Ke; Pittelli, Sandy; Church, Jennifer; Colón, Wilfredo

    2016-10-12

    Kinetically stable proteins (KSPs) are resistant to the denaturing detergent sodium dodecyl sulfate (SDS). Such resilience makes KSPs resistant to proteolytic degradation and may have arisen in nature as a mechanism for organismal adaptation and survival against harsh conditions. Legumes are well-known for possessing degradation-resistant proteins that often diminish their nutritional value. Here we applied diagonal two-dimensional (D2D) SDS-polyacrylamide gel electrophoresis (PAGE), a method that allows for the proteomics-level identification of KSPs, to a group of 12 legumes (mostly beans and peas) of agricultural and nutritional importance. Our proteomics results show beans that are more difficult to digest, such as soybean, lima beans, and various common beans, have high contents of KSPs. In contrast, mung bean, red lentil, and various peas that are highly digestible contain low amounts of KSPs. Identified proteins with high kinetic stability are associated with warm-season beans, which germinate at higher temperatures. In contrast, peas and red lentil, which are cool-season legumes, contain low levels of KSPs. Thus, our results show protein kinetic stability is an important factor in the digestibility of legume proteins and may relate to nutrition efficiency, timing of seed germination, and legume resistance to biotic stressors. Furthermore, we show D2D SDS-PAGE is a powerful method that could be applied for determining the abundance and identity of KSPs in engineered and wild legumes and for advancing basic research and associated applications.

  9. Identification and Characterization of Memecylon Species Using Isozyme Profiling

    PubMed Central

    Bharathi, T. R.; Sekhar, Shailasree; Geetha, N.; Niranjana, S. R.; Prakash, H. S.

    2017-01-01

    Background: The protein/isozyme fingerprint is useful in differentiating the species and acts as a biochemical marker for identification and systematic studies of medicinal plant species. Objective: In the present study, protein and isozyme profiles for peroxidase, esterase, acid phosphatase, polyphenol oxidase, alcohol dehydrogenase, and alkaline phosphatase of five species of Memecylon (Melastomataceae), Memecylon umbellatum, Memecylon edule, Memecylon talbotianum, Memecylon malabaricum, and Memecylon wightii were investigated. Materials and Methods: Fresh leaves were used to prepare crude enzyme extract for analyzing the five enzymes isozyme variations. Separation of isozymes was carried out using polyacrylamide gel electrophoresis (PAGE) and the banding patterns of protein were scored. Pair-wise comparisons of genotypes, based on the presence or absence of unique and shared polymorphic products, were used to regenerate similarity coefficients. The similarity coefficients were then used to construct dendrograms, using the unweighted pair group method with arithmetic averages. Results: A total of 50 bands with various Rf values and molecular weight were obtained through PAGE analysis. Among the five Memecylon species, more number of bands was produced in M. wightii and less number of bands was observed in M. edule. The results of similarity indices grouped M. malabaricum and M. wightii in one cluster with 98% similarity and M. umbellatum, M. edule, and M. talbotianum are grouped in another cluster with 79% similarity showing close genetic similarities which is in accordance with the morphological identification of Memecylon species. Conclusion: The protein/isozyme fingerprint is useful in differentiating the species and acts as a biochemical marker for identification of Memecylon species. SUMMARY Biochemical characterization of Memecylon species was evaluated by SDS-PAGE of extracted protein and isozyme profiling on native PAGE.After electrophoresis, each gel was

  10. Variability of protein content in calcium oxalate monohydrate stones.

    PubMed

    Williams, James C; Zarse, Chad A; Jackson, Molly E; Witzmann, Frank A; McAteer, James A

    2006-08-01

    Urinary stones are heterogeneous in their fragility to lithotripter shockwaves. As a first step in gaining a better understanding of the role of matrix in stone fragility, we measured extractible protein in calcium oxalate monohydrate (COM) stones that were extensively characterized by micro-computed tomography (micro CT). Stones were scanned using micro CT (Scanco mCT20, 34 microm). They were ground, and the protein extracted using four methods: 0.25M EDTA, 2% SDS reducing buffer, 9M urea buffer, and 10% acetic acid. Protein was measured using NanoOrange. The SDS extracts were also examined using polyacrylamide electrophoresis (PAGE). Extracted protein was highest with the SDS or urea methods (0.28% +/- 0.13% and 0.24% +/- 0.11%, respectively) and lower using the EDTA method (0.17% +/- 0.05%; P < 0.02). Acetic acid extracted little protein (0.006 +/- 0.002%; P < 0.001). Individual stones were significantly different in extractability of protein by the different methods, and SDS-PAGE revealed different protein patterns for individual stones. Extracted protein did not correlate with X-ray-lucent void percentage, which ranged from 0.06% to 2.8% of stone volume, or with apatite content. Extractible stone-matrix protein differs for individual COM stones, and yield is dependent on the extraction method. The presence of X-ray-lucent voids or minor amounts of apatite in stones did not correlate with protein content. The amounts of protein recovered were much lower than reported by Boyce, showing that these methods extracted only a fraction of the protein bound up in the stones. The results suggest that none of the methods tested will be useful for helping to answer the question of whether matrix content differs among stones of differing fragility to lithotripter shockwaves.

  11. Using Firefly Tools to Enhance Archive Web Pages

    NASA Astrophysics Data System (ADS)

    Roby, W.; Wu, X.; Ly, L.; Goldina, T.

    2013-10-01

    Astronomy web developers are looking for fast and powerful HTML 5/AJAX tools to enhance their web archives. We are exploring ways to make this easier for the developer. How could you have a full FITS visualizer or a Web 2.0 table that supports paging, sorting, and filtering in your web page in 10 minutes? Can it be done without even installing any software or maintaining a server? Firefly is a powerful, configurable system for building web-based user interfaces to access astronomy science archives. It has been in production for the past three years. Recently, we have made some of the advanced components available through very simple JavaScript calls. This allows a web developer, without any significant knowledge of Firefly, to have FITS visualizers, advanced table display, and spectrum plots on their web pages with minimal learning curve. Because we use cross-site JSONP, installing a server is not necessary. Web sites that use these tools can be created in minutes. Firefly was created in IRSA, the NASA/IPAC Infrared Science Archive (http://irsa.ipac.caltech.edu). We are using Firefly to serve many projects including Spitzer, Planck, WISE, PTF, LSST and others.

  12. 47 CFR 22.511 - Construction period for the Paging and Radiotelephone Service.

    Code of Federal Regulations, 2014 CFR

    2014-10-01

    ... 47 Telecommunication 2 2014-10-01 2014-10-01 false Construction period for the Paging and Radiotelephone Service. 22.511 Section 22.511 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES PUBLIC MOBILE SERVICES Paging and Radiotelephone Service § 22.511 Construction...

  13. 47 CFR 22.511 - Construction period for the Paging and Radiotelephone Service.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... 47 Telecommunication 2 2010-10-01 2010-10-01 false Construction period for the Paging and Radiotelephone Service. 22.511 Section 22.511 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES PUBLIC MOBILE SERVICES Paging and Radiotelephone Service § 22.511 Construction...

  14. 47 CFR 22.511 - Construction period for the Paging and Radiotelephone Service.

    Code of Federal Regulations, 2013 CFR

    2013-10-01

    ... 47 Telecommunication 2 2013-10-01 2013-10-01 false Construction period for the Paging and Radiotelephone Service. 22.511 Section 22.511 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES PUBLIC MOBILE SERVICES Paging and Radiotelephone Service § 22.511 Construction...

  15. 47 CFR 22.511 - Construction period for the Paging and Radiotelephone Service.

    Code of Federal Regulations, 2011 CFR

    2011-10-01

    ... 47 Telecommunication 2 2011-10-01 2011-10-01 false Construction period for the Paging and Radiotelephone Service. 22.511 Section 22.511 Telecommunication FEDERAL COMMUNICATIONS COMMISSION (CONTINUED) COMMON CARRIER SERVICES PUBLIC MOBILE SERVICES Paging and Radiotelephone Service § 22.511 Construction...

  16. Identification of Malicious Web Pages by Inductive Learning

    NASA Astrophysics Data System (ADS)

    Liu, Peishun; Wang, Xuefang

    Malicious web pages are an increasing threat to current computer systems in recent years. Traditional anti-virus techniques focus typically on detection of the static signatures of Malware and are ineffective against these new threats because they cannot deal with zero-day attacks. In this paper, a novel classification method for detecting malicious web pages is presented. This method is generalization and specialization of attack pattern based on inductive learning, which can be used for updating and expanding knowledge database. The attack pattern is established from an example and generalized by inductive learning, which can be used to detect unknown attacks whose behavior is similar to the example.

  17. Enriching the trustworthiness of health-related web pages.

    PubMed

    Gaudinat, Arnaud; Cruchet, Sarah; Boyer, Celia; Chrawdhry, Pravir

    2011-06-01

    We present an experimental mechanism for enriching web content with quality metadata. This mechanism is based on a simple and well-known initiative in the field of the health-related web, the HONcode. The Resource Description Framework (RDF) format and the Dublin Core Metadata Element Set were used to formalize these metadata. The model of trust proposed is based on a quality model for health-related web pages that has been tested in practice over a period of thirteen years. Our model has been explored in the context of a project to develop a research tool that automatically detects the occurrence of quality criteria in health-related web pages.

  18. Weather Prediction Center (WPC) Home Page

    Science.gov Websites

    grids, quantitative precipitation, and winter weather outlook probabilities can be found at: http Short Range Products » More Medium Range Products Quantitative Precipitation Forecasts Legacy Page Discussion (Day 1-3) Quantitative Precipitation Forecast Discussion NWS Weather Prediction Center College

  19. Referencing web pages and e-journals.

    PubMed

    Bryson, David

    2013-12-01

    One of the areas that can confuse students and authors alike is how to reference web pages and electronic journals (e-journals). The aim of this professional development article is to go back to first principles for referencing and see how with examples these should be referenced.

  20. WMO Sand and Dust Storm Warning Advisory and Assessment System (SDS-WAS): Research Implementation Status

    NASA Astrophysics Data System (ADS)

    Nickovic, Slobodan; Barrie, Leonard

    2010-05-01

    Assessment System (SDS-WAS) in order to improve the capabilities of countries affected by dust to reduce risks associated with airborne sand and dust. This project is in response to the desire of more than 40 WMO member countries to improve capabilities for more reliable sand and dust storm forecasts. The project has strong crosscutting features: it relies on real-time delivery of products; it integrates research communities (modelling, observation groups, and effects) and communities of practice (e.g. medical, aeronautical, agricultural users). There are two already established SDS-WAS nodes (Asian and North-Africa-Europe-Middle East) that coordinate implementation of the project objectives at regional levels. This presentation will review current status and future steps in the project implementation.

  1. Web pages: What can you see in a single fixation?

    PubMed

    Jahanian, Ali; Keshvari, Shaiyan; Rosenholtz, Ruth

    2018-01-01

    Research in human vision suggests that in a single fixation, humans can extract a significant amount of information from a natural scene, e.g. the semantic category, spatial layout, and object identities. This ability is useful, for example, for quickly determining location, navigating around obstacles, detecting threats, and guiding eye movements to gather more information. In this paper, we ask a new question: What can we see at a glance at a web page - an artificial yet complex "real world" stimulus? Is it possible to notice the type of website, or where the relevant elements are, with only a glimpse? We find that observers, fixating at the center of a web page shown for only 120 milliseconds, are well above chance at classifying the page into one of ten categories. Furthermore, this ability is supported in part by text that they can read at a glance. Users can also understand the spatial layout well enough to reliably localize the menu bar and to detect ads, even though the latter are often camouflaged among other graphical elements. We discuss the parallels between web page gist and scene gist, and the implications of our findings for both vision science and human-computer interaction.

  2. Watershed Data Management (WDM) database for West Branch DuPage River streamflow simulation, DuPage County, Illinois, January 1, 2007, through September 30, 2013

    USGS Publications Warehouse

    Bera, Maitreyee

    2017-10-16

    The U.S. Geological Survey (USGS), in cooperation with the DuPage County Stormwater Management Department, maintains a database of hourly meteorological and hydrologic data for use in a near real-time streamflow simulation system. This system is used in the management and operation of reservoirs and other flood-control structures in the West Branch DuPage River watershed in DuPage County, Illinois. The majority of the precipitation data are collected from a tipping-bucket rain-gage network located in and near DuPage County. The other meteorological data (air temperature, dewpoint temperature, wind speed, and solar radiation) are collected at Argonne National Laboratory in Argonne, Ill. Potential evapotranspiration is computed from the meteorological data using the computer program LXPET (Lamoreux Potential Evapotranspiration). The hydrologic data (water-surface elevation [stage] and discharge) are collected at U.S.Geological Survey streamflow-gaging stations in and around DuPage County. These data are stored in a Watershed Data Management (WDM) database.This report describes a version of the WDM database that is quality-assured and quality-controlled annually to ensure datasets are complete and accurate. This database is named WBDR13.WDM. It contains data from January 1, 2007, through September 30, 2013. Each precipitation dataset may have time periods of inaccurate data. This report describes the methods used to estimate the data for the periods of missing, erroneous, or snowfall-affected data and thereby improve the accuracy of these data. The other meteorological datasets are described in detail in Over and others (2010), and the hydrologic datasets in the database are fully described in the online USGS annual water data reports for Illinois (U.S. Geological Survey, 2016) and, therefore, are described in less detail than the precipitation datasets in this report.

  3. PageRank as a method to rank biomedical literature by importance.

    PubMed

    Yates, Elliot J; Dixon, Louise C

    2015-01-01

    Optimal ranking of literature importance is vital in overcoming article overload. Existing ranking methods are typically based on raw citation counts, giving a sum of 'inbound' links with no consideration of citation importance. PageRank, an algorithm originally developed for ranking webpages at the search engine, Google, could potentially be adapted to bibliometrics to quantify the relative importance weightings of a citation network. This article seeks to validate such an approach on the freely available, PubMed Central open access subset (PMC-OAS) of biomedical literature. On-demand cloud computing infrastructure was used to extract a citation network from over 600,000 full-text PMC-OAS articles. PageRanks and citation counts were calculated for each node in this network. PageRank is highly correlated with citation count (R = 0.905, P < 0.01) and we thus validate the former as a surrogate of literature importance. Furthermore, the algorithm can be run in trivial time on cheap, commodity cluster hardware, lowering the barrier of entry for resource-limited open access organisations. PageRank can be trivially computed on commodity cluster hardware and is linearly correlated with citation count. Given its putative benefits in quantifying relative importance, we suggest it may enrich the citation network, thereby overcoming the existing inadequacy of citation counts alone. We thus suggest PageRank as a feasible supplement to, or replacement of, existing bibliometric ranking methods.

  4. Does content affect whether users remember that Web pages were hyperlinked?

    PubMed

    Jones, Keith S; Ballew, Timothy V; Probst, C Adam

    2008-10-01

    We determined whether memory for hyperlinks improved when they represented relations between the contents of the Web pages. J. S. Farris (2003) found that memory for hyperlinks improved when they represented relations between the contents of the Web pages. However, Farris's (2003) participants could have used their knowledge of site content to answer questions about relations that were instantiated via the site's content and its hyperlinks. In Experiment 1, users navigated a Web site and then answered questions about relations that were instantiated only via content, only via hyperlinks, and via content and hyperlinks. Unlike Farris (2003), we split the latter into two sets. One asked whether certain content elements were related, and the other asked whether certain Web pages were hyperlinked. Experiment 2 replicated Experiment 1 with one modification: The questions that were asked about relations instantiated via content and hyperlinks were changed so that each question's wrong answer was also related to the question's target. Memory for hyperlinks improved when they represented relations instantiated within the content of the Web pages. This was true when (a) questions about content and hyperlinks were separated (Experiment 1) and (b) each question's wrong answer was also related to the question's target (Experiment 2). The accuracy of users' mental representations of local architecture depended on whether hyperlinks were related to the site's content. Designers who want users to remember hyperlinks should associate those hyperlinks with content that reflects the relation between the contents on the Web pages.

  5. Monitoring refolding of tailspike endorhamnosidase using capillary electrophoresis-laser induced tryptophan fluorescence

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Jensen, P.K.; Lee, Cheng S.; King, J.A.

    1997-12-31

    The use of capillary electrophoresis equipped with laser-induced tryptophan fluorescence detection is presented for monitoring the refolding pathway of phage P22 tailspike endorhamnosidase. Upon initiation of refolding, tailspike polypeptides rapidly fold into structured monomeric intermediates with a high content of secondary structure. These monomeric species associate to form the triple-chain defined folding intermediates, the protrimers. Conversion of the protrimer into the native, sodium dodecyl sulfate (SDS) resistant tailspike protein is the rate-limiting step in the refolding pathway. Refolding kinetics and yield measured by capillary electrophoresis are in good agreement with those obtained via native gel electrophoresis, SDS polyacrylamide gel electrophoresismore » (SDS-PAGE) and fluorescence spectrophotometry. To enhance separation resolution between protrimer and native protein in capillary electrophoresis, the use of poly(ethylene oxide) is investigated for the introduction of a sieving separation mechanism. The increased viscosity of the electrophoresis buffer may also play a role in resolution enhancement.« less

  6. Biophysical insights into the interaction of hen egg white lysozyme with therapeutic dye clofazimine: modulation of activity and SDS induced aggregation of model protein.

    PubMed

    Ajmal, Mohammad Rehan; Chaturvedi, Sumit Kumar; Zaidi, Nida; Alam, Parvez; Zaman, Masihuz; Siddiqi, Mohammad Khursheed; Nusrat, Saima; Jamal, Mohammad Sarwar; Mahmoud, Mohamed H; Badr, Gamal; Khan, Rizwan Hasan

    2017-08-01

    The present study details the binding process of clofazimine to hen egg white lysozyme (HEWL) using spectroscopy, dynamic light scattering, transmission electron microscopy (TEM), and molecular docking techniques. Clofazimine binds to the protein with binding constant (K b ) in the order of 1.57 × 10 4 at 298 K. Binding process is spontaneous and exothermic. Molecular docking results suggested the involvement of hydrogen bonding and hydrophobic interactions in the binding process. Bacterial cell lytic activity in the presence of clofazimine increased to more than 40% of the value obtained with HEWL only. Interaction of the drug with HEWL induced ordered secondary structure in the protein and molecular compaction. Clofazimine also effectively inhibited the sodium dodecyl sulfate (SDS) induced amyloid formation in HEWL and caused disaggregation of preformed fibrils, reinforcing the notion that there is involvement of hydrophobic interactions and hydrogen bonding in the binding process of clofazimine with HEWL and clofazimine destabilizes the mature fibrils. Further, TEM images confirmed that fibrillar species were absent in the samples where amyloid induction was performed in the presence of clofazimine. As clofazimine is a drug less explored for the inhibition of fibril formation of the proteins, this study reports the inhibition of SDS-induced amyloid formation of HEWL by clofazimine, which will help in the development of clofazimine-related molecules for the treatment of amyloidosis.

  7. Application-Controlled Demand Paging for Out-of-Core Visualization

    NASA Technical Reports Server (NTRS)

    Cox, Michael; Ellsworth, David; Kutler, Paul (Technical Monitor)

    1997-01-01

    In the area of scientific visualization, input data sets are often very large. In visualization of Computational Fluid Dynamics (CFD) in particular, input data sets today can surpass 100 Gbytes, and are expected to scale with the ability of supercomputers to generate them. Some visualization tools already partition large data sets into segments, and load appropriate segments as they are needed. However, this does not remove the problem for two reasons: 1) there are data sets for which even the individual segments are too large for the largest graphics workstations, 2) many practitioners do not have access to workstations with the memory capacity required to load even a segment, especially since the state-of-the-art visualization tools tend to be developed by researchers with much more powerful machines. When the size of the data that must be accessed is larger than the size of memory, some form of virtual memory is simply required. This may be by segmentation, paging, or by paged segments. In this paper we demonstrate that complete reliance on operating system virtual memory for out-of-core visualization leads to poor performance. We then describe a paged segment system that we have implemented, and explore the principles of memory management that can be employed by the application for out-of-core visualization. We show that application control over some of these can significantly improve performance. We show that sparse traversal can be exploited by loading only those data actually required. We show also that application control over data loading can be exploited by 1) loading data from alternative storage format (in particular 3-dimensional data stored in sub-cubes), 2) controlling the page size. Both of these techniques effectively reduce the total memory required by visualization at run-time. We also describe experiments we have done on remote out-of-core visualization (when pages are read by demand from remote disk) whose results are promising.

  8. Search Engine Ranking, Quality, and Content of Web Pages That Are Critical Versus Noncritical of Human Papillomavirus Vaccine.

    PubMed

    Fu, Linda Y; Zook, Kathleen; Spoehr-Labutta, Zachary; Hu, Pamela; Joseph, Jill G

    2016-01-01

    Online information can influence attitudes toward vaccination. The aim of the present study was to provide a systematic evaluation of the search engine ranking, quality, and content of Web pages that are critical versus noncritical of human papillomavirus (HPV) vaccination. We identified HPV vaccine-related Web pages with the Google search engine by entering 20 terms. We then assessed each Web page for critical versus noncritical bias and for the following quality indicators: authorship disclosure, source disclosure, attribution of at least one reference, currency, exclusion of testimonial accounts, and readability level less than ninth grade. We also determined Web page comprehensiveness in terms of mention of 14 HPV vaccine-relevant topics. Twenty searches yielded 116 unique Web pages. HPV vaccine-critical Web pages comprised roughly a third of the top, top 5- and top 10-ranking Web pages. The prevalence of HPV vaccine-critical Web pages was higher for queries that included term modifiers in addition to root terms. Compared with noncritical Web pages, Web pages critical of HPV vaccine overall had a lower quality score than those with a noncritical bias (p < .01) and covered fewer important HPV-related topics (p < .001). Critical Web pages required viewers to have higher reading skills, were less likely to include an author byline, and were more likely to include testimonial accounts. They also were more likely to raise unsubstantiated concerns about vaccination. Web pages critical of HPV vaccine may be frequently returned and highly ranked by search engine queries despite being of lower quality and less comprehensive than noncritical Web pages. Copyright © 2016 Society for Adolescent Health and Medicine. Published by Elsevier Inc. All rights reserved.

  9. Formal Features of Cyberspace: Relationships between Web Page Complexity and Site Traffic.

    ERIC Educational Resources Information Center

    Bucy, Erik P.; Lang, Annie; Potter, Robert F.; Grabe, Maria Elizabeth

    1999-01-01

    Examines differences between the formal features of commercial versus noncommercial Web sites, and the relationship between Web page complexity and amount of traffic a site receives. Findings indicate that, although most pages in this stage of the Web's development remain technologically simple and noninteractive, there are significant…

  10. College of DuPage Planning Process: College of DuPage Planning, the Foundation for Decision Making.

    ERIC Educational Resources Information Center

    College of DuPage, Glen Ellyn, IL. Office of Research and Planning.

    At College of DuPage (CD), in Glen Ellyn, Illinois, institutional planning is a multi-level, on-going process involving continuous college-wide input. The nine schematic levels in the CD planning process are interconnected and progress from global and broad-based issues, answering such questions as why the college exists, to concrete levels…

  11. An SDS-PAGE Examination of Protein Quaternary Structure and Disulfide Bonding for a Biochemistry Laboratory

    ERIC Educational Resources Information Center

    Powers, Jennifer L.; Andrews, Carla S.; St. Antoine, Caroline C.; Jain, Swapan S.; Bevilacqua, Vicky L. H.

    2005-01-01

    Electrophoresis is a valuable tool for biochemists, yet this technique is often not included in biochemistry laboratory curricula owing to time constraints or lack of equipment. Protein structure is also a topic of interest in many disciplines, yet most undergraduate lab experiments focus only on primary structure. In this experiment, students use…

  12. Comparison of lipopolysaccharides from Bacteroides, Porphyromonas, Prevotella, Campylobacter and Wolinella spp. by tricine-SDS-PAGE.

    PubMed

    Firoozkoohi, J; Zandi, H; Olsen, I

    1997-02-01

    Lipopolysaccharides (LPSs) of 11 bacterial strains from the type species of the genera Bacteroides (B. fragilis), Prevotella (Pr. melaninogenica), Porphyromonas (Po. gingivalis), Campylobacter (C. fetus subsp. fetus), and Wolinella (W. succinogenes), and from the type strains of B. distasonis, B. forsythus, B. ureolyticus, Po. levii, Po. macacae, and C. gracilis, were extracted with hot water-phenol (Westphal method). S-form LPSs, obtained from all organisms, were well resolved with tricine-sodium-dodecyl-sulphate polyacrylamide gel electrophoresis and visualized by silver staining. Lipid A was not stained. Also profiles from LPS of Escherichia coli, serotypes 0111:B4 and 055:B5, could be distinguished. While W. succinogenes showed a relatively short S-form LPS on electrophoregrams, the other bacteria, including B. fragilis, exhibited long-ladder LPSs. Po. gingivalis displayed the largest number of bands and the longest O-chain. The long O-chain of this bacterium may be important for its virulence.

  13. Intention to continue using Facebook fan pages from the perspective of social capital theory.

    PubMed

    Lin, Kuan-Yu; Lu, Hsi-Peng

    2011-10-01

    Social network sites enable users to express themselves, establish ties, and develop and maintain social relationships. Recently, many companies have begun using social media identity (e.g., Facebook fan pages) to enhance brand attractiveness, and social network sites have evolved into social utility networks, thereby creating a number of promising business opportunities. To this end, the operators of fan pages need to be aware of the factors motivating users to continue their patronization of such pages. This study set out to identify these motivating factors from the point of view of social capital. This study employed structural equation modeling to investigate a research model based on a survey of 327 fan pages users. This study discovered that ties related to social interaction (structural dimension), shared values (cognitive dimension), and trust (relational dimension) play important roles in users' continued intention to use Facebook fan pages. Finally, this study discusses the implications of these findings and offers directions for future research.

  14. Oncoprotein protein kinase

    DOEpatents

    Karin, Michael; Hibi, Masahiko; Lin, Anning

    1997-01-01

    An isolated polypeptide (JNK) characterized by having a molecular weight of 46kD as determined by reducing SDS-PAGE, having serine and threonine kinase activity, phosphorylating the c-Jun N-terminal activation domain and polynucleotide sequences and method of detection of JNK are provided herein. JNK phosphorylates c-Jun N-terminal activation domain which affects gene expression from AP-1 sites.

  15. Oncoprotein protein kinase antibody kit

    DOEpatents

    Karin, Michael [San Diego, CA; Hibi, Masahiko [San Diego, CA; Lin, Anning [La Jolla, CA

    2008-12-23

    An isolated polypeptide (JNK) characterized by having a molecular weight of 46 kD as determined by reducing SDS-PAGE, having serine and threonine kinase activity, phosphorylating the c-Jun N-terminal activation domain and polynucleotide sequences and method of detection of JNK are provided herein. JNK phosphorylates c-Jun N-terminal activation domain which affects gene expression from AP-1 sites.

  16. Oncoprotein protein kinase

    DOEpatents

    Karin, Michael; Hibi, Masahiko; Lin, Anning; Davis, Roger; Derijard, Benoit

    2003-02-04

    An isolated polypeptide (JNK) characterized by having a molecular weight of 46kD as determined by reducing SDS-PAGE, having serine and threonine kinase activity, phosphorylating the c-Jun N-terminal activation domain and polynucleotide sequences and method of detection of JNK are provided herein. JNK phosphorylates c-Jun N-terminal activation domain which affects gene expression from AP-1 sites.

  17. Oncoprotein protein kinase

    DOEpatents

    Karin, Michael; Hibi, Masahiko; Lin, Anning

    1997-01-01

    An isolated polypeptide (JNK) characterized by having a molecular weight of 46 kD as determined by reducing SDS-PAGE, having serine and threonine kinase activity, phosphorylating the c-Jun N-terminal activation domain and polynucleotide sequences and method of detection of JNK are provided herein. JNK phosphorylates c-Jun N-terminal activation domain which affects gene expression from AP-1 sites.

  18. Oncoprotein protein kinase

    DOEpatents

    Karin, Michael; Hibi, Masahiko; Lin, Anning

    1998-01-01

    An isolated polypeptide (JNK) characterized by having a molecular weight of 46 kD as determined by reducing SDS-PAGE, having serine and threonine kinase activity, phosphorylating the c-Jun N-terminal activation domain and polynucleotide sequences and method of detection of JNK are provided herein. JNK phosphorylates c-Jun N-terminal activation domain which affects gene expression from AP-1 sites.

  19. ARL Physics Web Pages: An Evaluation by Established, Transitional and Emerging Benchmarks.

    ERIC Educational Resources Information Center

    Duffy, Jane C.

    2002-01-01

    Provides an overview of characteristics among Association of Research Libraries (ARL) physics Web pages. Examines current academic Web literature and from that develops six benchmarks to measure physics Web pages: ease of navigation; logic of presentation; representation of all forms of information; engagement of the discipline; interactivity of…

  20. An Ant Colony Optimization Based Feature Selection for Web Page Classification

    PubMed Central

    2014-01-01

    The increased popularity of the web has caused the inclusion of huge amount of information to the web, and as a result of this explosive information growth, automated web page classification systems are needed to improve search engines' performance. Web pages have a large number of features such as HTML/XML tags, URLs, hyperlinks, and text contents that should be considered during an automated classification process. The aim of this study is to reduce the number of features to be used to improve runtime and accuracy of the classification of web pages. In this study, we used an ant colony optimization (ACO) algorithm to select the best features, and then we applied the well-known C4.5, naive Bayes, and k nearest neighbor classifiers to assign class labels to web pages. We used the WebKB and Conference datasets in our experiments, and we showed that using the ACO for feature selection improves both accuracy and runtime performance of classification. We also showed that the proposed ACO based algorithm can select better features with respect to the well-known information gain and chi square feature selection methods. PMID:25136678

  1. An ant colony optimization based feature selection for web page classification.

    PubMed

    Saraç, Esra; Özel, Selma Ayşe

    2014-01-01

    The increased popularity of the web has caused the inclusion of huge amount of information to the web, and as a result of this explosive information growth, automated web page classification systems are needed to improve search engines' performance. Web pages have a large number of features such as HTML/XML tags, URLs, hyperlinks, and text contents that should be considered during an automated classification process. The aim of this study is to reduce the number of features to be used to improve runtime and accuracy of the classification of web pages. In this study, we used an ant colony optimization (ACO) algorithm to select the best features, and then we applied the well-known C4.5, naive Bayes, and k nearest neighbor classifiers to assign class labels to web pages. We used the WebKB and Conference datasets in our experiments, and we showed that using the ACO for feature selection improves both accuracy and runtime performance of classification. We also showed that the proposed ACO based algorithm can select better features with respect to the well-known information gain and chi square feature selection methods.

  2. Environment: General; Grammar & Usage; Money Management; Music History; Web Page Creation & Design.

    ERIC Educational Resources Information Center

    Web Feet, 2001

    2001-01-01

    Describes Web site resources for elementary and secondary education in the topics of: environment, grammar, money management, music history, and Web page creation and design. Each entry includes an illustration of a sample page on the site and an indication of the grade levels for which it is appropriate. (AEF)

  3. Future Trends in Children's Web Pages: Probing Hidden Biases for Information Quality

    ERIC Educational Resources Information Center

    Kurubacak, Gulsun

    2007-01-01

    As global digital communication continues to flourish, Children's Web pages become more critical for children to realize not only the surface but also breadth and deeper meanings in presenting these milieus. These pages not only are very diverse and complex but also enable intense communication across social, cultural and political restrictions…

  4. Turning the Page with Preconference Workshops

    ERIC Educational Resources Information Center

    Knowledge Quest, 2011

    2011-01-01

    For those who are experiencing a lack of creative inspiration within their school library program but are ready to "turn a page" in their career or school library program, they may head to Minneapolis to attend one of the many great preconference workshops. This article presents and describes preconference workshops design to rid…

  5. 16 CFR 436.3 - Cover page.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... with a cover page, in the order and form as follows: (a) The title “FRANCHISE DISCLOSURE DOCUMENT” in... begin operation of a [franchise system name] franchise is [the total amount of Item 7 (§ 436.5(g... affiliate. (2) This disclosure document summarizes certain provisions of your franchise agreement and other...

  6. 16 CFR 436.3 - Cover page.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... with a cover page, in the order and form as follows: (a) The title “FRANCHISE DISCLOSURE DOCUMENT” in... begin operation of a [franchise system name] franchise is [the total amount of Item 7 (§ 436.5(g... affiliate. (2) This disclosure document summarizes certain provisions of your franchise agreement and other...

  7. 16 CFR 436.3 - Cover page.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... with a cover page, in the order and form as follows: (a) The title “FRANCHISE DISCLOSURE DOCUMENT” in... begin operation of a [franchise system name] franchise is [the total amount of Item 7 (§ 436.5(g... affiliate. (2) This disclosure document summarizes certain provisions of your franchise agreement and other...

  8. 16 CFR 436.3 - Cover page.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... with a cover page, in the order and form as follows: (a) The title “FRANCHISE DISCLOSURE DOCUMENT” in... begin operation of a [franchise system name] franchise is [the total amount of Item 7 (§ 436.5(g... affiliate. (2) This disclosure document summarizes certain provisions of your franchise agreement and other...

  9. Key-phrase based classification of public health web pages.

    PubMed

    Dolamic, Ljiljana; Boyer, Célia

    2013-01-01

    This paper describes and evaluates the public health web pages classification model based on key phrase extraction and matching. Easily extendible both in terms of new classes as well as the new language this method proves to be a good solution for text classification faced with the total lack of training data. To evaluate the proposed solution we have used a small collection of public health related web pages created by a double blind manual classification. Our experiments have shown that by choosing the adequate threshold value the desired value for either precision or recall can be achieved.

  10. Optimization of single plate-serial dilution spotting (SP-SDS) with sample anchoring as an assured method for bacterial and yeast cfu enumeration and single colony isolation from diverse samples.

    PubMed

    Thomas, Pious; Sekhar, Aparna C; Upreti, Reshmi; Mujawar, Mohammad M; Pasha, Sadiq S

    2015-12-01

    We propose a simple technique for bacterial and yeast cfu estimations from diverse samples with no prior idea of viable counts, designated as single plate-serial dilution spotting (SP-SDS) with the prime recommendation of sample anchoring (10 0 stocks). For pure cultures, serial dilutions were prepared from 0.1 OD (10 0 ) stock and 20 μl aliquots of six dilutions (10 1 -10 6 ) were applied as 10-15 micro-drops in six sectors over agar-gelled medium in 9-cm plates. For liquid samples 10 0 -10 5 dilutions, and for colloidal suspensions and solid samples (10% w/v), 10 1 -10 6 dilutions were used. Following incubation, at least one dilution level yielded 6-60 cfu per sector comparable to the standard method involving 100 μl samples. Tested on diverse bacteria, composite samples and Saccharomyces cerevisiae , SP-SDS offered wider applicability over alternative methods like drop-plating and track-dilution for cfu estimation, single colony isolation and culture purity testing, particularly suiting low resource settings.

  11. Trayectoria de los tornillos pediculares lumbares y sacros: Comparación entre el abordaje por linea media versus el abordaje posterolateral tipo wiltse

    PubMed Central

    Gagliardi, Martín; Guiroy, Alfredo; Molina, Federico Fernández; Fasano, Francisco; Ciancio, Alejandro Morales; Mezzadri, Juan José; Jalón, Pablo

    2017-01-01

    Resumen Objetivos: El objetivo de este estudio fue comparar, en fusiones lumbosacras cortas, el ángulo de convergencia de los tornillos pediculares entre el abordaje posterolateral tipo Wiltse y el abordaje mediano convencional. Método: Se revisaron en forma retrospectiva los controles en tomografía axial computada (TAC) de 76 tornillos pediculares lumbares y sacros colocados por vía posterior, mediante un abordaje mediano convencional (n: 38) o por vía posterolateral transmuscular tipo Wiltse (n: 38). Se incluyeron fusiones lumbosacras cortas desde L3 a S1, en pacientes adultos, con patología degenerativa. Se excluyeron los tornillos con una brecha ósea >4 mm en cualquier dirección, los casos con instrumentaciones pediculares previas y aquellos con curvas en el plano coronal mayores de 20°. Resultados: Considerando la totalidad de los implantes, el ángulo de convergencia fue de 23,3° (+/- 15,82). La angulación promedio, en el grupo AW, fue de 29,3° (+/- 9,72). En el grupo AC, el grado de convergencia de los implantes fue de 17,2° (+/- 10,58). Esta diferencia fue estadísticamente significativa (P < 0,0001). Para el grupo AW, el grado de convergencia según nivel fue el siguiente: L3: 31,2° (+/- 1,9); L4: 31,4° (+/- 2,76); L5: 31,1° (+/- 5,62); S1: 24,2° (+/- 12,16). El promedio del ángulo del tornillo según nivel para el grupo AC fue: L3: 16° (+/- 7,16); L4: 20,3° (+/- 6,9) L5: 15,9° (+/- 13,38); S1: 15,2° (+/- 14,32). Los implantes del grupo AW tuvieron ángulos significativamente más convergentes que el grupo AC en todos los segmentos explorados. Conclusión: En las fusiones lumbosacras cortas, la utilización del abordaje tipo Wiltse permitió la colocación de tornillos pediculares con más convergencia que en el abordaje mediano convencional. La relevancia clínica de este hecho es desconocida y se requerirían trabajos prospectivos randomizados para determinar la misma. PMID:29142777

  12. PagP Crystallized from SDS/Cosolvent Reveals the Route for Phospholipid Access to the Hydrocarbon Ruler

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Cuesta-Seijo, Jose Antonio; Neale, Chris; Khan, M. Adil

    2012-02-06

    Enzymatic reactions involving bilayer lipids occur in an environment with strict physical and topological constraints. The integral membrane enzyme PagP transfers a palmitoyl group from a phospholipid to lipid A in order to assist Escherichia coli in evading host immune defenses during infection. PagP measures the palmitoyl group with an internal hydrocarbon ruler that is formed in the interior of the eight-stranded antiparallel {beta} barrel. The access and egress of the palmitoyl group is thought to take a lateral route from the bilayer phase to the barrel interior. Molecular dynamics, mutagenesis, and a 1.4 {angstrom} crystal structure of PagP inmore » an SDS/2-methyl-2,4-pentanediol (MPD) cosolvent system reveal that phospholipid access occurs at the crenel present between strands F and G of PagP. In this way, the phospholipid head group can remain exposed to the cell exterior while the lipid acyl chain remains in a predominantly hydrophobic environment as it translocates to the protein interior.« less

  13. Analysis of the liver mitochondrial proteome in response to ethanol and S-adenosylmethionine treatments: novel molecular targets of disease and hepatoprotection.

    PubMed

    Andringa, Kelly K; King, Adrienne L; Eccleston, Heather B; Mantena, Sudheer K; Landar, Aimee; Jhala, Nirag C; Dickinson, Dale A; Squadrito, Giuseppe L; Bailey, Shannon M

    2010-05-01

    S-adenosylmethionine (SAM) minimizes alcohol hepatotoxicity; however, the molecular mechanisms responsible for SAM hepatoprotection remain unknown. Herein, we use proteomics to determine whether the hepatoprotective action of SAM against early-stage alcoholic liver disease is linked to alterations in the mitochondrial proteome. For this, male rats were fed control or ethanol-containing liquid diets +/- SAM and liver mitochondria were prepared for proteomic analysis. Two-dimensional isoelectric focusing (2D IEF/SDS-PAGE) and blue native gel electrophoresis (BN-PAGE) were used to determine changes in matrix and oxidative phosphorylation (OxPhos) proteins, respectively. SAM coadministration minimized alcohol-dependent inflammation and preserved mitochondrial respiration. SAM supplementation preserved liver SAM levels in ethanol-fed rats; however, mitochondrial SAM levels were increased by ethanol and SAM treatments. With use of 2D IEF/SDS-PAGE, 30 proteins showed significant changes in abundance in response to ethanol, SAM, or both. Classes of proteins affected by ethanol and SAM treatments were chaperones, beta oxidation proteins, sulfur metabolism proteins, and dehydrogenase enzymes involved in methionine, glycine, and choline metabolism. BN-PAGE revealed novel changes in the levels of 19 OxPhos proteins in response to ethanol, SAM, or both. Ethanol- and SAM-dependent alterations in the proteome were not linked to corresponding changes in gene expression. In conclusion, ethanol and SAM treatment led to multiple changes in the liver mitochondrial proteome. The protective effects of SAM against alcohol toxicity are mediated, in part, through maintenance of proteins involved in key mitochondrial energy conserving and biosynthetic pathways. This study demonstrates that SAM may be a promising candidate for treatment of alcoholic liver disease.

  14. Using Facebook Page Insights Data to Determine Posting Best Practices in an Academic Health Sciences Library

    ERIC Educational Resources Information Center

    Houk, Kathryn M.; Thornhill, Kate

    2013-01-01

    Tufts University Hirsh Health Sciences Library created a Facebook page and a corresponding managing committee in March 2010. Facebook Page Insights data collected from the library's Facebook page were statistically analyzed to investigate patterns of user engagement. The committee hoped to improve posting practices and increase user engagement…

  15. Ultra-processed food product brands on Facebook pages: highly accessed by Brazilians through their marketing techniques.

    PubMed

    Horta, Paula M; Rodrigues, Fernanda T; Dos Santos, Luana C

    2018-06-01

    To analyse the content and extent of marketing of ultra-processed food products (UPP) and their brand pages on Facebook, which are highly accessed by Brazilians. Descriptive. Sixteen UPP brand pages on Facebook were selected from 250 pages that were the most liked by Brazilians in October 2015. We analysed the frequency of 'likes' and members 'talking about' each one of the pages, in addition to fifteen marketing techniques used in the previous year (September 2014 to October 2015). The number of posts, likes, 'shares' and 'commentaries', and the mean number of likes, shares and commentaries per post, were collected for one month, from 23 September to 23 October 2015. The two most liked pages were: Coke® (93 673 979 likes) and McDonald's® (59 749 819 likes). Regarding the number of people talking about the pages, McDonald's led with 555 891 commentaries, followed by Coke (287 274), Burger King® (246 148) and Kibon® (244 523). All pages used marketing techniques, which included photos, user conversations, presence of brand elements and links. Videos were observed on 93·8 % of the pages; promotions on 68·8 %; and celebrities on 62·5 %. In one month, Garoto®, Outback® and Coke were brands that published more than one post per day. Kibon achieved the highest ratio of likes per post (285 845·50) and Burger King had the highest mean shares per post (10 083·93), including commentaries per post (7958·13). UPP marketing is extensively used on Facebook pages and is highly accessed by Brazilians, with UPP companies employing a diversity of marketing strategies.

  16. Facebook's personal page modelling and simulation

    NASA Astrophysics Data System (ADS)

    Sarlis, Apostolos S.; Sakas, Damianos P.; Vlachos, D. S.

    2015-02-01

    In this paper we will try to define the utility of Facebook's Personal Page marketing method. This tool that Facebook provides, is modelled and simulated using iThink in the context of a Facebook marketing agency. The paper has leveraged the system's dynamic paradigm to conduct Facebook marketing tools and methods modelling, using iThink™ system to implement them. It uses the design science research methodology for the proof of concept of the models and modelling processes. The following model has been developed for a social media marketing agent/company, Facebook platform oriented and tested in real circumstances. This model is finalized through a number of revisions and iterators of the design, development, simulation, testing and evaluation processes. The validity and usefulness of this Facebook marketing model for the day-to-day decision making are authenticated by the management of the company organization. Facebook's Personal Page method can be adjusted, depending on the situation, in order to maximize the total profit of the company which is to bring new customers, keep the interest of the old customers and deliver traffic to its website.

  17. Pre-labeling of diverse protein samples with a fixed amount of Cy5 for sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis.

    PubMed

    Bjerneld, Erik J; Johansson, Johan D; Laurin, Ylva; Hagner-McWhirter, Åsa; Rönn, Ola; Karlsson, Robert

    2015-09-01

    A pre-labeling protocol based on Cy5 N-hydroxysuccinimide (NHS) ester labeling of proteins has been developed for one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. We show that a fixed amount of sulfonated Cy5 can be used in the labeling reaction to label proteins over a broad concentration range-more than three orders of magnitude. The optimal amount of Cy5 was found to be 50 to 250pmol in 20μl using a Tris-HCl labeling buffer at pH 8.7. Labeling protein samples with a fixed amount of dye in this range balances the requirements of sub-nanogram detection sensitivity and low dye-to-protein (D/P) ratios for SDS-PAGE. Simulations of the labeling reaction reproduced experimental observations of both labeling kinetics and D/P ratios. Two-dimensional electrophoresis was used to examine the labeling of proteins in a cell lysate using both sulfonated and non-sulfonated Cy5. For both types of Cy5, we observed efficient labeling across a broad range of molecular weights and isoelectric points. Copyright © 2015 Elsevier Inc. All rights reserved.

  18. Using an FPLC to promote active learning of the principles of protein structure and purification.

    PubMed

    Robinson, Rebekah L; Neely, Amy E; Mojadedi, Wais; Threatt, Katie N; Davis, Nicole Y; Weiland, Mitch H

    2017-01-02

    The concepts of protein purification are often taught in undergraduate biology and biochemistry lectures and reinforced during laboratory exercises; however, very few reported activities allow students to directly gain experience using modern protein purification instruments, such as Fast Protein Liquid Chromatography (FPLC). This laboratory exercise uses size exclusion chromatography (SEC) and ion exchange (IEX) chromatography to separate a mixture of four different proteins. Students use an SEC chromatogram and corresponding SDS-PAGE gel to understand how protein conformations change under different conditions (i.e. native and non-native). Students explore strategies to separate co-eluting proteins by IEX chromatography. Using either cation or anion exchange, one protein is bound to the column while the other is collected in the flow-through. In this exercise, undergraduate students gain hands-on experience with experimental design, buffer and sample preparation, and implementation of instrumentation that is commonly used by experienced researchers while learning and applying the fundamental concepts of protein structure, protein purification, and SDS-PAGE. © 2016 by The International Union of Biochemistry and Molecular Biology, 45(1):60-68, 2017. © 2016 The International Union of Biochemistry and Molecular Biology.

  19. Simple, high-yield purification of xanthine oxidase from bovine milk.

    PubMed

    Ozer, N; Müftüoglu, M; Ataman, D; Ercan, A; Ogüs, I H

    1999-05-13

    Xanthine oxidase, a commercially important enzyme with a wide area of application, was extracted from fresh milk, without added preservatives, using toluene and heat. The short purification procedure, with high yield, consisted of extraction, ammonium sulfate fractionation, and DEAE-Sepharose (fast flow) column chromatography. Xanthine oxidase was eluted as a single activity peak from the column using a buffer gradient. The purification fold, specific activity and yield for the purified xanthine oxidase were 328, 10.161 U/mg and 69%, respectively. The enzyme was concentrated by ultrafiltration, although 31% of the activity was lost during concentration, no change in specific activity was observed. Activity and protein gave coincident staining bands on native polyacrylamide gels. The intensity and the number of bands were dependent on the oxidative state(s) of the enzyme; reduction by 2-mercaptoethanol decreased the intensity of the slow-moving bands and increased the intensity of the fastest-moving band. Following sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), two major bands (molecular masses of 152 and 131 kDa) were observed, accounting for > or = 95% of xanthine oxidase. Native- and SDS-PAGE showed that the purified xanthine oxidase becomes a heterodimer due to endogenous proteases.

  20. [The isolation and evaluation of Aspergillus fumigatus antigens].

    PubMed

    Lirio, V de S; de Assis, C M; Cano, M I; Lacaz, C da S

    1992-01-01

    Antigens from three strains of Aspergillus fumigatus (354, 356, and JIG) and an antiserum against the mixing of these antigens have been produced, and evaluated immunochemically. The antigens were obtained through a modified Coleman & Kaufman technique (culture filtrate concentrated by acetone). Analysis by the immunodiffusion test (ID) against homologous serum has yielded 100% sensitivity (with the studied sera). Concerning heterologous sera we found reactivity with a serum of a patient of candidiasis and another with histoplasmosis. The same result was obtained with a reference antigen in immunodiffusion, showing similar standards of response. Titration of the antiserum by ID and counterimmunoelectrophoresis showed a title of 1:32, and by complement fixation (micro-technique) a title of 1:128. Using immunoelectrophoresis (IEF), the produced antiserum yielded 8 lines of precipitation (5 in the anodic pole and 3 in the cathodic one). In SDS-PAGE at 12.5% the antigen has presented a rather complex electrophoretic profile (26 proteic subunits with a molecular weight ranging from 18 a > 100 kDa). Immunogenicity of the antigen was observed in all fractions of SDS-PAGE when the immunoblotting against the antiserum was carried out.

  1. Activity and Differential Induction of Chitinase Isozymes in Soybean Cultivars Resistant or Susceptible to Root-knot Nematodes.

    PubMed

    Qtu, J; Hallmann, J; Kokalis-Burelle, N; Weaver, D B; Rodríguez-Kábana, R; Tuzun, S

    1997-12-01

    Host physiological events in relation to infestation by parasitic nematodes are not well documented. Soybean plant responses to Meloidogyne incognita infestation were compared to resistant (Bryan) and susceptible (Brim) cultivars at 0, 1, 3, 10, 20, and 34 days after infestation (DAI). The resistant cultivar had higher chitinase activity than the susceptible cultivar at every sample time beginning at 3 DAI. Results from isoelectric focusing gel electrophoresis analyses indicated that three acidic chitinase isozymes with isoelectric points (pIs) of 4.8, 4.4, and 4.2 accumulated to a greater extent in the resistant compared to the susceptible cultivar following challenge. SDS-PAGE analysis of root proteins revealed that two proteins with molecular weights of approximately 31 and 46 kD accumulated more rapidly and to a higher level in the resistant than in the susceptible cultivar. Additionally, three major protein bands (33, 22, and 20 kD) with chitinase activity were detected with a modified SDS-PAGE analysis in which glycolchitin was added into the gel matrix. These results indicate that higher chitinase activity and early induction of specific chitinase isozymes may be associated with resistance to root-knot nematode in soybean.

  2. Studies on effect of additives on protein profile, microstructure and quality characteristics of pasta.

    PubMed

    Purnima, C; Ramasarma, P R; Prabhasankar, P

    2012-02-01

    Wheat storage proteins play a vital role in pasta making quality. In the present study, SDS-PAGE, Gel filtration chromatography and Scanning electron microscopy techniques were employed to understand the changes in the wheat protein fractions and their interactions with additives namely Sodium Steroyl Lactate (SSL), Glycerol Monostearate (GMS) and Hydroxy Propyl Methyl Cellulose (HPMC) during processing of pasta. SDS-PAGE studies indicated changes in High Molecular Weight Glutenin (HMW) fractions during drying stages of pasta preparation and in cooked pasta samples. In uncooked pasta, gel filtration patterns showed four peaks corresponding to different storage proteins whereas in the case of cooked pasta, these peaks were merged into three peaks. Pasta quality characteristics studies indicated that pasta with HPMC was found to have minimum percentage of cooking loss (5.6%), increased cooked weight (82 g), firmness (2.97 N) and high overall quality score (27) than GMS, SSL and control. Microstructure studies confirm the beneficial effect of HPMC. The present study indicated that HPMC is better additive for pasta manufacture followed by GMS. This could be due to interaction of HPMC with starch and protein matrix is different from that of GMS and SSL.

  3. Purification of PRL receptors from toad kidney: Comparisons with rabbit mammary PRL receptors

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Dunand, M.; Kraehenbuhl, J.P.; Rossier, B.C.

    1988-03-01

    The binding characteristics of the prolactin (PRL) receptors present in toad (Bufo marinus) kidneys were investigated and compared to those of PRL receptors present in rabbit mammary glands. The molecular characteristics of the Triton X-100 solubilized renal and mammary PRL receptors were assessed by gel filtration and by migration analysis on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) after affinity labeling of the binding sites with {sup 125}I-human growth hormone. Similar results were obtained for both receptors. Partial purification of the toad PRL receptor could be achieved by affinity chromatography. The molecular weight of this purified receptor could be determined bymore » analysis of SDS-PAGE. With the use of a polyclonal antiserum raised against a purified preparation of rabbit mammary PRL receptor, one or several antigenic epitope(s) could be identified on the core of the toad renal PRL receptor. In conclusion, although the structure and the biological role(s) of PRL have substantially changed during evolution, the receptor for this hormone has retained many of its structural features as could be assessed between an amphibian and a mammalian species on functionally different target tissues.« less

  4. Use of an enzyme-assisted method to improve protein extraction from olive leaves.

    PubMed

    Vergara-Barberán, M; Lerma-García, M J; Herrero-Martínez, J M; Simó-Alfonso, E F

    2015-02-15

    The improvement of protein extraction from olive leaves using an enzyme-assisted protocol has been investigated. Using a cellulase enzyme (Celluclast® 1.5L), different parameters that affect the extraction process, such as the influence and amount of organic solvent, enzyme amount, pH and extraction temperature and time, were optimised. The influence of these factors was examined using the standard Bradford assay and the extracted proteins were characterised by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The optimum extraction parameters were: 30% acetonitrile, 5% (v/v) Celluclast® 1.5L at pH 5.0 and 55°C for 15min. Under these conditions, several protein extracts from olive leaves of different genetic variety (with a total protein amount comprised between 1.87 and 6.64mgg(-1)) were analysed and compared by SDS-PAGE, showing differences in their electrophoretic protein profiles. The developed enzyme-assisted extraction method has shown a faster extraction, higher recovery and reduced solvent usage with respect to the use of the non-enzymatic methods described in literature. Copyright © 2014 Elsevier Ltd. All rights reserved.

  5. Isolation of metallothionein from cells derived from aggressive form of high-grade prostate carcinoma using paramagnetic antibody-modified microbeads off-line coupled with electrochemical and electrophoretic analysis.

    PubMed

    Masarik, Michal; Gumulec, Jaromir; Sztalmachova, Marketa; Hlavna, Marian; Babula, Petr; Krizkova, Sona; Ryvolova, Marketa; Jurajda, Michal; Sochor, Jiri; Adam, Vojtech; Kizek, Rene

    2011-12-01

    Prostate cancer with altered zinc(II) cell metabolism is the second most frequently diagnosed cancer in developed countries. The alterations of zinc(II) metabolism can influence metabolism of other metal ions and can also be associated with the expression and translation of metal-binding proteins including metallothioneins. The aim of this article was to optimize immunoseparation protocol based on paramagnetic beads conjugated with protein G for the isolation of metallothionein. Isolated metallothionein was determined by differential pulse voltammetry Brdicka reaction and SDS-PAGE. Optimal conditions: antigen-binding time - 60 min, temperature - 70°C, and buffer composition and pH - acetate buffer, pH 4.3, were determined. Under the optimized conditions, lysates from 22Rv1 prostate cancer cells treated with various concentrations of cadmium(II) and copper(II) ions were analyzed. We observed strong correlation in all experimental groups and all lysate types (r>0.83 at p<0.041) between metallothionein concentration related to viability and concentration of copper(II) ions and cadmium(II) ions in medium. Moreover, the results were compared with standard sample preparation as heat treatment and SDS-PAGE analysis. Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. Anaphylaxis induced by ingestion of raw garlic.

    PubMed

    Ma, Shikun; Yin, Jia

    2012-08-01

    Patients allergic to garlic often present dermatitis, rhinitis, asthma, and urticaria after ingestion of garlic, contact with garlic, or exposure to garlic dust. Garlic-related anaphylaxis is rare, and the impact of heating on garlic allergens is not very clear. We report a case of anaphylaxis induced by ingestion of raw rather than cooked garlic with manifestations different from previous reports, and we hypothesized that heating could reduce the allergenicity of garlic. Serum total immunoglobulin E (IgE) and specific IgE were tested using the Phadia CAP System FEIA (Phadia, Uppsala, Sweden). Protein extracts from raw and cooked garlic were analyzed by sodium dodecylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. Serum-specific IgE for garlic was 8.16 kUA/L. IgE banding proteins could only be detected in raw garlic extract, because allergens in garlic were mostly degraded into small fragments after heating, as shown in SDS-PAGE profile. In conclusion, raw garlic could induce life-threatening anaphylaxis. However, most of its allergens are heat labile, and patients allergic to garlic might tolerate the cooked one well.

  7. Purification and characterization of nattokinase from Bacillus subtilis natto B-12.

    PubMed

    Wang, Cong; Du, Ming; Zheng, Dongmei; Kong, Fandong; Zu, Guoren; Feng, Yibing

    2009-10-28

    Bacillus subtilis natto B-12 was isolated from natto, a traditional fermented soybean food in Japan. A fibrinolytic enzyme (B-12 nattokinase) was purified from the supernatant of B. subtilis natto B-12 culture broth and showed strong fibrinolytic activity. The enzyme was homogenously purified to 56.1-fold, with a recovery of 43.2% of the initial activity. B-12 nattokinase was demonstrated to be homogeneous by SDS-PAGE and was identified as a monomer of 29000 +/- 300 Da in its native state by SDS-PAGE and size exclusion methods. The optimal pH value and temperature were 8.0 and 40 degrees C, respectively. Purified nattokinase showed high thermostability at temperatures from 30 to 50 degrees C and alkaline stability within the range of pH 6.0-9.0. The enzyme activity was activated by Zn(2+) and obviously inhibited by Fe(3+) and Al(3+). This study provides some important information for the effect factors of fibrinolytic activity, the purification methods, and characterization of nattokinase from B. subtilis natto B-12, which enriches the theoretical information of nattokinase for the research and development of nattokinase as a functional additive of food.

  8. Aphid Gel Saliva: Sheath Structure, Protein Composition and Secretory Dependence on Stylet-Tip Milieu

    PubMed Central

    Will, Torsten; Steckbauer, Kathrin; Hardt, Martin; van Bel, Aart J. E.

    2012-01-01

    In order to separate and analyze saliva types secreted during stylet propagation and feeding, aphids were fed on artificial diets. Gel saliva was deposited as chains of droplets onto Parafilm membranes covering the diets into which watery saliva was secreted. Saliva compounds collected from the diet fluid were separated by SDS-PAGE, while non-soluble gel saliva deposits were processed in a novel manner prior to protein separation by SDS-PAGE. Soluble (watery saliva) and non-soluble (gel saliva) protein fractions were significantly different. To test the effect of the stylet milieu on saliva secretion, aphids were fed on various diets. Hardening of gel saliva is strongly oxygen-dependent, probably owing to formation of sulfide bridges by oxidation of sulphydryl groups. Surface texture of gel saliva deposits is less pronounced under low-oxygen conditions and disappears in dithiothreitol containing diet. Using diets mimicking sieve-element sap and cell-wall fluid respectively showed that the soluble protein fraction was almost exclusively secreted in sieve elements while non-soluble fraction was preferentially secreted at cell wall conditions. This indicates that aphids are able to adapt salivary secretion in dependence of the stylet milieu. PMID:23056521

  9. Identification of Tropomyosin and Its Immunological Properties from Larvae of Cattle Tick, Boophilus annulatus

    PubMed Central

    Nabian, S; Taheri, M; Fard, R Mazaheri Nezhad; Aramoon, M

    2013-01-01

    Background Boophilus annulatus is an obligate blood feeder tick that can cause great losses in animals due to anemia and its ability to injure its host skin directly. The aim of this study was identification of cattle humoral immune response to some tick proteins during experimental infestation. Methods Immune sera against tick were collected from experimentally infested cattle with ticks. One and two-dimensional electrophoresis and Western blotting methods were used for the detection of immunogenic proteins in larval tick extract and eight of these proteins were identified by MALDITOF and MALDI-TOF-TOF mass spectrometry. Results In non-reducing one-dimensional SDS-PAGE, some bounds between 12 to more than 250-kDa appeared. In two-dimensional SDS-PAGE, numerous spot appeared and the identified immunogenic proteins by parallel immunoblotting weighted between 14 and 97 kDa. Amino acid sequences of protein spot with 37-kDa molecular weight had identity to tropomyosin based on Mascot search in NCBI. Conclusion Anti tropomyosin antibodies can be induced in experimentally infested hosts with ticks and it seems that tropomyosin can be useful for the development of anti tick vaccines. PMID:23914237

  10. Auxin effects on in vitro and in vivo protein phosphorylation in pea. [Pisum sativum

    DOE Office of Scientific and Technical Information (OSTI.GOV)

    Gallagher, S.R.; Ray, P.M.

    1987-04-01

    Terminal 8mm sections from the third internode of dark grown 7 day old Pisum sativum cv Alaska seedlings were separated into membrane and soluble fractions. SDS gradient PAGE identified approximately 50 in vivo phosphorylated proteins and proved superior to 2-D SDS PAGE in terms of resolution and repeatability. Addition of indoleacetic acid (IAA), fusicoccin, or 2,4 dichlorophenoxyacetic acid to membranes resulted in no detectable change in the number or phosphorylation level of the labeled proteins during in vitro phosphorylation in the presence of submicromolar concentrations of calcium. Similar results were obtained with soluble proteins. In the absence of calcium, themore » level of in vitro protein phosphorylation was much less, but not auxin effects could be identified. Furthermore, treatment of the sections with IAA in vivo followed by cell fractionation and in vitro phosphorylation failed to identify auxin responsive proteins. Lastly, when sections were labeled with /sup 32/P inorganic phosphate in the presence of 17 uM IAA, no auxin specific changes were found in the level of phosphorylation or in the number of phosphorylated proteins. Auxin effects on phosphorylation are thus slight or below their detection limit.« less

  11. Purification and characterization of thermostable alpha-galactosidase from Aspergillus terreus (GR).

    PubMed

    Shankar, S K; Dhananjay, S K; Mulimani, V H

    2009-02-01

    An extracellular thermostable alpha-galactosidase producing Aspergillus terreus (GR) strain was isolated from soil sample using guar gum as sole source of carbon. It was purified to apparent homogeneity by acetone precipitation, gel filtration followed by DEAE-Sephacel chromatographic step. The purified enzyme showed a single band after sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight of the purified enzyme after SDS-PAGE was 108 kDa. The enzyme showed optimum pH and temperature of 5.0 and 65 degrees C, respectively, for artificial substrate pNPalphaGal. alpha-Galactosidase from A. terreus (GR) is found to be thermostable, as it was not inactivated after heating at 65 degrees C for 40 min. The K (m) for pNPalphaGal, oNPalphaGal, raffinose, and stachyose are 0.1, 0.28, 0.42, and 0.33 mM, respectively. Inhibitors such as 1,10-phenanthroline, phenylmethylsulfonyl fluoride, ethylenediaminetetraacetic acid, mercaptoethanol, and urea have no effect, whereas N-bromosuccinamide inhibited enzyme activity by 100%. Among metal ions tested, Mg(2+), Ni(2+), Ca(2+), Co(2+), and Mn(2+) had no effect on enzyme activity, but Ag(+), Hg(2+), and Cu(2+) have inhibited complete activity.

  12. Identification of Yeast V-ATPase Mutants by Western Blots Analysis of Whole Cell Lysates

    NASA Astrophysics Data System (ADS)

    Parra-Belky, Karlett

    2002-11-01

    A biochemistry laboratory was designed for an undergraduate course to help students better understand the link between molecular engineering and biochemistry. Students identified unknown yeast strains with high specificity using SDS-PAGE and Western blot analysis of whole cell lysates. This problem-solving exercise is a common application of biochemistry in biotechnology research. Three different strains were used: a wild-type and two mutants for the proton pump vacuolar ATPase (V-ATPase). V-ATPases are multisubunit enzymes and the mutants used were deletion mutants; each lacked one structural gene of the complex. After three, three-hour labs, mutant strains were easily identified by the students and distinguished from wild-type cells analyzing the pattern of SDS-PAGE distribution of proteins. Identifying different subunits of one multimeric protein allowed for discussion of the structure and function of this metabolic enzyme, which captured the interest of the students. The experiment can be adapted to other multimeric protein complexes and shows improvement of the described methodology over previous reports, perhaps because the problem and its solution are representative of the type of techniques currently used in research labs.

  13. Evaluating nodes importance in complex network based on PageRank algorithm

    NASA Astrophysics Data System (ADS)

    Li, Kai; He, Yongfeng

    2018-04-01

    To evaluate the important nodes in the complex network, and aim at the problems existing in the traditional PageRank algorithm, we propose a modified PageRank algorithm. The algorithm has convergence for the evaluation of the importance of the suspended nodes and the nodes with a directed loop network. The simulation example shows the effectiveness of the modified algorithm for the evaluation of the complexity of the complex network nodes.

  14. Building Interactive Simulations in Web Pages without Programming.

    PubMed

    Mailen Kootsey, J; McAuley, Grant; Bernal, Julie

    2005-01-01

    A software system is described for building interactive simulations and other numerical calculations in Web pages. The system is based on a new Java-based software architecture named NumberLinX (NLX) that isolates each function required to build the simulation so that a library of reusable objects could be assembled. The NLX objects are integrated into a commercial Web design program for coding-free page construction. The model description is entered through a wizard-like utility program that also functions as a model editor. The complete system permits very rapid construction of interactive simulations without coding. A wide range of applications are possible with the system beyond interactive calculations, including remote data collection and processing and collaboration over a network.

  15. 47 CFR 95.27 - Paging receiver description.

    Code of Federal Regulations, 2010 CFR

    2010-10-01

    ... receiver is a unit capable of receiving the radio signals from a base station for the bearer to hear a page (someone's name or other identifier said in order to find, summon or notify him/her) spoken by the base station operator. ...

  16. 7 CFR 3402.11 - Proposal cover page.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 Agriculture 15 2010-01-01 2010-01-01 false Proposal cover page. 3402.11 Section 3402.11 Agriculture Regulations of the Department of Agriculture (Continued) COOPERATIVE STATE RESEARCH, EDUCATION, AND EXTENSION SERVICE, DEPARTMENT OF AGRICULTURE FOOD AND AGRICULTURAL SCIENCES NATIONAL NEEDS...

  17. Non-Gradient Blue Native Polyacrylamide Gel Electrophoresis.

    PubMed

    Luo, Xiaoting; Wu, Jinzi; Jin, Zhen; Yan, Liang-Jun

    2017-02-02

    Gradient blue native polyacrylamide gel electrophoresis (BN-PAGE) is a well established and widely used technique for activity analysis of high-molecular-weight proteins, protein complexes, and protein-protein interactions. Since its inception in the early 1990s, a variety of minor modifications have been made to this gradient gel analytical method. Here we provide a major modification of the method, which we call non-gradient BN-PAGE. The procedure, similar to that of non-gradient SDS-PAGE, is simple because there is no expensive gradient maker involved. The non-gradient BN-PAGE protocols presented herein provide guidelines on the analysis of mitochondrial protein complexes, in particular, dihydrolipoamide dehydrogenase (DLDH) and those in the electron transport chain. Protocols for the analysis of blood esterases or mitochondrial esterases are also presented. The non-gradient BN-PAGE method may be tailored for analysis of specific proteins according to their molecular weight regardless of whether the target proteins are hydrophobic or hydrophilic. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

  18. Readability and quality of wikipedia pages on neurosurgical topics.

    PubMed

    Modiri, Omeed; Guha, Daipayan; Alotaibi, Naif M; Ibrahim, George M; Lipsman, Nir; Fallah, Aria

    2018-03-01

    Wikipedia is the largest online encyclopedia with over 40 million articles, and generating 500 million visits per month. The aim of this study is to assess the readability and quality of Wikipedia pages on neurosurgical related topics. We selected the neurosurgical related Wikipedia pages based on the series of online patient information articles that are published by the American Association of Neurological Surgeons (AANS). We assessed readability of Wikipedia pages using five different readability scales (Flesch Reading Ease, Flesch Kincaid Grade Level, Gunning Fog Index, SMOG) Grade level, and Coleman-Liau Index). We used the Center for Disease Control (CDC) Clear Communication Index as well as the DISCERN Instrument to evaluate the quality of each Wikipedia article. We identified a total of fifty-five Wikipedia articles that corresponded with patient information articles published by the AANS. This constitutes 77.46% of the AANS topics. The mean Flesch Kincaid reading ease score for all of the Wikipedia articles we analyzed is 31.10, which indicates that a college-level education is necessary to understand them. In comparison to the readability analysis for the AANS articles, the Wikipedia articles were more difficult to read across every scale. None of the Wikipedia articles meet the CDC criterion for clear communications. Our analyses demonstrated that Wikipedia articles related to neurosurgical topics are associated with higher grade levels for reading and also below the expected levels of clear communications for patients. Collaborative efforts from the neurosurgical community are needed to enhance the readability and quality of Wikipedia pages related to neurosurgery. Copyright © 2018 Elsevier B.V. All rights reserved.

  19. Planetary Photojournal Home Page Graphic

    NASA Technical Reports Server (NTRS)

    2004-01-01

    This image is an unannotated version of the Planetary Photojournal Home Page graphic. This digital collage contains a highly stylized rendition of our solar system and points beyond. As this graphic was intended to be used as a navigation aid in searching for data within the Photojournal, certain artistic embellishments have been added (color, location, etc.). Several data sets from various planetary and astronomy missions were combined to create this image.

  20. Photojournal Home Page Graphic 2007

    NASA Technical Reports Server (NTRS)

    2008-01-01

    This image is an unannotated version of the Photojournal Home Page graphic released in October 2007. This digital collage contains a highly stylized rendition of our solar system and points beyond. As this graphic was intended to be used as a navigation aid in searching for data within the Photojournal, certain artistic embellishments have been added (color, location, etc.). Several data sets from various planetary and astronomy missions were combined to create this image.