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Sample records for addition northern blot

  1. BlotBase: a northern blot database.

    PubMed

    Schlamp, K; Weinmann, A; Krupp, M; Maass, T; Galle, Pr; Teufel, A

    2008-12-31

    With the availability of high-throughput gene expression analysis, multiple public expression databases emerged, mostly based on microarray expression data. Although these databases are of significant biomedical value, they do hold significant drawbacks, especially concerning the reliability of single gene expression profiles obtained by microarray data. Simultaneously, reliable data on an individual gene's expression are often published as single northern blots in individual publications. These data were not yet available for high-throughput screening. To reduce the gap between high-throughput expression data and individual highly reliable expression data, we designed a novel database "BlotBase", a freely and easily accessible database, currently containing approximately 700 published northern blots of human or mouse origin (http://www.medicalgenomics.org/Databases/BlotBase). As the database is open for public data submission, we expect this database to quickly become a large expression profiling resource, eventually providing higher reliability in high-throughput gene expression analysis. Realizing BlotBase, Pubmed was searched manually and by computer based text mining methods to obtain publications containing northern blot results. Subsequently, northern blots were extracted and expression values of different tissues calculated utilizing Image J. All data were made available through a user friendly web front end. The data may be searched by either full text search or list of available northern blots of a specific tissue. Northern blot expression profiles were displayed by three expression states as well as a bar chart, allowing for automated evaluation. Furthermore, we integrated additional features, e.g. instant access to the corresponding RNA sequence or primer design tools making further expression analysis more convenient. Finally, through a semiautomatic submission system this database was opened to the bioinformatics community. PMID:18838116

  2. Multiplexed miRNA northern blots via hybridization chain reaction.

    PubMed

    Schwarzkopf, Maayan; Pierce, Niles A

    2016-09-01

    Northern blots enable detection of a target RNA of interest in a biological sample using standard benchtop equipment. miRNAs are the most challenging targets as they must be detected with a single short nucleic acid probe. With existing approaches, it is cumbersome to perform multiplexed blots in which several RNAs are detected simultaneously, impeding the study of interacting regulatory elements. Here, we address this shortcoming by demonstrating multiplexed northern blotting based on the mechanism of hybridization chain reaction (HCR). With this approach, nucleic acid probes complementary to RNA targets trigger chain reactions in which fluorophore-labeled DNA hairpins self-assemble into tethered fluorescent amplification polymers. The programmability of HCR allows multiple amplifiers to operate simultaneously and independently within a blot, enabling straightforward multiplexing. We demonstrate simultaneous detection of three endogenous miRNAs in total RNA extracted from 293T and HeLa cells. For a given target, HCR signal scales linearly with target abundance, enabling relative and absolute quantitation. Using non-radioactive HCR, sensitive and selective miRNA detection is achieved using 2'OMe-RNA probes. The HCR northern blot protocol takes ∼1.5 days independent of the number of target RNAs. PMID:27270083

  3. Multiplexed miRNA northern blots via hybridization chain reaction

    PubMed Central

    Schwarzkopf, Maayan; Pierce, Niles A.

    2016-01-01

    Northern blots enable detection of a target RNA of interest in a biological sample using standard benchtop equipment. miRNAs are the most challenging targets as they must be detected with a single short nucleic acid probe. With existing approaches, it is cumbersome to perform multiplexed blots in which several RNAs are detected simultaneously, impeding the study of interacting regulatory elements. Here, we address this shortcoming by demonstrating multiplexed northern blotting based on the mechanism of hybridization chain reaction (HCR). With this approach, nucleic acid probes complementary to RNA targets trigger chain reactions in which fluorophore-labeled DNA hairpins self-assemble into tethered fluorescent amplification polymers. The programmability of HCR allows multiple amplifiers to operate simultaneously and independently within a blot, enabling straightforward multiplexing. We demonstrate simultaneous detection of three endogenous miRNAs in total RNA extracted from 293T and HeLa cells. For a given target, HCR signal scales linearly with target abundance, enabling relative and absolute quantitation. Using non-radioactive HCR, sensitive and selective miRNA detection is achieved using 2′OMe-RNA probes. The HCR northern blot protocol takes ∼1.5 days independent of the number of target RNAs. PMID:27270083

  4. Quantitative Northern Blot Analysis of Mammalian rRNA Processing.

    PubMed

    Wang, Minshi; Pestov, Dimitri G

    2016-01-01

    Assembly of eukaryotic ribosomes is an elaborate biosynthetic process that begins in the nucleolus and requires hundreds of cellular factors. Analysis of rRNA processing has been instrumental for studying the mechanisms of ribosome biogenesis and effects of stress conditions on the molecular milieu of the nucleolus. Here, we describe the quantitative analysis of the steady-state levels of rRNA precursors, applicable to studies in mammalian cells and other organisms. We include protocols for gel electrophoresis and northern blotting of rRNA precursors using procedures optimized for the large size of these RNAs. We also describe the ratio analysis of multiple precursors, a technique that facilitates the accurate assessment of changes in the efficiency of individual pre-rRNA processing steps. PMID:27576717

  5. Optimization of northern analysis by vacuum-blotting, RNA-transfer visualization, and ultraviolet fixation

    SciTech Connect

    Kroczek, R.A.; Siebert, E. )

    1990-01-01

    We have optimized Northern analysis at several steps. Overnight electrophoresis was replaced by short gel runs and overnight capillary transfer by rapid vacuum-blotting adapted to Northern analysis. Short uv irradiation was used as a substitute for the usual RNA fixation by baking. Direct staining of RNA before electrophoresis made it possible to check RNA integrity and to evaluate the quality of the size separation immediately after electrophoresis. In this system, RNA transfer onto the membrane support could also be quickly assessed after the blotting step. The net result of all modifications was a doubling of the autoradiography signal compared with that obtained by modern Northern protocols. At the same time, the duration of the procedure was shortened drastically, allowing an autoradiography signal to be obtained within 24 h.

  6. Immuno-Northern Blotting: Detection of RNA Modifications by Using Antibodies against Modified Nucleosides.

    PubMed

    Mishima, Eikan; Jinno, Daisuke; Akiyama, Yasutoshi; Itoh, Kunihiko; Nankumo, Shinnosuke; Shima, Hisato; Kikuchi, Koichi; Takeuchi, Yoichi; Elkordy, Alaa; Suzuki, Takehiro; Niizuma, Kuniyasu; Ito, Sadayoshi; Tomioka, Yoshihisa; Abe, Takaaki

    2015-01-01

    The biological roles of RNA modifications are still largely not understood. Thus, developing a method for detecting RNA modifications is important for further clarification. We developed a method for detecting RNA modifications called immuno-northern blotting (INB) analysis and herein introduce its various capabilities. This method involves the separation of RNAs using either polyacrylamide or agarose gel electrophoresis, followed by transfer onto a nylon membrane and subsequent immunoblotting using antibodies against modified nucleosides for the detection of specific modifications. We confirmed that INB with the antibodies for 1-methyladenosine (m1A), N6-methyladenosine (m6A), pseudouridine, and 5-methylcytidine (m5C) showed different modifications in a variety of RNAs from various species and organelles. INB with the anti-m5C antibody revealed that the antibody cross-reacted with another modification on DNA, suggesting the application of this method for characterization of the antibody for modified nucleosides. Additionally, using INB with the antibody for m1A, which is a highly specific modification in eukaryotic tRNA, we detected tRNA-derived fragments known as tiRNAs under the cellular stress response, suggesting the application for tracking target RNA containing specific modifications. INB with the anti-m6A antibody confirmed the demethylation of m6A by the specific demethylases fat mass and obesity-associated protein (FTO) and ALKBH5, suggesting its application for quantifying target modifications in separated RNAs. Furthermore, INB demonstrated that the knockdown of FTO and ALKBH5 increased the m6A modification in small RNAs as well as in mRNA. The INB method has high specificity, sensitivity, and quantitative capability, and it can be employed with conventional experimental apparatus. Therefore, this method would be useful for research on RNA modifications and metabolism. PMID:26606401

  7. Immuno-Northern Blotting: Detection of RNA Modifications by Using Antibodies against Modified Nucleosides

    PubMed Central

    Akiyama, Yasutoshi; Itoh, Kunihiko; Nankumo, Shinnosuke; Shima, Hisato; Kikuchi, Koichi; Takeuchi, Yoichi; Elkordy, Alaa; Suzuki, Takehiro; Niizuma, Kuniyasu; Ito, Sadayoshi; Tomioka, Yoshihisa; Abe, Takaaki

    2015-01-01

    The biological roles of RNA modifications are still largely not understood. Thus, developing a method for detecting RNA modifications is important for further clarification. We developed a method for detecting RNA modifications called immuno-northern blotting (INB) analysis and herein introduce its various capabilities. This method involves the separation of RNAs using either polyacrylamide or agarose gel electrophoresis, followed by transfer onto a nylon membrane and subsequent immunoblotting using antibodies against modified nucleosides for the detection of specific modifications. We confirmed that INB with the antibodies for 1-methyladenosine (m1A), N6-methyladenosine (m6A), pseudouridine, and 5-methylcytidine (m5C) showed different modifications in a variety of RNAs from various species and organelles. INB with the anti-m5C antibody revealed that the antibody cross-reacted with another modification on DNA, suggesting the application of this method for characterization of the antibody for modified nucleosides. Additionally, using INB with the antibody for m1A, which is a highly specific modification in eukaryotic tRNA, we detected tRNA-derived fragments known as tiRNAs under the cellular stress response, suggesting the application for tracking target RNA containing specific modifications. INB with the anti-m6A antibody confirmed the demethylation of m6A by the specific demethylases fat mass and obesity-associated protein (FTO) and ALKBH5, suggesting its application for quantifying target modifications in separated RNAs. Furthermore, INB demonstrated that the knockdown of FTO and ALKBH5 increased the m6A modification in small RNAs as well as in mRNA. The INB method has high specificity, sensitivity, and quantitative capability, and it can be employed with conventional experimental apparatus. Therefore, this method would be useful for research on RNA modifications and metabolism. PMID:26606401

  8. Exploring the Foundation of Genomics: A Northern Blot Reference set for the Comparative Analysis of Transcript Profiling Technologies

    PubMed Central

    Kemmer, Danielle; Faxén, Margareta; Hodges, Emily; Lim, Jonathan; Herzog, Elena; Ljungström, Elsebrit; Lundmark, Anders; Olsen, Mary K.; Podowski, Raf; Sonnhammer, Erik L. L.; Nilsson, Peter; Reimers, Mark; Lenhard, Boris; Roberds, Steven L.; Wahlestedt, Claes; Höög, Christer; Agarwal, Pankaj

    2004-01-01

    In this paper we aim to create a reference data collection of Northern blot results and demonstrate how such a collection can enable a quantitative comparison of modern expression profiling techniques, a central component of functional genomics studies. Historically, Northern blots were the de facto standard for determining RNA transcript levels. However, driven by the demand for analysis of large sets of genes in parallel, high-throughput methods, such as microarrays, dominate modern profiling efforts. To facilitate assessment of these methods, in comparison to Northern blots, we created a database of published Northern results obtained with a standardized commercial multiple tissue blot (dbMTN). In order to demonstrate the utility of the dbMTN collection for technology comparison, we also generated expression profiles for genes across a set of human tissues, using multiple profiling techniques. No method produced profiles that were strongly correlated with the Northern blot data. The highest correlations to the Northern blot data were determined with microarrays for the subset of genes observed to be specifically expressed in a single tissue in the Northern analyses. The database and expression profiling data are available via the project website (http://www.cisreg.ca). We believe that emphasis on multitechnique validation of expression profiles is justified, as the correlation results between platforms are not encouraging on the whole. Supplementary material for this article can be found at: http://www.interscience.wiley.com/jpages/1531-6912/suppmat PMID:18629180

  9. Comparison of the rotavirus nonstructural protein NSP1 (NS53) from different species by sequence analysis and northern blot hybridization.

    PubMed

    Dunn, S J; Cross, T L; Greenberg, H B

    1994-08-15

    The nucleotide sequence of gene 5 encoding the rotavirus nonstructural protein NSP1 (NS53) of 6 strains (EW, EHP, RRV, I321, OSU, and Gottfried) was determined and compared to 6 previously reported strains (SA11, UK, RF, Hu803, DS-1, and Wa). The 12 rotavirus strains were derived from a total of five separate species (murine, bovine, simian, porcine, and human). Gene sizes ranged from 1564 to 1611 nucleotides in length and the deduced protein sequences were found to be 486 to 495 amino acids in length. Comparisons of NSP1 amino acid sequences showed identities ranging from 36 to 92%. This diversity was most evident between strains from different species. Phylogenetic analysis revealed a clustering of NSP1 sequences according to species origin with the exception that the human and porcine strains were included in a single grouping. Northern blot hybridizations using additional rotavirus strains from the five species confirmed the grouping found by sequence analysis. The species specificity of NSP1 is consistent with the hypothesis that NSP1 plays a role in host range restriction. PMID:8030275

  10. The gene encoding vitamin K-dependent anticoagulant protein S is expressed in multiple rabbit organs as demonstrated by northern blotting, in situ hybridization, and immunohistochemistry.

    PubMed

    He, X; Shen, L; Bjartell, A; Dahlbäck, B

    1995-01-01

    Vitamin K-dependent protein S is an anticoagulant plasma protein that functions as a co-factor to activated protein C in the degradation of coagulation factors Va and VIIIa. We investigated the tissue/cellular distribution of protein S synthesis by Northern blotting, in situ hybridization, and immunohistochemistry. Northern blotting together with in situ hybridization, using specific oligodeoxynucleotide probes, demonstrated protein S mRNA in liver, lung, testis, epididymis, ovary, uterus, and brain. In the reproductive system, protein S mRNA was present in the cytoplasm of Leydig cells, interstitial cells of the ovary, epithelial cells of the epididymis, and in the endometrium, including endometrial mucous glandular membrane in the myometrium. Bronchial epithelial cells and alveolar macrophages were positive in the respiratory system. In the central nervous system, pyramidal neurons in the cerebral cortex and in the hippocampal region, and dentate fascia neurons gave strongly positive signals. Immunohistochemistry with monoclonal antibodies yielded a staining pattern that correlated well with results of in situ hybridization. In conclusion, results from Northern blotting, in situ hybridization, and immunohistochemistry suggested that rabbit protein S is expressed in several extrahepatic tissues. The presence of protein S transcripts in these fully differentiated cells suggests a cell type-specific gene expression which may be related to local anticoagulation or to other as yet unknown protein S functions. PMID:7822769

  11. Southern blotting.

    PubMed

    Brown, T

    2001-05-01

    Southern blotting is the transfer of DNA fragments from an electrophoresis gel to a membrane support (the properties and advantages of the different types of membrane, transfer buffer, and transfer method are discussed in detail), resulting in immobilization of the DNA fragments, so the membrane carries a semipermanent reproduction of the banding pattern of the gel. After immobilization, the DNA can be subjected to hybridization analysis, enabling bands with sequence similarity to a labeled probe to be identified. This appendix describes Southern blotting via upward capillary transfer of DNA from an agarose gel onto a nylon or nitrocellulose membrane, using a high-salt transfer buffer to promote binding of DNA to the membrane. With the high-salt buffer, the DNA becomes bound to the membrane during transfer but not permanently immobilized. Immobilization is achieved by UV irradiation (for nylon) or baking (for nitrocellulose). A Support Protocol describes how to calibrate a UV transilluminator for optimal UV irradiation of a nylon membrane. An alternate protocol details transfer using nylon membranes and an alkaline buffer, and is primarily used with positively charged nylon membranes. The advantage of this combination is that no post-transfer immobilization step is required, as the positively charged membrane binds DNA irreversibly under alkaline transfer conditions. The method can also be used with neutral nylon membranes but less DNA will be retained. A second alternate protocol describes a transfer method based on a different transfer-stack setup. The traditional method of upward capillary transfer of DNA from gel to membrane described in the first basic and alternate protocols has certain disadvantages, notably the fact that the gel can become crushed by the weighted filter papers and paper towels that are laid on top of it. This slows down the blotting process and may reduce the amount of DNA that can be transferred. The downward capillary method described in

  12. Southwestern Blotting Assay

    PubMed Central

    Jia, Yinshan; Nagore, Linda; Jarrett, Harry

    2016-01-01

    Southwestern blotting is a technique used to study DNA-protein interactions. This method detects specific DNA-binding proteins by incubating radiolabeled DNA with a gel blot, washing, and visualizing through autoradiography. A blot resulting from 1-dimensional SDS-PAGE reveals the molecular weight of the binding proteins. To increase separation and determine isoelectric point a 2-dimensional gel can be blotted. Additional dimensions of electrophoresis, such as a gel shift (EMSA), can precede isoelectric focusing and SDS-PAGE to further improve separation. Combined with other techniques, such as mass spectrometry, the DNA-binding protein can be identified. PMID:26404144

  13. VIEW OF CEMETERY SECTION NA (NEW ADDITION), WITH NORTHERN PERIMETER ...

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

    VIEW OF CEMETERY SECTION NA (NEW ADDITION), WITH NORTHERN PERIMETER FENCE ALONG LINCOLN BOULEVARD IN FOREGROUND. VIEW TO SOUTH. - San Francisco National Cemetery, 1 Lincoln Boulevard, San Francisco, San Francisco County, CA

  14. MicroRNA fate upon targeting with anti-miRNA oligonucleotides as revealed by an improved Northern-blot-based method for miRNA detection

    PubMed Central

    Torres, Adrian G.; Fabani, Martin M.; Vigorito, Elena; Gait, Michael J.

    2011-01-01

    MicroRNAs (miRNAs) are small non-coding RNAs involved in fine-tuning of gene regulation. Antisense oligonucleotides (ONs) are promising tools as anti-miRNA (anti-miR) agents toward therapeutic applications and to uncover miRNA function. Such anti-miR ONs include 2′-O-methyl (OMe), cationic peptide nucleic acids like K-PNA-K3, and locked nucleic acid (LNA)-based anti-miRs such as LNA/DNA or LNA/OMe. Northern blotting is a widely used and robust technique to detect miRNAs. However, miRNA quantification in the presence of anti-miR ONs has proved to be challenging, due to detection artifacts, which has led to poor understanding of miRNA fate upon anti-miR binding. Here we show that anti-miR ON bound to miR-122 can prevent the miRNA from being properly precipitated into the purified RNA fraction using the standard RNA extraction protocol (TRI-Reagent), yielding an RNA extract that does not reflect the real cellular levels of the miRNA. An increase in the numbers of equivalents of isopropanol during the precipitation step leads to full recovery of the targeted miRNA back into the purified RNA extract. Following our improved protocol, we demonstrate by Northern blotting, in conjunction with a PNA decoy strategy and use of high denaturing PAGE, that high-affinity anti-miRs (K-PNA-K3, LNA/DNA, and LNA/OMe) sequester miR-122 without causing miRNA degradation, while miR-122 targeting with a lower-affinity anti-miR (OMe) seems to promote degradation of the miRNA. The technical issues explored in this work will have relevance for other hybridization-based techniques for miRNA quantification in the presence of anti-miR ONs. PMID:21441346

  15. MicroRNA fate upon targeting with anti-miRNA oligonucleotides as revealed by an improved Northern-blot-based method for miRNA detection.

    PubMed

    Torres, Adrian G; Fabani, Martin M; Vigorito, Elena; Gait, Michael J

    2011-05-01

    MicroRNAs (miRNAs) are small non-coding RNAs involved in fine-tuning of gene regulation. Antisense oligonucleotides (ONs) are promising tools as anti-miRNA (anti-miR) agents toward therapeutic applications and to uncover miRNA function. Such anti-miR ONs include 2'-O-methyl (OMe), cationic peptide nucleic acids like K-PNA-K3, and locked nucleic acid (LNA)-based anti-miRs such as LNA/DNA or LNA/OMe. Northern blotting is a widely used and robust technique to detect miRNAs. However, miRNA quantification in the presence of anti-miR ONs has proved to be challenging, due to detection artifacts, which has led to poor understanding of miRNA fate upon anti-miR binding. Here we show that anti-miR ON bound to miR-122 can prevent the miRNA from being properly precipitated into the purified RNA fraction using the standard RNA extraction protocol (TRI-Reagent), yielding an RNA extract that does not reflect the real cellular levels of the miRNA. An increase in the numbers of equivalents of isopropanol during the precipitation step leads to full recovery of the targeted miRNA back into the purified RNA extract. Following our improved protocol, we demonstrate by Northern blotting, in conjunction with a PNA decoy strategy and use of high denaturing PAGE, that high-affinity anti-miRs (K-PNA-K3, LNA/DNA, and LNA/OMe) sequester miR-122 without causing miRNA degradation, while miR-122 targeting with a lower-affinity anti-miR (OMe) seems to promote degradation of the miRNA. The technical issues explored in this work will have relevance for other hybridization-based techniques for miRNA quantification in the presence of anti-miR ONs. PMID:21441346

  16. The western blot

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Western blotting is a technique that involves the separation of proteins by gel electrophoresis, their blotting or transfer to a membrane, and selective immunodetection of an immobilized antigen. This is an important and routine method for protein analysis that depends on the specificity of antibod...

  17. The Dot Blot ELISA.

    ERIC Educational Resources Information Center

    Gerbig, Donald G., Jr.; Fenk, Christopher J.; Goodhart, Amy S.

    2000-01-01

    Uses two laboratory techniques, Enzyme Linked Immunosorbent Assay (ELISA) and Western Blot, to demonstrate antibody-antigen binding concepts. Includes a list of required materials and directions for the procedure, and makes suggestions for classroom applications. (Contains 13 references.) (YDS)

  18. Genomic Southern blot analysis.

    PubMed

    Gebbie, Leigh

    2014-01-01

    This chapter describes a detailed protocol for genomic Southern blot analysis which can be used to detect transgene or endogenous gene sequences in cereal genomes. The protocol follows a standard approach that has been shown to generate high-quality results: size fractionation of genomic DNA; capillary transfer to a nylon membrane; hybridization with a digoxigenin-labelled probe; and detection using a chemiluminescent-based system. High sensitivity and limited background are key to successful Southern blots. The critical steps in this protocol are complete digestion of the right quantity of DNA, careful handling of the membrane to avoid unnecessary background, and optimization of probe concentration and temperatures during the hybridization step. Detailed instructions on how to successfully master these techniques are provided. PMID:24243203

  19. Single-cell western blotting

    PubMed Central

    Hughes, Alex J.; Spelke, Dawn P.; Xu, Zhuchen; Kang, Chi-Chih; Schaffer, David V.; Herr, Amy E.

    2014-01-01

    To measure cell-to-cell variation in protein-mediated functions — a hallmark of biological processes — we developed an approach to conduct ~103 concurrent single-cell western blots (scWesterns) in ~4 hours. A microscope slide supporting a 30 µm-thick photoactive polyacrylamide gel enables western blotting comprised of: settling of single cells into microwells, lysis in situ, gel electrophoresis, photoinitiated blotting to immobilize proteins, and antibody probing. We apply this scWestern to monitor single rat neural stem cell differentiation and responses to mitogen stimulation. The scWestern quantifies target proteins even with off-target antibody binding, multiplexes to 11 protein targets per single cell with detection thresholds of <30,000 molecules, and supports analyses of low starting cell numbers (~200) when integrated with fluorescence activated cell sorting. The scWestern thus overcomes limitations in single-cell protein analysis (i.e., antibody fidelity, sensitivity, and starting cell number) and constitutes a versatile tool for the study of complex cell populations at single-cell resolution. PMID:24880876

  20. BLOT Ver. 1.65

    2009-03-24

    BLOT is a graphic program for post-processing finite element analyses output in the EXODUS II database format. It is command driven with free-format input and can drive graphics devices supported by the Sandia Virtual Device Interface. BLOT produces mesh plots of the analysis output variables including deformed mesh plots, line contours, filled (painted) contours, vector plots of two/three variables (velocity vectors), and symbol plots of scalar variables (discrete cracks). Features include pathlines of analysis variablesmore » drawn on the mesh, element selection by material, element birth and death, multiple views combining several displays on each plot, symmetry mirroring, and node and element numbering. X-Y plots of the analysis variables include time vs. variable plots or variable vs. variable plots, and distance vs. variable plots at selected time steps where distance is the accumulated distance between pairs of nodes or element centers. BLOT is written in as portable a form as possible. Fortran code is written in ANSI Standard FORTRAN-77. Machine-specific routines are limited in number and are grouped together to minimize the time required to adapt them to a new system. SEACAS codes have been ported to several Unix systems« less

  1. BLOT Ver. 1.65

    SciTech Connect

    MEYERS, RAY; GLICK, III, JOHN; FORSYTHE, CHRISTI; GILKEY, AMY; SJAARDEMA, GREGORY

    2009-03-24

    BLOT is a graphic program for post-processing finite element analyses output in the EXODUS II database format. It is command driven with free-format input and can drive graphics devices supported by the Sandia Virtual Device Interface. BLOT produces mesh plots of the analysis output variables including deformed mesh plots, line contours, filled (painted) contours, vector plots of two/three variables (velocity vectors), and symbol plots of scalar variables (discrete cracks). Features include pathlines of analysis variables drawn on the mesh, element selection by material, element birth and death, multiple views combining several displays on each plot, symmetry mirroring, and node and element numbering. X-Y plots of the analysis variables include time vs. variable plots or variable vs. variable plots, and distance vs. variable plots at selected time steps where distance is the accumulated distance between pairs of nodes or element centers. BLOT is written in as portable a form as possible. Fortran code is written in ANSI Standard FORTRAN-77. Machine-specific routines are limited in number and are grouped together to minimize the time required to adapt them to a new system. SEACAS codes have been ported to several Unix systems

  2. Multiplexed Western Blotting Using Microchip Electrophoresis.

    PubMed

    Jin, Shi; Furtaw, Michael D; Chen, Huaxian; Lamb, Don T; Ferguson, Stephen A; Arvin, Natalie E; Dawod, Mohamed; Kennedy, Robert T

    2016-07-01

    Western blotting is a commonly used protein assay that combines the selectivity of electrophoretic separation and immunoassay. The technique is limited by long time, manual operation with mediocre reproducibility, and large sample consumption, typically 10-20 μg per assay. Western blots are also usually used to measure only one protein per assay with an additional housekeeping protein for normalization. Measurement of multiple proteins is possible; however, it requires stripping membranes of antibody and then reprobing with a second antibody. Miniaturized alternatives to Western blot based on microfluidic or capillary electrophoresis have been developed that enable higher-throughput, automation, and greater mass sensitivity. In one approach, proteins are separated by electrophoresis on a microchip that is dragged along a polyvinylidene fluoride membrane so that as proteins exit the chip they are captured on the membrane for immunoassay. In this work, we improve this method to allow multiplexed protein detection. Multiple injections made from the same sample can be deposited in separate tracks so that each is probed with a different antibody. To further enhance multiplexing capability, the electrophoresis channel dimensions were optimized for resolution while keeping separation and blotting times to less than 8 min. Using a 15 μm deep × 50 μm wide × 8.6 cm long channel, it is possible to achieve baseline resolution of proteins that differ by 5% in molecular weight, e.g., ERK1 (44 kDa) from ERK2 (42 kDa). This resolution allows similar proteins detected by cross-reactive antibodies in a single track. We demonstrate detection of 11 proteins from 9 injections from a single Jurkat cell lysate sample consisting of 400 ng of total protein using this procedure. Thus, multiplexed Western blots are possible without cumbersome stripping and reprobing steps. PMID:27270033

  3. Acceleration of sea-ice Melt Addition Into the Northern North Atlantic

    NASA Astrophysics Data System (ADS)

    Cox, K. A.; Stanford, J. D.; McVicar, A. J.; Rohling, E. J.; Bacon, S.; Bolshaw, M.; de La Rosa, S.; Wilkinson, D.

    2008-12-01

    Observations over the last decade suggest both a thinning of the Arctic sea-ice cover (Rothrock et al., 1999; Laxon et al., 2003) and a dramatic reduction in its spatial extent (Comiso, 2002; Stroeve et al., 2005). We detect a positive change of ~10 ‰ between 1998 and 2005 in the stable oxygen isotope composition of the net freshwater component in the East Greenland Current (EGC) and East Greenland Coastal Current (EGCC), key carriers of freshened surface waters out of the Arctic. This isotopic signal is unique in the context of oxygen isotope measurements in the northern North Atlantic region dating back to the early 1960s. We show that this signal reflects a remarkable increase in the sea-ice melt water transport within these currents, to the equivalent of a net minimum volumetric loss of multi-year ('permanent') Arctic sea-ice of 10 ± 3% per decade. This independent measurement therefore complements and corroborates the sea-ice reduction inferred from satellite data, and places it in a longer-term multi-decadal context. Our findings suggest that a large proportion of the sea-ice meltwater resulting from the rapid reduction of Arctic sea-ice between 1998 and 2005 is exported from the Arctic via the EGC/EGCC into the northern North Atlantic. Additionally it appears that this sea-ice meltwater export is not in phase with atmospheric circulation regimes such as the North Atlantic Oscillation and the Arctic Oscillation. These findings could have important ramifications for the global thermohaline circulation (e.g. Rahmstorf, 2005).

  4. Additional Arctic observations improve weather and sea-ice forecasts for the Northern Sea Route

    PubMed Central

    Inoue, Jun; Yamazaki, Akira; Ono, Jun; Dethloff, Klaus; Maturilli, Marion; Neuber, Roland; Edwards, Patti; Yamaguchi, Hajime

    2015-01-01

    During ice-free periods, the Northern Sea Route (NSR) could be an attractive shipping route. The decline in Arctic sea-ice extent, however, could be associated with an increase in the frequency of the causes of severe weather phenomena, and high wind-driven waves and the advection of sea ice could make ship navigation along the NSR difficult. Accurate forecasts of weather and sea ice are desirable for safe navigation, but large uncertainties exist in current forecasts, partly owing to the sparse observational network over the Arctic Ocean. Here, we show that the incorporation of additional Arctic observations improves the initial analysis and enhances the skill of weather and sea-ice forecasts, the application of which has socioeconomic benefits. Comparison of 63-member ensemble atmospheric forecasts, using different initial data sets, revealed that additional Arctic radiosonde observations were useful for predicting a persistent strong wind event. The sea-ice forecast, initialised by the wind fields that included the effects of the observations, skilfully predicted rapid wind-driven sea-ice advection along the NSR. PMID:26585690

  5. Additional Arctic observations improve weather and sea-ice forecasts for the Northern Sea Route

    NASA Astrophysics Data System (ADS)

    Inoue, Jun; Yamazaki, Akira; Ono, Jun; Dethloff, Klaus; Maturilli, Marion; Neuber, Roland; Edwards, Patti; Yamaguchi, Hajime

    2015-11-01

    During ice-free periods, the Northern Sea Route (NSR) could be an attractive shipping route. The decline in Arctic sea-ice extent, however, could be associated with an increase in the frequency of the causes of severe weather phenomena, and high wind-driven waves and the advection of sea ice could make ship navigation along the NSR difficult. Accurate forecasts of weather and sea ice are desirable for safe navigation, but large uncertainties exist in current forecasts, partly owing to the sparse observational network over the Arctic Ocean. Here, we show that the incorporation of additional Arctic observations improves the initial analysis and enhances the skill of weather and sea-ice forecasts, the application of which has socioeconomic benefits. Comparison of 63-member ensemble atmospheric forecasts, using different initial data sets, revealed that additional Arctic radiosonde observations were useful for predicting a persistent strong wind event. The sea-ice forecast, initialised by the wind fields that included the effects of the observations, skilfully predicted rapid wind-driven sea-ice advection along the NSR.

  6. Additional Arctic observations improve weather and sea-ice forecasts for the Northern Sea Route.

    PubMed

    Inoue, Jun; Yamazaki, Akira; Ono, Jun; Dethloff, Klaus; Maturilli, Marion; Neuber, Roland; Edwards, Patti; Yamaguchi, Hajime

    2015-01-01

    During ice-free periods, the Northern Sea Route (NSR) could be an attractive shipping route. The decline in Arctic sea-ice extent, however, could be associated with an increase in the frequency of the causes of severe weather phenomena, and high wind-driven waves and the advection of sea ice could make ship navigation along the NSR difficult. Accurate forecasts of weather and sea ice are desirable for safe navigation, but large uncertainties exist in current forecasts, partly owing to the sparse observational network over the Arctic Ocean. Here, we show that the incorporation of additional Arctic observations improves the initial analysis and enhances the skill of weather and sea-ice forecasts, the application of which has socioeconomic benefits. Comparison of 63-member ensemble atmospheric forecasts, using different initial data sets, revealed that additional Arctic radiosonde observations were useful for predicting a persistent strong wind event. The sea-ice forecast, initialised by the wind fields that included the effects of the observations, skilfully predicted rapid wind-driven sea-ice advection along the NSR. PMID:26585690

  7. The responses of soil respiration to nitrogen addition in a temperate grassland in northern China.

    PubMed

    Luo, Qinpu; Gong, Jirui; Zhai, Zhanwei; Pan, Yan; Liu, Min; Xu, Sha; Wang, Yihui; Yang, Lili; Baoyin, Taoge-Tao

    2016-11-01

    Anthropogenic activities have increased nitrogen (N) inputs to grassland ecosystems. Knowledge of the impact of soil N availability on soil respiration (RS) is critical to understand soil carbon balances and their responses to global climate change. A 2-year field experiment was conducted to evaluate the response of RS to soil mineral N in a temperate grassland in northern China. RS, abiotic and biotic factors, and N mineralization were measured in the grassland, at rates of N addition ranging from 0 to 25gNm(-2)yr(-1). Annual and dormant-season RS ranged from 241.34 to 283.64g C m(-2) and from 61.34 to 83.84g C m(-2) respectively. High N application significantly increased RS, possibly due to increased root biomass and increased microbial biomass. High N treatment significantly increased soil NO3-N and inorganic N content compared with the control. The ratio of NO3-N to NH4-N and the N mineralization rate were significantly positively correlated with RS, but NH4-N was not correlated or negatively correlated with RS during the growing season. The temperature sensitivity of RS (Q10) was not significantly affected by N levels, and ranged from 1.90 to 2.20, but decreased marginally significantly at high N. RS outside the growing season is an important component of annual RS, accounting for 25.0 to 29.6% of the total. High N application indirectly stimulated RS by increasing soil NO3-N and net nitrification, thereby eliminating soil N limitations, promoting ecosystem productivity, and increasing soil CO2 efflux. Our results show the importance of distinguishing between NO3-N and NH4-N, as their impact on soil CO2 efflux differed. PMID:27396319

  8. Effects of Nitrogen Addition on Nitrogen Resorption in Temperate Shrublands in Northern China.

    PubMed

    Zhang, Jianhua; Li, He; Shen, Haihua; Chen, Yahan; Fang, Jingyun; Tang, Zhiyao

    2015-01-01

    Nutrient resorption from senescing leaves is a key mechanism of nutrient conservation for plants. The nutrient resorption efficiency is highly dependent on leaf nutrient status, species identity and soil nutrient availability. Nitrogen is a limiting nutrient in most ecosystems, it is widely reported that nitrogen resorption efficiency (NRE) was highly dependent on the soil nitrogen availability and vary with N deposition. The effects of nitrogen deposition on NRE and nitrogen concentration in green and senescing leaves have been well established for forests and grasslands; in contrast, little is known on how plants in shrublands respond to nitrogen deposition across the world. In this study, we conducted a two-year nitrogen addition manipulation experiment to explore the responses of nitrogen concentration in green and senescing leaves, and NRE of seven dominant species, namely, Vitex negundo, Wikstroemia chamaedaphne, Carex rigescens and Cleistogenes chinensis from the Vitex negundo community, and Spirea trilobata, Armeniaca sibirica, V. negundo, C. rigescens and Spodiopogon sibiricus from the Spirea trilobata community, to nitrogen deposition in two typical shrub communities of Mt. Dongling in northern China. Results showed that NRE varied remarkably among different life forms, which was lowest in shrubs, highest in grasses, and intermediate in forbs, implying that shrubs may be most capable of obtaining nitrogen from soil, grasses may conserve more nitrogen by absorption from senescing leaves, whereas forbs may adopt both mechanisms to compete for limited nitrogen supply from the habitats. As the N addition rate increases, N concentration in senescing leaves ([N]s) increased consistent from all species from both communities, that in green leaves ([N]g) increased for all species from the Vitex negundo community, while no significant responses were found for all species from the Spirea trilobata community; NRE decreased for all species except A. sibirica from the

  9. Effects of Nitrogen Addition on Nitrogen Resorption in Temperate Shrublands in Northern China

    PubMed Central

    Zhang, Jianhua; Li, He; Shen, Haihua; Chen, Yahan; Fang, Jingyun; Tang, Zhiyao

    2015-01-01

    Nutrient resorption from senescing leaves is a key mechanism of nutrient conservation for plants. The nutrient resorption efficiency is highly dependent on leaf nutrient status, species identity and soil nutrient availability. Nitrogen is a limiting nutrient in most ecosystems, it is widely reported that nitrogen resorption efficiency (NRE) was highly dependent on the soil nitrogen availability and vary with N deposition. The effects of nitrogen deposition on NRE and nitrogen concentration in green and senescing leaves have been well established for forests and grasslands; in contrast, little is known on how plants in shrublands respond to nitrogen deposition across the world. In this study, we conducted a two-year nitrogen addition manipulation experiment to explore the responses of nitrogen concentration in green and senescing leaves, and NRE of seven dominant species, namely, Vitex negundo, Wikstroemia chamaedaphne, Carex rigescens and Cleistogenes chinensis from the Vitex negundo community, and Spirea trilobata, Armeniaca sibirica, V. negundo, C. rigescens and Spodiopogon sibiricus from the Spirea trilobata community, to nitrogen deposition in two typical shrub communities of Mt. Dongling in northern China. Results showed that NRE varied remarkably among different life forms, which was lowest in shrubs, highest in grasses, and intermediate in forbs, implying that shrubs may be most capable of obtaining nitrogen from soil, grasses may conserve more nitrogen by absorption from senescing leaves, whereas forbs may adopt both mechanisms to compete for limited nitrogen supply from the habitats. As the N addition rate increases, N concentration in senescing leaves ([N]s) increased consistent from all species from both communities, that in green leaves ([N]g) increased for all species from the Vitex negundo community, while no significant responses were found for all species from the Spirea trilobata community; NRE decreased for all species except A. sibirica from the

  10. An automated rotisserie system for processing Western blots.

    PubMed

    Ma, P W K

    2002-01-01

    An apparatus to automate completely the processing of Western blots is described. The prototype is based on a popular rotisserie system design. The incubation chamber consists of an inner cylinder that rotates inside an outer cylinder (incubation chamber). The blot is contained in the inner cylinder. Two magnets are mounted at one end of the inner cylinder, and rotation of the inner cylinder is effected by two magnets mounted on a motor drive outside the incubation chamber. Movement of chemicals into and out of the incubation chamber is driven pneumatically, and the entire process is controlled by a computer. Processing a blot for chemiluminescent detection takes 7 h to complete without human intervention. The quality of the resulting image is comparable to or better than a blot using manual processing. In addition, the prototype is capable of re-collecting all three antisera for future use. PMID:12564602

  11. Problem-Solving Test: Southwestern Blotting

    ERIC Educational Resources Information Center

    Szeberényi, József

    2014-01-01

    Terms to be familiar with before you start to solve the test: Southern blotting, Western blotting, restriction endonucleases, agarose gel electrophoresis, nitrocellulose filter, molecular hybridization, polyacrylamide gel electrophoresis, proto-oncogene, c-abl, Src-homology domains, tyrosine protein kinase, nuclear localization signal, cDNA,…

  12. Additives

    NASA Technical Reports Server (NTRS)

    Smalheer, C. V.

    1973-01-01

    The chemistry of lubricant additives is discussed to show what the additives are chemically and what functions they perform in the lubrication of various kinds of equipment. Current theories regarding the mode of action of lubricant additives are presented. The additive groups discussed include the following: (1) detergents and dispersants, (2) corrosion inhibitors, (3) antioxidants, (4) viscosity index improvers, (5) pour point depressants, and (6) antifouling agents.

  13. Effects of Water and Nitrogen Addition on Species Turnover in Temperate Grasslands in Northern China

    PubMed Central

    Xu, Zhuwen; Wan, Shiqiang; Ren, Haiyan; Han, Xingguo; Li, Mai-He; Cheng, Weixin; Jiang, Yong

    2012-01-01

    Global nitrogen (N) deposition and climate change have been identified as two of the most important causes of current plant diversity loss. However, temporal patterns of species turnover underlying diversity changes in response to changing precipitation regimes and atmospheric N deposition have received inadequate attention. We carried out a manipulation experiment in a steppe and an old-field in North China from 2005 to 2009, to test the hypothesis that water addition enhances plant species richness through increase in the rate of species gain and decrease in the rate of species loss, while N addition has opposite effects on species changes. Our results showed that water addition increased the rate of species gain in both the steppe and the old field but decreased the rates of species loss and turnover in the old field. In contrast, N addition increased the rates of species loss and turnover in the steppe but decreased the rate of species gain in the old field. The rate of species change was greater in the old field than in the steppe. Water interacted with N to affect species richness and species turnover, indicating that the impacts of N on semi-arid grasslands were largely mediated by water availability. The temporal stability of communities was negatively correlated with rates of species loss and turnover, suggesting that water addition might enhance, but N addition would reduce the compositional stability of grasslands. Experimental results support our initial hypothesis and demonstrate that water and N availabilities differed in the effects on rate of species change in the temperate grasslands, and these effects also depend on grassland types and/or land-use history. Species gain and loss together contribute to the dynamic change of species richness in semi-arid grasslands under future climate change. PMID:22768119

  14. Streamlined Strategies to Better Visualize Southern Blotting

    ERIC Educational Resources Information Center

    Dean, Derek M.

    2012-01-01

    In this article, I describe an animated slideshow of Southern blotting that I have made freely available to other instructors. My hope is to provide a clear visualization of the logistics behind the technique so that instructors have a solid basis--as well as time freed up--to discuss its applications with students.

  15. Single cell-resolution western blotting.

    PubMed

    Kang, Chi-Chih; Yamauchi, Kevin A; Vlassakis, Julea; Sinkala, Elly; Duncombe, Todd A; Herr, Amy E

    2016-08-01

    This protocol describes how to perform western blotting on individual cells to measure cell-to-cell variation in protein expression levels and protein state. Like conventional western blotting, single-cell western blotting (scWB) is particularly useful for protein targets that lack selective antibodies (e.g., isoforms) and in cases in which background signal from intact cells is confounding. scWB is performed on a microdevice that comprises an array of microwells molded in a thin layer of a polyacrylamide gel (PAG). The gel layer functions as both a molecular sieving matrix during PAGE and a blotting scaffold during immunoprobing. scWB involves five main stages: (i) gravity settling of cells into microwells; (ii) chemical lysis of cells in each microwell; (iii) PAGE of each single-cell lysate; (iv) exposure of the gel to UV light to blot (immobilize) proteins to the gel matrix; and (v) in-gel immunoprobing of immobilized proteins. Multiplexing can be achieved by probing with antibody cocktails and using antibody stripping/reprobing techniques, enabling detection of 10+ proteins in each cell. We also describe microdevice fabrication for both uniform and pore-gradient microgels. To extend in-gel immunoprobing to gels of small pore size, we describe an optional gel de-cross-linking protocol for more effective introduction of antibodies into the gel layer. Once the microdevice has been fabricated, the assay can be completed in 4-6 h by microfluidic novices and it generates high-selectivity, multiplexed data from single cells. The technique is relevant when direct measurement of proteins in single cells is needed, with applications spanning the fundamental biosciences to applied biomedicine. PMID:27466711

  16. Non-additive effects of litter diversity on greenhouse gas emissions from alpine steppe soil in Northern Tibet

    PubMed Central

    Chen, Youchao; Sun, Jian; Xie, Fangting; Yan, Yan; Wang, Xiaodan; Cheng, Genwei; Lu, Xuyang

    2015-01-01

    While litter decomposition is a fundamental ecological process, previous studies have mainly focused on the decay of single species. In this study, we conducted a litter-mixing experiment to investigate litter diversity effects on greenhouse gas (GHG) emissions from an alpine steppe soil in Northern Tibet. Significant non-additive effects of litter diversity on GHG dynamics can be detected; these non-additive effects were the result of species composition rather than species richness. Synergistic effects were frequent for CO2 and N2O emissions, as they were found to occur in 70.5% and 47.1% of total cases, respectively; antagonistic effects on CH4 uptake predominated in 60.3% of the cases examined. The degree of synergism and antagonism may be significantly impacted by litter chemical traits, such as lignin and N, lignin:N ratio, and total phenols during decomposition (P < 0.05). In addition, the relationship between chemical traits and litter-mixing effects changed over incubation time. Our study provides an opportunity to gain insight into the relationship between litter diversity and soil ecological processes. The results indicate that higher plant diversity may generally enhance CO2 and N2O emissions while inhibiting CH4 uptake; meanwhile, the direction and strength of non-additive effects appear to be related to litter chemical traits. PMID:26634911

  17. Non-additive effects of litter diversity on greenhouse gas emissions from alpine steppe soil in Northern Tibet.

    PubMed

    Chen, Youchao; Sun, Jian; Xie, Fangting; Yan, Yan; Wang, Xiaodan; Cheng, Genwei; Lu, Xuyang

    2015-01-01

    While litter decomposition is a fundamental ecological process, previous studies have mainly focused on the decay of single species. In this study, we conducted a litter-mixing experiment to investigate litter diversity effects on greenhouse gas (GHG) emissions from an alpine steppe soil in Northern Tibet. Significant non-additive effects of litter diversity on GHG dynamics can be detected; these non-additive effects were the result of species composition rather than species richness. Synergistic effects were frequent for CO2 and N2O emissions, as they were found to occur in 70.5% and 47.1% of total cases, respectively; antagonistic effects on CH4 uptake predominated in 60.3% of the cases examined. The degree of synergism and antagonism may be significantly impacted by litter chemical traits, such as lignin and N, lignin:N ratio, and total phenols during decomposition (P < 0.05). In addition, the relationship between chemical traits and litter-mixing effects changed over incubation time. Our study provides an opportunity to gain insight into the relationship between litter diversity and soil ecological processes. The results indicate that higher plant diversity may generally enhance CO2 and N2O emissions while inhibiting CH4 uptake; meanwhile, the direction and strength of non-additive effects appear to be related to litter chemical traits. PMID:26634911

  18. BLOT II Ver.1.39

    SciTech Connect

    2003-06-03

    BLOT II is a graphic program for post-processing finite element analyses output in the EXODUS II database format. It is command driven with free-format input and can drive graphics devices supported by the Sandia Virtual Device Interface. BLOT produces mesh plots of the analysis output variables including deformed mesh plots, line contours, filled (painted) contours, vector plots of two/three variables (velocity vectors), and symbol plots of scalar variables (discrete cracks). Features include pathlines of analysis variables drawn on the mesh, element selection by material, element birth and death, multiple views combining several displays on each plot, symmetry mirroring, and node and element numbering. X-Y plots of the analysis variables include time vs. variable plots or variable vs. variable plots, and distance vs. variable plots at selected time stips where distance is the accumulated distance between pairs of nodes or element centers.

  19. BLOT II Ver.1.39

    2003-06-03

    BLOT II is a graphic program for post-processing finite element analyses output in the EXODUS II database format. It is command driven with free-format input and can drive graphics devices supported by the Sandia Virtual Device Interface. BLOT produces mesh plots of the analysis output variables including deformed mesh plots, line contours, filled (painted) contours, vector plots of two/three variables (velocity vectors), and symbol plots of scalar variables (discrete cracks). Features include pathlines of analysismore » variables drawn on the mesh, element selection by material, element birth and death, multiple views combining several displays on each plot, symmetry mirroring, and node and element numbering. X-Y plots of the analysis variables include time vs. variable plots or variable vs. variable plots, and distance vs. variable plots at selected time stips where distance is the accumulated distance between pairs of nodes or element centers.« less

  20. Two-dimensional southwestern blotting and characterization of transcription factors on-blot.

    PubMed

    Jiang, Daifeng; Jia, Yinshan; Zhou, YanWen; Jarrett, Harry W

    2009-07-01

    Two-dimensional Southwestern blotting (2D-SW) described here combines several steps. Proteins are separated by two-dimensional gel electrophoresis and transferred to nitrocellulose (NC) or polyvinylidene fluoride (PVDF) membrane. The blotted proteins are then partially renatured and probed with a specific radiolabeled oligonucleotide for Southwestern blotting (SW) analysis. The detected proteins are then processed by on-blot digestion and identified by LC-MS/MS analysis. A transcription factor, bound by a specific radiolabeled element, is thus characterized without aligning with protein spots on a gel. In this study, we systematically optimize conditions for 2D-SW and on-blot digestion. By quantifying the SW signal using a scintillation counter, the optimal conditions for SW were determined to be PVDF membrane, 0.5% PVP40 for membrane blocking, serial dilution of guanidine HCl for denaturing and renaturing proteins on the blot, and an SDS stripping buffer to remove radiation from the blot. By the quantification of the peptide yields using nano-ESI-MS analysis, the optimized conditions for on-blot digestions were found to be 0.5% Zwittergent 3-16 and 30% acetonitrile in trypsin digestion buffer. With the use of the optimized 2D-SW technique and on-blot digestion combined with HPLC-nano-ESI-MS/MS, a GFP-C/EBP model protein was successfully characterized from a bacterial extract, and native C/EBP beta was identified from 100 microg of HEK293 nuclear extract without any previous purification. PMID:19388704

  1. Western Blotting Inaccuracies with Unverified Antibodies: Need for a Western Blotting Minimal Reporting Standard (WBMRS)

    PubMed Central

    Gilda, Jennifer E.; Ghosh, Rajeshwary; Cheah, Jenice X.; West, Toni M.; Bodine, Sue C.; Gomes, Aldrin V.

    2015-01-01

    Western blotting is a commonly used technique in biological research. A major problem with Western blotting is not the method itself, but the use of poor quality antibodies as well as the use of different experimental conditions that affect the linearity and sensitivity of the Western blot. Investigation of some conditions that are commonly used and often modified in Western blotting, as well as some commercial antibodies, showed that published articles often fail to report critical parameters needed to reproduce the results. These parameters include the amount of protein loaded, the blocking solution and conditions used, the amount of primary and secondary antibodies used, the antibody incubation solutions, the detection method and the quantification method utilized. In the present study, comparison of ubiquitinated proteins in rat heart and liver samples showed different results depending on the antibody utilized. Validation of five commercial ubiquitin antibodies using purified ubiquitinated proteins, ubiquitin chains and free ubiquitin showed that these antibodies differ in their ability to detect free ubiquitin or ubiquitinated proteins. Investigating proteins modified with interferon-stimulated gene 15 (ISG15) in young and old rat hearts using six commercially available antibodies showed that most antibodies gave different semi-quantitative results, suggesting large variability among antibodies. Evidence showing the importance of the Western blot buffer and the concentration of antibody used is presented. Hence there is a critical need for comprehensive reporting of experimental conditions to improve the accuracy and reproducibility of Western blot analysis. A Western blotting minimal reporting standard (WBMRS) is suggested to improve the reproducibility of Western blot analysis. PMID:26287535

  2. Aboveground and belowground responses to nutrient additions and herbivore exclusion in Arctic tundra ecosystems in northern Alaska

    NASA Astrophysics Data System (ADS)

    Moore, J. C.; Gough, L.; Simpson, R.; Johnson, D. R.

    2011-12-01

    The Arctic has experienced significant increased regional warming over the past 30 years. Warming generally increases tundra soil nutrient availability by creating a more favorable environment for plant growth, decomposition and nutrient mineralization. Aboveground there has been a "greening" of the Arctic with increased net primary productivity (NPP), and an increase in woody vegetation. Concurrent with the changes aboveground has been an increase in root growth at lower depths and a loss of soil organic C (40 -100 g C m-2 yr-1). Given that arctic soils contain 14% of the global soil C pool, understanding the mechanisms behind shifts of this magnitude that are changing arctic soils from a net sink to a net source of atmospheric C is critical. We took an integrated multi-trophic level approach to examine how altering soil nutrients and mammalian herbivore activity affects vegetation, soil fauna, and microbial communities as well as soil physical characteristics in moist acidic (MAT) and dry heath (DH) tundra. Our work was conducted at the Arctic LTER site in northern Alaska. We sampled the nutrient (controls and annual N+P additions) and herbivore (controls and exclosures) manipulations established in 1996 after 10 years of treatment. Models that incorporated the biomass estimates from the field were used to characterize the trophic structure of the belowground food web and to estimate carbon flux among soil organisms and C-mineralization rates. Both MAT and DH exhibited significant increases in NPP and root growth and changes in vegetation structure with transitions from a mixed community to deciduous shrubs in MAT and from lichens to grasses and shrubs in DH, with nutrient additions and herbivore exclosures. Belowground responses to the treatments were dependent on ecosystem type, but exposed alterations in trophic structure that included changes in microbial biomass, the establishment of microbivorous enchytreaids, increases in root-feeding nematodes, and

  3. Antibody performance in western blot applications is context-dependent.

    PubMed

    Algenäs, Cajsa; Agaton, Charlotta; Fagerberg, Linn; Asplund, Anna; Björling, Lisa; Björling, Erik; Kampf, Caroline; Lundberg, Emma; Nilsson, Peter; Persson, Anja; Wester, Kenneth; Pontén, Fredrik; Wernérus, Henrik; Uhlén, Mathias; Ottosson Takanen, Jenny; Hober, Sophia

    2014-03-01

    An important concern for the use of antibodies in various applications, such as western blot (WB) or immunohistochemistry (IHC), is specificity. This calls for systematic validations using well-designed conditions. Here, we have analyzed 13 000 antibodies using western blot with lysates from human cell lines, tissues, and plasma. Standardized stratification showed that 45% of the antibodies yielded supportive staining, and the rest either no staining (12%) or protein bands of wrong size (43%). A comparative study of WB and IHC showed that the performance of antibodies is application-specific, although a correlation between no WB staining and weak IHC staining could be seen. To investigate the influence of protein abundance on the apparent specificity of the antibody, new WB analyses were performed for 1369 genes that gave unsupportive WBs in the initial screening using cell lysates with overexpressed full-length proteins. Then, more than 82% of the antibodies yielded a specific band corresponding to the full-length protein. Hence, the vast majority of the antibodies (90%) used in this study specifically recognize the target protein when present at sufficiently high levels. This demonstrates the context- and application-dependence of antibody validation and emphasizes that caution is needed when annotating binding reagents as specific or cross-reactive. WB is one of the most commonly used methods for validation of antibodies. Our data implicate that solely using one platform for antibody validation might give misleading information and therefore at least one additional method should be used to verify the achieved data. PMID:24403002

  4. BLOTS AND ALL: A HISTORY OF THE RORSCHACH INK BLOT TEST IN BRITAIN.

    PubMed

    Hubbard, Katherine; Hegarty, Peter

    2016-01-01

    Despite the easily recognizable nature of the Rorschach ink blot test very little is known about the history of the test in Britain. We attend to the oft-ignored history of the Rorschach test in Britain and compare it to its history in the US. Prior to the Second World War, Rorschach testing in Britain had attracted advocates and critiques. Afterward, the British Rorschach Forum, a network with a high proportion of women, developed around the Tavistock Institute in London and The Rorschach Newsletter. In 1968, the International Rorschach Congress was held in London but soon after the group became less exclusive, and fell into decline. A comparative account of the Rorschach in Britain demonstrates how different national institutions invested in the 'projective hypothesis' according to the influence of psychoanalysis, the adoption of a nationalized health system, and the social positioning of 'others' throughout the twentieth century. In comparing and contrasting the history of the Rorschach in Britain and the US, we decentralize and particularize the history of North American Psychology. PMID:26924673

  5. Soil microbial responses to forest floor litter manipulation and nitrogen addition in a mixed-wood forest of northern China.

    PubMed

    Sun, Xiao-Lu; Zhao, Jing; You, Ye-Ming; Jianxin Sun, Osbert

    2016-01-01

    Changes in litterfall dynamics and soil properties due to anthropogenic or natural perturbations have important implications to soil carbon (C) and nutrient cycling via microbial pathway. Here we determine soil microbial responses to contrasting types of litter inputs (leaf vs. fine woody litter) and nitrogen (N) deposition by conducting a multi-year litter manipulation and N addition experiment in a mixed-wood forest. We found significantly higher soil organic C, total N, microbial biomass C (MBC) and N (MBN), microbial activity (MR), and activities of four soil extracellular enzymes, including β-glucosidase (BG), N-acetyl-β-glucosaminidase (NAG), phenol oxidase (PO), and peroxidase (PER), as well as greater total bacteria biomass and relative abundance of gram-negative bacteria (G-) community, in top soils of plots with presence of leaf litter than of those without litter or with presence of only fine woody litter. No apparent additive or interactive effects of N addition were observed in this study. The occurrence of more labile leaf litter stimulated G-, which may facilitate microbial community growth and soil C stabilization as inferred by findings in literature. A continued treatment with contrasting types of litter inputs is likely to result in divergence in soil microbial community structure and function. PMID:26762490

  6. Soil microbial responses to forest floor litter manipulation and nitrogen addition in a mixed-wood forest of northern China

    PubMed Central

    Sun, Xiao-Lu; Zhao, Jing; You, Ye-Ming; Jianxin Sun, Osbert

    2016-01-01

    Changes in litterfall dynamics and soil properties due to anthropogenic or natural perturbations have important implications to soil carbon (C) and nutrient cycling via microbial pathway. Here we determine soil microbial responses to contrasting types of litter inputs (leaf vs. fine woody litter) and nitrogen (N) deposition by conducting a multi-year litter manipulation and N addition experiment in a mixed-wood forest. We found significantly higher soil organic C, total N, microbial biomass C (MBC) and N (MBN), microbial activity (MR), and activities of four soil extracellular enzymes, including β-glucosidase (BG), N-acetyl-β-glucosaminidase (NAG), phenol oxidase (PO), and peroxidase (PER), as well as greater total bacteria biomass and relative abundance of gram-negative bacteria (G-) community, in top soils of plots with presence of leaf litter than of those without litter or with presence of only fine woody litter. No apparent additive or interactive effects of N addition were observed in this study. The occurrence of more labile leaf litter stimulated G-, which may facilitate microbial community growth and soil C stabilization as inferred by findings in literature. A continued treatment with contrasting types of litter inputs is likely to result in divergence in soil microbial community structure and function. PMID:26762490

  7. Quantitative autoradiography of dot blots using a microwell densitometer

    SciTech Connect

    Ross, P.M.; Woodley, K.; Baird, M. )

    1989-07-01

    We have established conditions for the quantitation of DNA hybridization by reading dot blot autoradiographs with a microwell plate densitometer. This method is more convenient, as accurate, and more sensitive than counting the spots in a liquid scintillation counter.

  8. IDENTIFICATION OF IMMUNOGENS OF 'MYCOPLASMA PNEUMONIAE' BY PROTEIN BLOTTING

    EPA Science Inventory

    Proteins of Mycoplasma pneumoniae were separated by SDS-polyacrylamide gel electrophoresis and transferred to a nitrocellulose sheet by blotting. Sera obtained from infected hamsters and immunized rabbits were then incubated with the nitrocellulose strips. Proteins which are capa...

  9. Optimized semi-quantitative blot analysis in infection assays using the Stain-Free technology.

    PubMed

    Zeitler, Anna F; Gerrer, Katrin H; Haas, Rainer; Jiménez-Soto, Luisa F

    2016-07-01

    Western blots are a commonly used method for protein detection and quantification in biological samples. Compensation of loading variations is achieved by housekeeping protein (HKP) normalization and/or total protein normalization (TPN). However, under infection conditions, HKP normalization, traditionally used in cell biology for quantification of western blots, can be problematic. Binding of microbes to target cells via specific receptors can induce signal transduction events resulting in drastic changes in the level of expression of HKPs. Additionally, samples collected after infection assays will include cellular and microbial proteins altering the analysis with TPN. Here we demonstrate under experimental infection conditions, how a reliable semi-quantitative analysis of proteins in western blots can be achieved using the Stain-Free technology. PMID:27150675

  10. Western Blot of Stained Proteins from Dried Polyacrylamide Gels

    NASA Technical Reports Server (NTRS)

    Gruber, Claudia; Stan-Lotter, Helga

    1996-01-01

    Western blotting of proteins is customarily performed following their separation on polyacrylamide gels, either prior to staining (1) or, as recently reported, following staining (2). We describe here Western blotting with stained gels, which had been dried and some of which had been stored for years. This procedure permits immunological analysis of proteins, to which antisera may have become available only later, or where the application of newly developed sensitive detection methods is desired. Once rehydration of the gels is achieved, proteins can be-transferred to blotting membranes by any appropriate protocol. Proteins stained with Coomassie Blue have to be detected with a non-chromogenic method, such as the film-based enhanced chemiluminescence (ECL)2) procedure (3). Silver stained proteins, which transfer in the colorless form, may be visualized by any detection method, although, because of the usually very low amounts of proteins, detection by ECL is preferable. Blotting of stained proteins from rehydrated gels is as rapid and as quantitative as from freshly prepared gels, in contrast to blotting from wet stained gels, which requires extensive washing and results in low transfer efficiency (2). Together with a photographic record of the gel pattern, unambiguous identification of immunoreactive proteins from complex mixtures is possible. Some further applications of this work are discussed.

  11. Genetic relatedness of orbiviruses by RNA-RNA blot hybridization

    SciTech Connect

    Bodkin, D.K.

    1985-01-01

    RNA-RNA blot hybridization was developed in order to identify type-specific genes among double-stranded (ds) RNA viruses, to assess the genetic relatedness of dsRNA viruses and to classify new strains. Viral dsRNA segments were electrophoresed through 10% polyacrylamide gels, transferred to membranes, and hybridized to (5'/sup 32/P)-pCp labeled genomic RNA from a related strain. Hybridization was performed at 52/sup 0/C, 50% formamide, 5X SSC. Under these conditions heterologous RNA species must share greater than or equal to 74% sequence homology in order to form stable dsRNA hybrids. Cognate genes of nine members of the Palyam serogroup of orbiviruses were identified and their sequence relatedness to the prototype. Palyam virus, was determined. Reciprocal blot hybridizations were performed using radiolabeled genomic RNA of all members of the Palyam serogroup. Unique and variant genes were identified by lack of cross-homology or by weak homology between segments. Since genes 2 and 6 exhibited the highest degree of sequence variability, response to the vertebrate immune system may be a major cause of sequence divergence among members of a single serogroup. Changuinola serogroup isolates were compared by dot-blot hybridization, while Colorado tick fever (CTF) serogroup isolates were compared by the RNA-RNA blot hybridization procedure described for reovirus and Palyam serogroup isolates. Preliminary blot hybridization data were also obtained on the relatedness of members of different Orbivirus serogroups.

  12. Improved semiquantitative Western blot technique with increased quantification range.

    PubMed

    Heidebrecht, F; Heidebrecht, A; Schulz, I; Behrens, S-E; Bader, A

    2009-06-30

    With the development of new interdisciplinary fields such as systems biology, the quantitative analysis of protein expression in biological samples gains more and more importance. Although the most common method for this is ELISA, Western blot also has advantages: The separation of proteins by size allows the evaluation of only specifically bound protein. This work examines the Western blot signal chain, determines some of the parameters relevant for quantitative analysis and proposes a mathematical model of the reaction kinetics. Using this model, a semiquantitative Western blot method for simultaneous quantification of different proteins using a hyperbolic calibration curve was developed. A program was written for the purpose of hyperbolic regression that allows quick determination of the calibration curve coefficients. This program can be used also for approximation of calibration curves in other applications such as ELISA, BCA or Bradford assays. PMID:19351538

  13. Detection of Blotted Proteins: Not All Blockers Are Created Equal.

    PubMed

    Kothari, Vishal; Mathews, Suresh T

    2015-01-01

    Western blotting is a standard analytical technique for detection of proteins. It is dependent on a number of components; from the specificity of the primary antibody to the reduction of competing biomolecules present in the assay. Blocking agents are a critical component for western blotting protocols as these diminish nonspecific binding by blocking off-target sites on the membrane. A variety of blocking agents are available and these are selected in an empirical manner, as no single blocker is compatible with every system. The best blocking agent and method for any particular assay will be an optimized but not absolute choice. Here, we describe characteristics of the most common blocking agents used in western blotting and discuss their advantages and disadvantages. PMID:26139251

  14. Effects of Reusing Gel Electrophoresis and Electrotransfer Buffers on Western Blotting

    PubMed Central

    Omotola, Oluwabukola B.; Heda, Rajiv P.; Avery, Jamie

    2016-01-01

    SDS-PAGE and Western blotting are 2 of the most commonly used biochemical methods for protein analysis. Proteins are electrophoretically separated based on their MWs by SDS-PAGE and then electrotransferred to a solid membrane surface for subsequent protein-specific analysis by immunoblotting, a procedure commonly known as Western blotting. Both of these procedures use a salt-based buffer, with the latter procedure consisting of methanol as an additive known for its toxicity. Previous reports present a contradictory view in favor or against reusing electrotransfer buffer, also known as Towbin’s transfer buffer (TTB), with an aim to reduce the toxic waste. In this report, we present a detailed analysis of not only reusing TTB but also gel electrophoresis buffer (EB) on proteins of low to high MW range. Our results suggest that EB can be reused for at least 5 times without compromising the electrophoretic separation of mixture of proteins in an MW standard, BSA, and crude cell lysates. Additionally, reuse of EB did not affect the quality of subsequent Western blots. Successive reuse of TTB, on the other hand, diminished the signal of proteins of different MWs in a protein standard and a high MW membrane protein cystic fibrosis transmembrane-conductance regulator (CFTR) in Western blotting. PMID:27582639

  15. Effects of Reusing Gel Electrophoresis and Electrotransfer Buffers on Western Blotting.

    PubMed

    Heda, Ghanshyam D; Omotola, Oluwabukola B; Heda, Rajiv P; Avery, Jamie

    2016-09-01

    SDS-PAGE and Western blotting are 2 of the most commonly used biochemical methods for protein analysis. Proteins are electrophoretically separated based on their MWs by SDS-PAGE and then electrotransferred to a solid membrane surface for subsequent protein-specific analysis by immunoblotting, a procedure commonly known as Western blotting. Both of these procedures use a salt-based buffer, with the latter procedure consisting of methanol as an additive known for its toxicity. Previous reports present a contradictory view in favor or against reusing electrotransfer buffer, also known as Towbin's transfer buffer (TTB), with an aim to reduce the toxic waste. In this report, we present a detailed analysis of not only reusing TTB but also gel electrophoresis buffer (EB) on proteins of low to high MW range. Our results suggest that EB can be reused for at least 5 times without compromising the electrophoretic separation of mixture of proteins in an MW standard, BSA, and crude cell lysates. Additionally, reuse of EB did not affect the quality of subsequent Western blots. Successive reuse of TTB, on the other hand, diminished the signal of proteins of different MWs in a protein standard and a high MW membrane protein cystic fibrosis transmembrane-conductance regulator (CFTR) in Western blotting. PMID:27582639

  16. [Contribution of Western blotting to the diagnosis of hydatidosis].

    PubMed

    Makni, F; Hachicha, L; Mseddi, F; Hammami, H; Cheikhrouhou, F; Sellami, H; Sellami, A; Mzali, R; Boujelbène, S; Rebaï, R; Beyrouti, I; Ayadi, A

    2007-08-01

    The aim of this study is to evaluate the contribution of the immunoWesternblot for the diagnosis and the post surgical follow-up of the hydatidosis. 71 sera from patients with hydatidosis confirmed by surgery were studied. All had a negative hydatic serology by screening tests (enzyme-linked immunosorbent assay, hemagglutination, electrosyneresis). 12 patients with sera in pre and post operative were monitored for 2 years. The Echinococcus Western blot IgG permitted to rectify the diagnosis of hydatidosis in 67.6 %. The rate of positivity was 100 % for the multivesicular liver cysts, 60 % for the young cysts and 50 % for the calcified cysts. Western blot permitted to rectify the diagnosis of lung cysts in 62.5 % of cases and in 50 % of cranial-spinal localizations. Analysis of Western Blot evolution in the 12 patients followed in pre and post-surgical revealed the disappearance of the bands 16, 18 and 26-28kDa in 8 month in the 8 patients with complete exeresis. This study proved the value added of Western blot compared to the other traditional techniques for the immunodiagnostic and the post-surgical monitoring of hydatidosis. PMID:17824307

  17. Better management of Western blotting results using professional photo management software.

    PubMed

    Iorio-Morin, Christian; Germain, Pascale; Parent, Jean-Luc

    2013-04-01

    Western blotting is a proven technique essential to a significant proportion of molecular biology projects. However, as results accumulate over the years, managing data can become daunting. Recognizing that the needs of a scientist working with Western blotting results are conceptually the same as those of a professional photographer managing a summer's worth of wedding photos, we report here a new workflow for managing Western blotting results using professional photo management software. The workflow involves (i) scanning all film-based results; (ii) importing the scans into the software; (iii) processing the scans; (iv) tagging the files with metadata, and (v) creating appropriate "smart-albums." Advantages of this system include space savings (both on our hard drives and on our desks), safer archival, quicker access, and easier sharing of the results. In addition, metadata-based workflows improve cross-experiment discovery and enable questions like "show me all blots labelled with antibody X" or "show me all experiments featuring protein Y". As project size and breadth increase, workflows delegating results management to the computer will become more and more important so that scientists can keep focussing on science. PMID:23404762

  18. Development of a heat-mediated protein blotting method.

    PubMed

    O'Sullivan, Jack; McMahon, Hilary E M

    2016-04-15

    Western blotting is a significant tool employed for the detection of cell proteins. High-molecular-weight proteins have proven a challenge to detect by western blotting, but proteins even of 100 KDa can still present difficulties in detection. This work reports the development of a heat transfer method that is suitable for both low- and high-molecular-weight proteins. The procedure involves the use of a constant temperature at 78 °C in a dedicated heat transfer module. Through the use of this protocol the neuronal adaptor protein X11α (120 KDa), which prior to this methodology was undetectable endogenously in the neuroblastoma cell line (N2a), was successfully detected in the N2a cell line. The procedure provides a reproducible protocol that can be adapted for other high-molecular-weight proteins, and it provides the advantage that low-molecular-weight proteins are not sacrificed by the methodology. PMID:26869081

  19. Lipid A binding proteins in macrophages detected by ligand blotting

    SciTech Connect

    Hampton, R.Y.; Golenbock, D.T.; Raetz, C.R.H.

    1987-05-01

    Endotoxin (LPS) stimulates a variety of eukaryotic cells. These actions are involved in the pathogenesis of Gram-negative septicemia. The site of action of the LPS toxic moiety, lipid A (LA), is unclear. Their laboratory has previously identified a bioactive LA precursor lipid IV/sub A/, which can be enzymatically labeled with /sup 32/P/sub i/ (10/sup 9/ dpm/nmole) and purified (99%). They now show that this ligand binds to specific proteins immobilized on nitrocellulose (NC) from LPS-sensitive RAW 264.7 cultured macrophages. NC blots were incubated with (/sup 32/P)-IV/sub A/ in a buffer containing BSA, NaCl, polyethylene glycol, and azide. Binding was assessed using autoradiography or scintillation counting. Dot blot binding of the radioligand was inhibited by excess cold IV/sub A/, LA, or ReLPS but not by phosphatidylcholine, cardiolipin, phosphatidylinositol, or phosphatidic acid. Binding was trypsin-sensitive and dependent on protein concentration. Particulate macrophage proteins were subjected to SDS-PAGE and then electroblotted onto NC. Several discrete binding proteins were observed. Identical treatment of fetal bovine serum or molecular weight standards revealed no detectable binding. By avoiding high nonspecific binding of intact membranes, this ligand blotting assay may be useful in elucidating the molecular actions of LPS.

  20. Relative impact of previous disturbance history on the likelihood of additional disturbance in the Northern United States Forest Service USFS Region

    NASA Astrophysics Data System (ADS)

    Hernandez, A. J.

    2015-12-01

    The Landsat archive is increasingly being used to detect trends in the occurrence of forest disturbance. Beyond information about the amount of area affected, forest managers need to know if and how disturbance regimes change. The National Forest System (NFS) has developed a comprehensive plan for carbon monitoring that requires a detailed temporal mapping of forest disturbances across 75 million hectares. A long-term annual time series that shows the timing, extent, and type of disturbance beginning in 1990 and ending in 2011 has been prepared for several USFS Regions, including the Northern Region. Our mapping starts with an automated detection of annual disturbances using a time series of historical Landsat imagery. Automated detections are meticulously inspected, corrected and labeled using various USFS ancillary datasets. The resulting maps of verified disturbance show the timing and types are fires, harvests, insect activity, disease, and abiotic (wind, drought, avalanche) damage. Also, the magnitude of each change event is modeled in terms of the proportion of canopy cover lost. The sequence of disturbances for every pixel since 1990 has been consistently mapped and is available across the entirety of NFS. Our datasets contain sufficient information to describe the frequency of stand replacement, as well as how often disturbance results in only a partial loss of canopy. This information provides empirical insight into how an initial disturbance may predispose a stand to further disturbance, and it also show a climatic signal in the occurrence of processes such as fire and insect epidemics. Thus, we have the information to model the likelihood of occurrence of certain disturbances after a given event (i.e. if we have a fire in the past what does that do to the likelihood of occurrence of insects in the future). Here, we explore if previous disturbance history is a reliable predictor of additional disturbance in the future and we present results of applying

  1. Differentiation of larva migrans caused by Baylisascaris procyonis and Toxocara species by Western blotting.

    PubMed

    Dangoudoubiyam, Sriveny; Kazacos, Kevin R

    2009-11-01

    Baylisascaris procyonis and Toxocara species are two important causes of larva migrans in humans. Larva migrans caused by Toxocara spp. is well known and is diagnosed serologically by enzyme immunoassay. Over a dozen cases of larva migrans and associated eosinophilic encephalitis caused by B. procyonis have also been reported, and at least a dozen additional cases are known. An enzyme-linked immunosorbent assay (ELISA) using the excretory-secretory (ES) antigen of B. procyonis larvae is currently being used in our laboratory as an aid in the diagnosis of this infection in humans. Clinically affected individuals show very high reactivity (measured as the optical density) on this ELISA; however, a one-way cross-reactivity with Toxocara spp. has been observed. As an approach to differentiate these two infections based on serology, we performed Western blots, wherein the B. procyonis ES antigen was reacted with serum samples from individuals known to be positive for either Toxocara spp. or B. procyonis larva migrans. Western blot results showed that B. procyonis antigens of between 30 and 45 kDa were specifically identified only by the sera from individuals with Baylisascaris larva migrans, thus allowing for differentiation between the two infections. This included human patient serum samples submitted for serologic testing, as well as sera from rabbits experimentally infected with B. procyonis. When used in conjunction with the ELISA, Western blotting could be an efficient tool for diagnosis of this infection in humans. PMID:19741091

  2. Selection of a Clostridium perfringens type D epsilon toxin producer via dot-blot test.

    PubMed

    Gonçalves, Luciana A; Lobato, Zélia I P; Silva, Rodrigo O S; Salvarani, Felipe M; Pires, Prhiscylla S; Assis, Ronnie A; Lobato, Francisco C F

    2009-11-01

    Clostridium perfringens type D produces enterotoxemia, an enteric disease in ruminants, also known as pulpy kidney disease. Caused by epsilon toxin, enterotoxemia is a major exotoxin produced by this microorganism. Epsilon toxin is also the main component of vaccines against this enteric disorder. In this study, a standardized dot-blot was used to choose strains of C. perfringens type D that are producers of epsilon toxin. Clones producing epsilon toxin were chosen by limiting dilution; after three passages, lethal minimum dose titers were determined by soroneutralization test in mice. These clones produced epsilon toxin 240 times more concentrated than the original strain. The presence of the epsilon toxin gene (etx) was verified by polymerase chain reaction. All clones were positive, including those determined to be negative by dot-blot tests, suggesting that mechanisms in addition to the presence of the etx gene can influence toxin production. The dot-blot test was efficient for the selection of toxigenic colonies of C. perfringens type D and demonstrated that homogeneous populations selected from toxigenic cultures produce higher titers of epsilon toxin. PMID:19779698

  3. The detection of DNA-binding proteins by protein blotting.

    PubMed Central

    Bowen, B; Steinberg, J; Laemmli, U K; Weintraub, H

    1980-01-01

    A method, called "protein blotting," for the detection of DNA-binding proteins is described. Proteins are separated on an SDA-polyacrylamide gel. The gel is sandwiched between 2 nitrocellulose filters and the proteins allowed to diffuse out of the gel and onto the filters. The proteins are tightly bound to each filter, producing a replica of the original gel pattern. The replica is used to detect DNA-binding proteins, RNA-binding proteins or histone-binding proteins by incubation of the filter with [32P]DNA, [125I]RNA, or [125I] histone. Evidence is also presented that specific protein-DNA interactions may be detected by this technique; under appropriate conditions, the lac repressor binds only to DNA containing the lac operator. Strategies for the detection of specific protein-DNA interactions are discussed. Images PMID:6243775

  4. PCR versus Southern blot detection of somatic mosaicism in fragile X syndrome

    SciTech Connect

    Mueller, O.T.; Amar, M.J.A.; Gallardo, L.A.; Kousseff, B.G.

    1994-09-01

    The incidence of somatic mosaicism in males with fragile X syndrome has been reported to be as high as 17% of all clinically affected males. Mosaic cases usually do not show the cytogenetic fragile site at Xq27.3 and generally are fully affected according to the clinical criteria, although some subjects show somewhat milder symptoms. Detection of mosaicism relies on the identification of multiple distinct sizes of the CGG size anomaly in the 5{prime} untranslated exon of the FMR-1 gene. Some alleles identified are of a premutation size with trinucleotides numbering 50 to 80 X CGG. In addition, there is a greatly expanded allele with well over 200 copies of the CGG repeat detected. We screened 314 subjects for fragile X syndrome over a three year period. Cases were routinely screened by Southern blotting using the StB12.3 probe as well as by polymerase chain reaction. The PCR amplification products were electrophoresed in agarose, blotted and hybridized with a (CGG){sub 5} oligonucleotide followed by chemiluminescent detection. Seventeen males and 16 females were identified with a CGG expansion, including two males with premutations with repeat sizes of between 50 and 100 CGG trinucleotides and three cases exhibiting somatic mosaicism. The mosaic cases had both a premutation-sized allele and one or more expanded alleles of over 250 CGG copies. The mosaic cases were usually undetected with Southern blotting but easily identified with this PCR protocol. The relative proportion of the expanded allele as determined by scanning densitometry were 70%, 35%, and 5% in the three cases. All three cases were cytogenetically negative. The clinical severity of the mosaic cases was variable, with symptoms ranging from severe MR with most of the physical stigmata to mild learning disability. In our experience, Southern blotting allows more accurate sizing of the expanded allele; however, PCR is essential to identify cases that exhibit mosaicism.

  5. Microfluidic Western Blotting of Low-Molecular-Mass Proteins

    PubMed Central

    2015-01-01

    We describe a microfluidic Western blot assay (μWestern) using a Tris tricine discontinuous buffer system suitable for analyses of a wide molecular mass range (6.5–116 kDa). The Tris tricine μWestern is completed in an enclosed, straight glass microfluidic channel housing a photopatterned polyacrylamide gel that incorporates a photoactive benzophenone methacrylamide monomer. Upon brief ultraviolet (UV) light exposure, the hydrogel toggles from molecular sieving for size-based separation to a covalent immobilization scaffold for in situ antibody probing. Electrophoresis controls all assay stages, affording purely electronic operation with no pumps or valves needed for fluid control. Electrophoretic introduction of antibody into and along the molecular sieving gel requires that the probe must traverse through (i) a discontinuous gel interface central to the transient isotachophoresis used to achieve high-performance separations and (ii) the full axial length of the separation gel. In-channel antibody probing of small molecular mass species is especially challenging, since the gel must effectively sieve small proteins while permitting effective probing with large-molecular-mass antibodies. To create a well-controlled gel interface, we introduce a fabrication method that relies on a hydrostatic pressure mismatch between the buffer and polymer precursor solution to eliminate the interfacial pore-size control issues that arise when a polymerizing polymer abuts a nonpolymerizing polymer solution. Combined with a new swept antibody probe plug delivery scheme, the Tris tricine μWestern blot enables 40% higher separation resolution as compared to a Tris glycine system, destacking of proteins down to 6.5 kDa, and a 100-fold better signal-to-noise ratio (SNR) for small pore gels, expanding the range of applicable biological targets. PMID:25268977

  6. Microfluidic Western blotting of low-molecular-mass proteins.

    PubMed

    Gerver, Rachel E; Herr, Amy E

    2014-11-01

    We describe a microfluidic Western blot assay (μWestern) using a Tris tricine discontinuous buffer system suitable for analyses of a wide molecular mass range (6.5-116 kDa). The Tris tricine μWestern is completed in an enclosed, straight glass microfluidic channel housing a photopatterned polyacrylamide gel that incorporates a photoactive benzophenone methacrylamide monomer. Upon brief ultraviolet (UV) light exposure, the hydrogel toggles from molecular sieving for size-based separation to a covalent immobilization scaffold for in situ antibody probing. Electrophoresis controls all assay stages, affording purely electronic operation with no pumps or valves needed for fluid control. Electrophoretic introduction of antibody into and along the molecular sieving gel requires that the probe must traverse through (i) a discontinuous gel interface central to the transient isotachophoresis used to achieve high-performance separations and (ii) the full axial length of the separation gel. In-channel antibody probing of small molecular mass species is especially challenging, since the gel must effectively sieve small proteins while permitting effective probing with large-molecular-mass antibodies. To create a well-controlled gel interface, we introduce a fabrication method that relies on a hydrostatic pressure mismatch between the buffer and polymer precursor solution to eliminate the interfacial pore-size control issues that arise when a polymerizing polymer abuts a nonpolymerizing polymer solution. Combined with a new swept antibody probe plug delivery scheme, the Tris tricine μWestern blot enables 40% higher separation resolution as compared to a Tris glycine system, destacking of proteins down to 6.5 kDa, and a 100-fold better signal-to-noise ratio (SNR) for small pore gels, expanding the range of applicable biological targets. PMID:25268977

  7. Additions to Philippine Slender Skinks of the Brachymeles bonitae Complex (Reptilia: Squamata: Scincidae) II: a new species from the northern Philippines.

    PubMed

    Siler, Cameron D; Davis, Drew R; Freitas, Elyse S; Huron, Nicholas A; Geheber, Aaron D; Watters, Jessa L; Penrod, Michelle L; Papeș, Monica; Amrein, Andrew; Anwar, Alyssa; Cooper, Dontae; Hein, Tucker; Manning, Annalisa; Patel, Neeral; Pinaroc, Lauren; Diesmos, Arvin C; Diesmos, Mae L; Oliveros, Carl H; Brown, Rafe M

    2016-01-01

    We describe a new digitless scincid lizard of the genus Brachymeles from northern Luzon and Camiguin Norte islands in the Philippines. This species belongs to the Brachymeles bonitae Complex, and both molecular and morphological data confirm that this species is distinct from all other congeners. Formerly considered to be a single widespread species, this group of species has been the focus of recent systematic reviews. Here we describe a new species in the B. bonitae Complex, recognized currently to constitute five species. Brachymeles ilocandia sp. nov. is the second digitless and the seventeenth non-pentadactyl species in genus. The description of this species brings the total number of species in the genus to 40, and provides new insight into unique distribution patterns of species of the northern Philippines. PMID:27395649

  8. Blotting Assisted by Heating and Solvent Extraction for DESI-MS Imaging

    NASA Astrophysics Data System (ADS)

    Cabral, Elaine C.; Mirabelli, Mario F.; Perez, Consuelo J.; Ifa, Demian R.

    2013-06-01

    Imprints of potato sprout ( Solanum tuberosum L.), gingko leaves (Gingko biloba L. ) and strawberries (Fragaria x ananassa Duch. ) were successfully imaged by desorption electrospray ionization mass spectrometry (DESI-MS) on TLC plates through blotting assisted by heating and/or solvent extraction. Ion images showing the distribution of significant compounds such as glycoalkaloid toxins in potato sprout, ginkgolic acids and flavonoids in ginkgo leaves, and sugars and anthocyanidin in strawberry were obtained. Practical implications of this work include analysis of a wide range of irregular or soft materials by different imprinting conditions without requiring the addition of matrices or use of specific kinds of surfaces.

  9. Blot overlays with 32P-labeled fusion proteins.

    PubMed

    Zhao, Z; Lim, L; Manser, E

    2001-07-01

    Proteins labeled with 32P can be used as sensitive "prime" in blot overlays to detect binding proteins or domains. Small G-protein Ras can bind GTP with extremely high affinity (Kd approximately 10(-11)-10(-12) M) in the presence of Mg2+. We have taken advantage of this property of Ras to develop a vector that expresses proteins of interest such as glutathione S-transferase (GST)/Ras fusion proteins for noncovalent labeling with [gamma-32P]GTP. The labeling efficiency of this method is >60% and involves a single short incubation step. We have previously identified several binding proteins for the second SH3 domain of the adaptor Nck using this method. Here we illustrate the overlay method using the GST/Ras system and compare results with the SH3 domain labeled by phosphorylation with [gamma-32P]ATP. Both methods are similarly specific and sensitive; however, we show that signals are dependent primarily on GST-mediated probe dimerization. These dimeric probes allow a more stable probe-target complex similar to immunoglobulin interactions, thus significantly improving the sensitivity of the technique. PMID:11403569

  10. Characterization of Nora Virus Structural Proteins via Western Blot Analysis

    PubMed Central

    Ericson, Brad L.; Carlson, Darby J.

    2016-01-01

    Nora virus is a single stranded RNA picorna-like virus with four open reading frames (ORFs). The coding potentials of the ORFs are not fully characterized, but ORF3 and ORF4 are believed to encode the capsid proteins (VP3, VP4a, VP4b, and VP4c) comprising the virion. To determine the polypeptide composition of Nora virus virions, polypeptides from purified virus were compared to polypeptides detected in Nora virus infected Drosophila melanogaster. Nora virus was purified from infected flies and used to challenge mice for the production of antisera. ORF3, ORF4a, ORF4b, and ORF4c were individually cloned and expressed in E. coli; resultant recombinant proteins purified and were used to make monospecific antisera. Antisera were evaluated via Western blot against whole virus particles and Nora virus infected fly lysates. Viral purification yielded two particle types with densities of ~1.31 g/mL (empty particles) and ~1.33 g/mL (complete virions). Comparison of purified virus polypeptide composition to Nora virus infected D. melanogaster lysate showed the number of proteins in infected cell lysates is less than purified virus. Our results suggest the virion is composed of 6 polypeptides, VP3, VP4a, two forms of VP4b, and two forms of VP4c. This polypeptide composition is similar to other small RNA insect viruses. PMID:27298753

  11. Characterization of Nora Virus Structural Proteins via Western Blot Analysis.

    PubMed

    Ericson, Brad L; Carlson, Darby J; Carlson, Kimberly A

    2016-01-01

    Nora virus is a single stranded RNA picorna-like virus with four open reading frames (ORFs). The coding potentials of the ORFs are not fully characterized, but ORF3 and ORF4 are believed to encode the capsid proteins (VP3, VP4a, VP4b, and VP4c) comprising the virion. To determine the polypeptide composition of Nora virus virions, polypeptides from purified virus were compared to polypeptides detected in Nora virus infected Drosophila melanogaster. Nora virus was purified from infected flies and used to challenge mice for the production of antisera. ORF3, ORF4a, ORF4b, and ORF4c were individually cloned and expressed in E. coli; resultant recombinant proteins purified and were used to make monospecific antisera. Antisera were evaluated via Western blot against whole virus particles and Nora virus infected fly lysates. Viral purification yielded two particle types with densities of ~1.31 g/mL (empty particles) and ~1.33 g/mL (complete virions). Comparison of purified virus polypeptide composition to Nora virus infected D. melanogaster lysate showed the number of proteins in infected cell lysates is less than purified virus. Our results suggest the virion is composed of 6 polypeptides, VP3, VP4a, two forms of VP4b, and two forms of VP4c. This polypeptide composition is similar to other small RNA insect viruses. PMID:27298753

  12. Simultaneous Detection and Identification of Candida, Aspergillus, and Cryptococcus Species by Reverse Line Blot Hybridization

    PubMed Central

    Playford, E. Geoffrey; Kong, Fanrong; Sun, Ying; Wang, Hui; Halliday, Catriona; Sorrell, Tania C.

    2006-01-01

    We report on a reverse line blot (RLB) assay, utilizing fungal species-specific oligonucleotide probes to hybridize with internal transcribed spacer 2 region sequences amplified using a nested panfungal PCR. Reference and clinical strains of 16 Candida species (116 strains), Cryptococcus neoformans (five strains of Cryptococcus neoformans var. neoformans, five strains of Cryptococcus neoformans var. grubii, and six strains of Cryptococcus gatti), and five Aspergillus species (68 strains) were all correctly identified by the RLB assay. Additional fungal species (16 species and 26 strains) not represented on the assay did not exhibit cross-hybridization with the oligonucleotide probes. In simulated clinical specimens, the sensitivity of the assay for Candida spp. and Aspergillus spp. was 100.5 cells/ml and 102 conidia/ml, respectively. This assay allows sensitive and specific simultaneous detection and identification of a broad range of fungal pathogens. PMID:16517870

  13. A Laboratory Exercise Illustrating the Sensitivity and Specificity of Western Blot Analysis

    ERIC Educational Resources Information Center

    Chang, Ming-Mei; Lovett, Janice

    2011-01-01

    Western blot analysis, commonly known as "Western blotting," is a standard tool in every laboratory where proteins are analyzed. It involves the separation of polypeptides in polyacrylamide gels followed by the electrophoretic transfer of the separated polypeptides onto a nitrocellulose or polyvinylidene fluoride membrane. A replica of the…

  14. Preparation of Cell Lysate from Mouse Oocytes for Western Blotting Analysis.

    PubMed

    Marangos, Petros

    2016-01-01

    Western Blotting has been used extensively for the identification of the protein factors that regulate mammalian oocyte meiosis. However, the limitations in collecting sufficient numbers of oocytes can hinder the efficiency of the technique. Here we provide a detailed protocol for the accurate preparation of mouse oocyte samples for Western Blotting analysis. PMID:27557583

  15. The necessity of and strategies for improving confidence in the accuracy of western blots

    PubMed Central

    Ghosh, Rajeshwary; Gilda, Jennifer E.; Gomes, Aldrin V.

    2016-01-01

    Summary Western blotting is one of the most commonly used laboratory techniques for identifying proteins and semi-quantifying protein amounts, however, several recent findings suggest that western blots may not be as reliable as previously assumed. This is not surprising since many labs are unaware of the limitations of western blotting. In this manuscript we review essential strategies for improving confidence in the accuracy of western blots. These strategies include selecting the best normalization standard, proper sample preparation, determining the linear range for antibodies and protein stains relevant to the sample of interest, confirming the quality of the primary antibody, preventing signal saturation and accurately quantifying the signal intensity of the target protein. Although western blotting is a powerful and indispensable scientific technique that can be used to accurately quantify relative protein levels, it is necessary that proper experimental techniques and strategies are employed. PMID:25059473

  16. The necessity of and strategies for improving confidence in the accuracy of western blots.

    PubMed

    Ghosh, Rajeshwary; Gilda, Jennifer E; Gomes, Aldrin V

    2014-10-01

    Western blotting is one of the most commonly used laboratory techniques for identifying proteins and semi-quantifying protein amounts; however, several recent findings suggest that western blots may not be as reliable as previously assumed. This is not surprising since many labs are unaware of the limitations of western blotting. In this manuscript, we review essential strategies for improving confidence in the accuracy of western blots. These strategies include selecting the best normalization standard, proper sample preparation, determining the linear range for antibodies and protein stains relevant to the sample of interest, confirming the quality of the primary antibody, preventing signal saturation and accurately quantifying the signal intensity of the target protein. Although western blotting is a powerful and indispensable scientific technique that can be used to accurately quantify relative protein levels, it is necessary that proper experimental techniques and strategies are employed. PMID:25059473

  17. Molecular combing compared to Southern blot for measuring D4Z4 contractions in FSHD.

    PubMed

    Vasale, Jessica; Boyar, Fatih; Jocson, Michael; Sulcova, Vladimira; Chan, Patricia; Liaquat, Khalida; Hoffman, Carol; Meservey, Marc; Chang, Isabell; Tsao, David; Hensley, Kerri; Liu, Yan; Owen, Renius; Braastad, Corey; Sun, Weimin; Walrafen, Pierre; Komatsu, Jun; Wang, Jia-Chi; Bensimon, Aaron; Anguiano, Arturo; Jaremko, Malgorzata; Wang, Zhenyuan; Batish, Sat; Strom, Charles; Higgins, Joseph

    2015-12-01

    We compare molecular combing to Southern blot in the analysis of the facioscapulohumeral muscular dystrophy type 1 locus (FSHD1) on chromosome 4q35-qter (chr 4q) in genomic DNA specimens sent to a clinical laboratory for FSHD testing. A de-identified set of 87 genomic DNA specimens determined by Southern blot as normal (n = 71), abnormal with D4Z4 macrosatellite repeat array contractions (n = 7), indeterminate (n = 6), borderline (n = 2), or mosaic (n = 1) was independently re-analyzed by molecular combing in a blinded fashion. The molecular combing results were identical to the Southern blot results in 75 (86%) of cases. All contractions (n = 7) and mosaics (n = 1) detected by Southern blot were confirmed by molecular combing. Of the 71 samples with normal Southern blot results, 67 (94%) had concordant molecular combing results. The four discrepancies were either mosaic (n = 2), rearranged (n = 1), or borderline by molecular combing (n = 1). All indeterminate Southern blot results (n = 6) were resolved by molecular combing as either normal (n = 4), borderline (n = 1), or rearranged (n = 1). The two borderline Southern blot results showed a D4Z4 contraction on the chr 4qA allele and a normal result by molecular combing. Molecular combing overcomes a number of technical limitations of Southern blot by providing direct visualization of D4Z4 macrosatellite repeat arrays on specific chr 4q and chr 10q alleles and more precise D4Z4 repeat sizing. This study suggests that molecular combing has superior analytical validity compared to Southern blot for determining D4Z4 contraction size, detecting mosaicism, and resolving borderline and indeterminate Southern blot results. Further studies are needed to establish the clinical validity and diagnostic accuracy of these findings in FSHD. PMID:26420234

  18. Analysis of common mitochondrial DNA mutations by allele-specific oligonucleotide and Southern blot hybridization.

    PubMed

    Tang, Sha; Halberg, Michelle C; Floyd, Kristen C; Wang, Jing

    2012-01-01

    Mitochondrial disorders are clinically and genetically heterogeneous. There are a set of recurrent point mutations in the mitochondrial DNA (mtDNA) that are responsible for common mitochondrial diseases, including MELAS (mitochondrial encephalopathy, lactic acidosis, stroke-like episodes), MERRF (myoclonic epilepsy and ragged red fibers), LHON (Leber's hereditary optic neuropathy), NARP (neuropathy, ataxia, retinitis pigmentosa), and Leigh syndrome. Most of the pathogenic mtDNA point mutations are present in the heteroplasmic state, meaning that the wild-type and mutant-containing mtDNA molecules are coexisting. Clinical heterogeneity may be due to the degree of mutant load (heteroplasmy) and distribution of heteroplasmic mutations in affected tissues. Additionally, Kearns-Sayre syndrome and Pearson syndrome are caused by large mtDNA deletions. In this chapter, we describe a multiplex PCR/allele-specific oligonucleotide (ASO) hybridization method for the screening of 13 common point mutations. This method allows the detection of low percentage of mutant heteroplasmy. In addition, a nonradioactive Southern blot hybridization protocol for the analysis of mtDNA large deletions is also described. PMID:22215554

  19. OVINE PROGRESSIVE PNEUMONIA VIRUS CAPSID IS B-CELL IMMUNODOMINANT USING WESTERN BLOT ANALYSIS: A COMPARISON OF SENSITIVITY BETWEEN WESTERN BLOT ANALYSIS AND IMMUNOPRECIPITATION

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A western blot assay (WB) was developed and analyzed against the comparable standard, immunoprecipitation of 35[S] methionine/cysteine-labeled ovine progressive pneumonia virus (OPPV) proteins (IP), for its ability to detect anti-OPPV antibodies using endpoint titers. WB is 12-fold more sensitive i...

  20. A Proteomics Approach to the Protein Normalization Problem: Selection of Unvarying Proteins for MS-Based Proteomics and Western Blotting.

    PubMed

    Wiśniewski, Jacek R; Mann, Matthias

    2016-07-01

    Proteomics and other protein-based analysis methods such as Western blotting all face the challenge of discriminating changes in the levels of proteins of interest from inadvertent changes in the amount loaded for analysis. Mass-spectrometry-based proteomics can now estimate the relative and absolute amounts of thousands of proteins across diverse biological systems. We reasoned that this new technology could prove useful for selection of very stably expressed proteins that could serve as better loading controls than those traditionally employed. Large-scale proteomic analyses of SDS lysates of cultured cells and tissues revealed deglycase DJ-1 as the protein with the lowest variability in abundance among different cell types in human, mouse, and amphibian cells. The protein constitutes 0.069 ± 0.017% of total cellular protein and occurs at a specific concentration of 34.6 ± 8.7 pmol/mg of total protein. Since DJ-1 is ubiquitous and therefore easily detectable with several peptides, it can be helpful in normalization of proteomic data sets. In addition, DJ-1 appears to be an advantageous loading control for Western blot that is superior to those used commonly used, allowing comparisons between tissues and cells originating from evolutionarily distant vertebrate species. Notably, this is not possible by the detection and quantitation of housekeeping proteins, which are often used in the Western blot technique. The approach introduced here can be applied to select the most appropriate loading controls for MS-based proteomics or Western blotting in any biological system. PMID:27297043

  1. Easy and Fast Western Blotting by Thin-Film Direct Coating with Suction.

    PubMed

    Liu, Chao-Yuan; Lu, De-Chao; Jiang, Yi-Wei; Yen, Yi-Kuang; Chang, Shih-Chung; Wang, An-Bang

    2016-06-21

    Thin-film direct coating (TDC) has been successfully used in Western blotting (WB). In this study, the advanced technique of TDC with suction (TDCS) was developed to reduce the consumption amount of antibody by a factor of up to 10(4) in comparison with the amount consumed by the conventional WB using the capillary tube without any need of special micromachining processes. The operation time for completely finishing a high-quality WB can be reduced from 3 h in conventional WB to about 5 min or even less by TDCS. In addition, the signal-to-noise ratio of the immunoblotting by TDCS can be markedly increased. TDCS WB showed a high linearity within a 6-log2 dynamic range for detecting 90-6000 ng of purified recombinant glutathione-S-transferase (GST) proteins and could particularly detect extrinsic GST proteins added in crude Escherichia coli or 293T cell lysates. Moreover, a protein mixture containing bovine serum albumin, GST, and ubiquitin could be specifically probed in parallel with their corresponding antibodies through multichannel TDCS WB. This simple and innovative TDCS WB offers various potential applications in simultaneously finishing multiple antibody-antigen screenings in a fast and single experiment. PMID:27254752

  2. Multiplexed microfluidic blotting of proteins and nucleic acids by parallel, serpentine microchannels.

    PubMed

    He, Sha; Zhang, Yi; Wang, Pei; Xu, Xingzhi; Zhu, Kui; Pan, Wenying; Liu, Wenwen; Cai, Kaiyong; Sun, Jiashu; Zhang, Wei; Jiang, Xingyu

    2015-01-01

    This work develops a high-throughput, high-efficiency and straightforward microfluidic blotting method for analyzing proteins and nucleic acids. Sample solutions containing antibodies (for protein detection) or hybridization probes (for nucleic acid detection) are introduced into the parallel, serpentine microchannels to specifically recognize the immobilized targets on the substrate, achieving the identification of multiple targets in multiple samples simultaneously. The loading control, molecular weight markers, and antigen/antibody titration are designed and integrated into the microfluidic chip, thus allowing for the quantification of proteins and nucleic acids. Importantly, we could easily distinguish the adjacent blotting bands inside parallel microchannels, which may be difficult to achieve in conventional blotting. The small dimensions of microfluidic channels also help to reduce the amount of probing molecules and to accelerate the biochemical reaction. Our microfluidic blotting could bypass the steps of blocking and washing, further reducing the operation time and complexity. PMID:25342223

  3. Detection of DNA sequence polymorphisms in human genomic DNA by using denaturing gradient gel blots

    SciTech Connect

    Gray, M.R. )

    1992-02-01

    Denaturing gradient gel electrophoresis can detect sequence differences outside restriction-enzyme recognition sites. DNA sequence polymorphisms can be detected as restriction-fragment melting polymorphisms (RFMPs) in genomic DNA by using blots made from denaturing gradient gels. In contrast to the use of Southern blots to find sequence differences, denaturing gradient gel blots can detect differences almost anywhere, not just at 4-6-bp restriction-enzyme recognition sites. Human genomic DNA was digested with one of several randomly selected 4-bp recognition-site restriction enzymes, electrophoresed in denaturing gradient gels, and transferred to nylon membranes. The blots were hydridized with radioactive probes prepared from the factor VIII, type II collagen, insulin receptor, [beta][sub 2]-adrenergic receptor, and 21-hydroxylase genes; in unrelated individuals, several RFM's were found in fragments from every locus tested. No restriction map or sequence information was used to detect RFMP's.

  4. SOLID-PHASE ASSAY FOR THE PHOSPHORYLATION OF PROTEINS BLOTTED ON NITROCELLULOSE MEMBRANE FILTERS

    EPA Science Inventory

    A new procedure for the phosphorylation and assay of phosphoproteins is described. Proteins are solubilized from tissue samples, separated by polyacrylamide gel electrophoresis, transferred onto nitrocellulose membrane filters and the blotted polypeptides are phosphorylated with ...

  5. Identification of immunodiagnostic antigens for cerebrospinal filariasis in horses by western blot analysis

    PubMed Central

    TAKESUE, Masataka; OSAKA, Yuki; MURANAKA, Masanori; KATAYAMA, Yoshinari; IKADAI, Hiromi

    2016-01-01

    ABSTRACT In the present study, the serum and cerebrospinal fluid of horses diagnosed with Setaria digitata cerebrospinal filariasis were analyzed by western blot. The results revealed S. digitata protein bands measuring 65, 34, 22, and 18 kDa in molecular weight. In particular, the 18 kDa band is a possible candidate for clinical immunodiagnosis on the basis of western blot findings. PMID:27073332

  6. Accuracy of Reverse Dot-Blot PCR in Detection of Different β-Globin Gene Mutations.

    PubMed

    El-Fadaly, N; Abd-Elhameed, A; Abd-Elbar, E; El-Shanshory, M

    2016-06-01

    Prevention programs for β-thalassemia based on molecular diagnosis of heterozygous carriers and/or patients require the use of reliable mutation screening methods. The aim of this study was to compare between direct DNA sequencing, and reverse dot-blot PCR in detection of different β-globin gene mutations in Egyptian children with β-thalassemia. Forty children with β-thalassemia were subjected to mutation analysis, performed by both direct DNA sequencing and β-globin Strip Assay MED™ (based on reverse dot-blot PCR). The most frequent mutant alleles detected by reverse dot-blot PCR were; IVSI-110 G>A (31.25 %), IVS I-6 T > C (21.25 %), and IVS I-1 G>A (20 %). Relatively less frequent mutant alleles detected by reverse dot-blot PCR were "IVSII-1 G>A (5 %), IVSII-745 C>G (5 %), IVSII-848 C>A (2.5 %), IVSI-5 G>C (2.5 %), -87 C>G(2.5 %), and cd39 C>T (2.5 %)", While the genotypes of three patients (6 alleles 7.5 %) were not detected by reverse dot-blot PCR. Mutant alleles detected by direct DNA sequencing were the same as reverse dot-blot PCR method except it revealed the genotypes of 3 undetected patients (one patient was homozygous IVSI-110 G>A, and two patients were homozygous IVS I-1 G>A. Sensitivity of the reverse dot-blot PCR was 92.5 % when compared to direct DNA sequencing for detecting β-thalassemia mutations. Our results therefore suggest that, direct DNA sequencing may be preferred over reverse dot-blot PCR in critical diagnostic situations like genetic counseling for prenatal diagnosis. PMID:27065589

  7. Telomere length measurement by a novel Luminex-based assay: a blinded comparison to Southern blot.

    PubMed

    Pierce, Brandon L; Jasmine, Farzana; Roy, Shantanu; Zhang, Chenan; Aviv, Abraham; Hunt, Steven C; Ahsan, Habibul; Kibriya, Muhammad G

    2016-01-01

    Telomere length (TL) is a potential biomarker of aging and age-related disease risk. We recently published a novel Luminex-based method for high-throughput, low-cost TL measurement. Here we describe a blinded comparison of the Luminex method to Southern blot, the most precise TL measurement method. Luminex and Southern blot measurements for the same 50 DNA samples were taken in two independent laboratories; each sample was measured twice, several months apart. The inter-assay CV for Luminex ranged from 5.5 to 9.1 (depending on CV estimation method), and Southern blot CV from 1.0 to 1.7. Both measures were inversely associated with age. The correlation between the repeated measurements was 0.66 for Luminex and 0.97 for Southern blot. The correlation between Southern blot and Luminex was 0.65 in round 1 and 0.75 in round 2, and the relationship showed no evidence of non-linearity. Our results demonstrate that the Luminex assay is a valid and reproducible method for high-throughput TL measurement. The Luminex assay involves no DNA amplification, which may make Luminex an attractive alternative to PCR-based TL measurement. PMID:27186324

  8. Telomere length measurement by a novel Luminex-based assay: a blinded comparison to Southern blot

    PubMed Central

    Pierce, Brandon L; Jasmine, Farzana; Roy, Shantanu; Zhang, Chenan; Aviv, Abraham; Hunt, Steven C; Ahsan, Habibul; Kibriya, Muhammad G

    2016-01-01

    Telomere length (TL) is a potential biomarker of aging and age-related disease risk. We recently published a novel Luminex-based method for high-throughput, low-cost TL measurement. Here we describe a blinded comparison of the Luminex method to Southern blot, the most precise TL measurement method. Luminex and Southern blot measurements for the same 50 DNA samples were taken in two independent laboratories; each sample was measured twice, several months apart. The inter-assay CV for Luminex ranged from 5.5 to 9.1 (depending on CV estimation method), and Southern blot CV from 1.0 to 1.7. Both measures were inversely associated with age. The correlation between the repeated measurements was 0.66 for Luminex and 0.97 for Southern blot. The correlation between Southern blot and Luminex was 0.65 in round 1 and 0.75 in round 2, and the relationship showed no evidence of non-linearity. Our results demonstrate that the Luminex assay is a valid and reproducible method for high-throughput TL measurement. The Luminex assay involves no DNA amplification, which may make Luminex an attractive alternative to PCR-based TL measurement. PMID:27186324

  9. Blot-MS of Carbonylated Proteins: A Tool to Identify Oxidized Proteins.

    PubMed

    Ferreira, Rita; Domingues, Pedro; Amado, Francisco; Vitorino, Rui

    2016-01-01

    The efficiency of proteostasis regulation declines during aging and the failure of protein homeostasis is common in age-related diseases. Protein oxidation is a major contributor to the loss of proteome homeostasis, also called "proteostasis," precluding protein misfolding and aggregation. So, the identification of the molecular pathways impaired by protein oxidation will increase the understanding of proteostasis and the pathophysiological conditions related to the loss of proteostasis. Sample derivatization with dinitrophenyl hydrazine and western blot immunoassay detection of carbonylated proteins (commonly known as Oxyblot™) coupled to mass spectrometry (blot-MS) is an attractive methodological approach to identify proteins that are more prone to carbonylation, a typical oxidative modification of amino acid residues. The integration of blot-MS data of carbonylated proteins with bioinformatics tools allows the identification of the biological processes more affected by protein oxidation and that, eventually, result in the loss of proteostasis.In this chapter, we describe a blot-MS methodology to identify the proteins more prone to oxidation in biological samples, as cell and tissue extracts, and biofluids. Analysis of mitochondria isolated from cardiac tissue is provided as an example. Bioinformatic strategy to deal with data retrieved from blot-MS experiments are proposed for the identification of relevant biological processes modulated by oxidative stress stimuli. PMID:27613049

  10. HTLV-I/II seroindeterminate Western blot reactivity in a cohort of patients with neurological disease.

    PubMed

    Soldan, S S; Graf, M D; Waziri, A; Flerlage, A N; Robinson, S M; Kawanishi, T; Leist, T P; Lehky, T J; Levin, M C; Jacobson, S

    1999-09-01

    The human T-cell lymphotropic virus type I (HTLV-I) is associated with a chronic, progressive neurological disease known as HTLV-I-associated myelopathy/tropical spastic paraparesis. Screening for HTLV-I involves the detection of virus-specific serum antibodies by EIA and confirmation by Western blot. HTLV-I/II seroindeterminate Western blot patterns have been described worldwide. However, the significance of this blot pattern is unclear. We identified 8 patients with neurological disease and an HTLV-I/II seroindeterminate Western blot pattern, none of whom demonstrated increased spontaneous proliferation and HTLV-I-specific cytotoxic T lymphocyte activity. However, HTLV-I tax sequence was amplified from the peripheral blood lymphocytes of 4 of them. These data suggest that patients with chronic progressive neurological disease and HTLV-I/II Western blot seroindeterminate reactivity may harbor either defective HTLV-I, novel retrovirus with partial homology to HTLV-I, or HTLV-I in low copy number. PMID:10438355

  11. Protein analysis through Western blot of cells excised individually from human brain and muscle tissue

    PubMed Central

    Koob, A.O.; Bruns, L.; Prassler, C.; Masliah, E.; Klopstock, T.; Bender, A.

    2016-01-01

    Comparing protein levels from single cells in tissue has not been achieved through Western blot. Laser capture microdissection allows for the ability to excise single cells from sectioned tissue and compile an aggregate of cells in lysis buffer. In this study we analyzed proteins from cells excised individually from brain and muscle tissue through Western blot. After we excised individual neurons from the substantia nigra of the brain, the accumulated surface area of the individual cells was 120,000, 24,000, 360,000, 480,000, 600,000 μm2. We used an optimized Western blot protocol to probe for tyrosine hydroxylase in this cell pool. We also took 360,000 μm2 of astrocytes (1700 cells) and analyzed the specificity of the method. In muscle we were able to analyze the proteins of the five complexes of the electron transport chain through Western blot from 200 human cells. With this method, we demonstrate the ability to compare cell-specific protein levels in the brain and muscle and describe for the first time how to visualize proteins through Western blot from cells captured individually. PMID:22402104

  12. Western blotting using in-gel protein labeling as a normalization control: stain-free technology.

    PubMed

    Gilda, Jennifer E; Gomes, Aldrin V

    2015-01-01

    Western blotting is a commonly used laboratory technique for semi-quantifying protein amounts. It is important when quantifying protein expression to account for differences in the amount of total protein loaded onto the gel using a loading control. Common loading controls include housekeeping proteins, such as β-actin or GAPDH, quantified by Western blot, or total protein, quantified using a stain such as Coomassie Brilliant Blue or Ponceau S. A more recently developed method for total protein quantification utilizes stain-free technology, which has a linear dynamic detection range and allows for protein detection on both gels and membranes. Here, we describe the theory and use of stain-free gels for total protein quantification and normalization of Western blots. PMID:25820735

  13. Total protein or high-abundance protein: Which offers the best loading control for Western blotting?

    PubMed

    Thacker, Jonathan S; Yeung, Derrick H; Staines, W Richard; Mielke, John G

    2016-03-01

    Western blotting routinely involves a control for variability in the amount of protein across immunoblot lanes. Normalizing a target signal to one found for an abundantly expressed protein is widely regarded as a reliable loading control; however, this approach is being increasingly questioned. As a result, we compared blotting for two high-abundance proteins (actin and glyceraldehyde 3-phosphate dehydrogenase [GAPDH]) and two total protein membrane staining methods (Ponceau and Coomassie Brilliant Blue) to determine the best control for loading variability. We found that Ponceau staining optimally balanced accuracy and precision, and we suggest that this approach be considered as an alternative to normalizing with a high-abundance protein. PMID:26706797

  14. Ferulic acid enhances IgE binding to peanut allergens in western blots.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Phenolic compounds at high concentrations are known to form insoluble complexes with proteins. We hypothesized that this complex formation could interfere with Western blot and ELISA assays for peanut allergens. To verify this, three simple phenolic compounds (ferulic, caffeic, and chlorogenic acids...

  15. Ferulic acid enhances IgE binding to peanut allergens in western blots.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Because phenolic compounds can precipitate or complex with proteins, we postulated that interactions of phenolics with IgE antibodies help enhance IgE binding to peanut allergens in Western blots. Three different phenolics, such as, ferulic, caffeic and chlorogenic acids were examined. Each was mixe...

  16. Line scanning system for direct digital chemiluminescence imaging of DNA sequencing blots

    SciTech Connect

    Karger, A.E.; Weiss, R.; Gesteland, R.F. Eccles Inst. of Human Genetics, Salt Lake City, UT )

    1993-07-01

    A cryogenically cooled charge-coupled device (CCD) camera equipped with an area CCD array is used in a line scanning system for low-light-level imaging of chemiluminescent DNA sequencing blots. Operating the CCD camera in time-delayed integration (TDI) mode results in continuous data acquisition independent of the length of the CCD array. Scanning is possible with a resolution of 1.4 line pairs/mm at the 50% level of the modulation transfer function. High-sensitivity, low-light-level scanning of chemiluminescent direct-transfer electrophoresis (DTE) DNA sequencing blots is shown. The detection of DNA fragments on the blot involves DNA-DNA hybridization with oligonucleotide-alkaline phosphatase conjugate and 1,2-dioxetane-based chemiluminescence. The width of the scan allows the recording of up to four sequencing reactions (16 lanes) on one scan. The scan speed of 52 cm/h used for the sequencing blots corresponds to a data acquisition rate of 384 pixels/s. The chemiluminescence detection limit on the scanned images is 3.9 [times] 10[sup [minus]18] mol of plasmid DNA. A conditional median filter is described to remove spikes caused by cosmic ray events from the CCD images. 39 refs., 9 refs.

  17. A Guide to Modern Quantitative Fluorescent Western Blotting with Troubleshooting Strategies

    PubMed Central

    Eaton, Samantha L.; Hurtado, Maica Llavero; Oldknow, Karla J.; Graham, Laura C.; Marchant, Thomas W.; Gillingwater, Thomas H.; Farquharson, Colin; Wishart, Thomas M.

    2014-01-01

    The late 1970s saw the first publicly reported use of the western blot, a technique for assessing the presence and relative abundance of specific proteins within complex biological samples. Since then, western blotting methodology has become a common component of the molecular biologists experimental repertoire. A cursory search of PubMed using the term “western blot” suggests that in excess of two hundred and twenty thousand published manuscripts have made use of this technique by the year 2014. Importantly, the last ten years have seen technical imaging advances coupled with the development of sensitive fluorescent labels which have improved sensitivity and yielded even greater ranges of linear detection. The result is a now truly Quantifiable Fluorescence based Western Blot (QFWB) that allows biologists to carry out comparative expression analysis with greater sensitivity and accuracy than ever before. Many “optimized” western blotting methodologies exist and are utilized in different laboratories. These often prove difficult to implement due to the requirement of subtle but undocumented procedural amendments. This protocol provides a comprehensive description of an established and robust QFWB method, complete with troubleshooting strategies. PMID:25490604

  18. 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…

  19. COMPARISONS OF ELISA AND WESTERN BLOT ASSAYS FOR DETECTION OF CRYPTOSPORIDIUM ANTIBODY

    EPA Science Inventory

    A seroprevalence survey was conducted using ELISA and Western blot (WB) assays for antibody to three Cryptosporidium antigens on 380 blood donors in Jackson County, Oregon. The purpose was to determine if either assay could detect serological evidence of an outbreak which occurre...

  20. A Study of Rubisco through Western Blotting and Tissue Printing Techniques

    ERIC Educational Resources Information Center

    Ma, Zhong; Cooper, Cynthia; Kim, Hyun-Joo; Janick-Buckner, Diane

    2009-01-01

    We describe a laboratory exercise developed for a cell biology course for second-year undergraduate biology majors. It was designed to introduce undergraduates to the basic molecular biology techniques of Western blotting and immunodetection coupled with the technique of tissue printing in detecting the presence, relative abundance, and…

  1. Evaluation of immunoglobulin M western blot analysis in the diagnosis of congenital syphilis.

    PubMed Central

    Lewis, L L; Taber, L H; Baughn, R E

    1990-01-01

    Western immunoblots of solubilized Treponema pallidum antigens were reacted with sera and cerebrospinal fluid (CSF) and developed with enzyme-conjugated antibodies to immunoglobulin M (IgM). A blot was considered positive if reactions included bands at the 47-, 17-, and 15.5-kilodalton positions along with a variable pattern at other low-molecular-weight positions. Sera from 23 of 25 symptomatic infants diagnosed with congenital syphilis yielded positive reactions. Of 80 asymptomatic infants considered at risk for developing symptomatic infection, 16 exhibited IgM patterns consistent with those seen in congenital syphilis, although 5 of these 16 gave reactions that were equivocal. To exclude false-positive reactions due to IgM rheumatoid factor, sera were fractionated and the IgM fractions were retested. Only the five initially equivocal sera gave nonreactive blots with the IgM fractions, whereas all others gave more prominent reactions that were qualitatively similar to those seen in serum samples. Sera from 18 normal infants failed to show any IgM reactivity to T. pallidum antigens on Western blots. The IgM Western blot was both more sensitive and more specific than the fluorescent treponemal antibody-absorbed (IgM) test using fractionated serum. Of the 17 CSF samples from infants with symptomatic congenital syphilis, 14 showed IgM reactivity in Western blots, whereas only 12 had a reactive CSF in the Venereal Disease Research Laboratory test. Our results indicate that this technique can be used to identify both symptomatic and asymptomatic infection in infants with T. pallidum, in some cases before standard serologic studies can confirm the diagnosis. Images PMID:2179261

  2. A Slot Blot Immunoassay for Quantitative Detection of Plasmodium falciparum Circumsporozoite Protein in Mosquito Midgut Oocyst

    PubMed Central

    Kumar, Sanjai; Zheng, Hong; Deng, Bingbing; Mahajan, Babita; Grabias, Bryan; Kozakai, Yukiko; Morin, Merribeth J.; Locke, Emily; Birkett, Ashley; Miura, Kazutoyo; Long, Carole

    2014-01-01

    There is still a need for sensitive and reproducible immunoassays for quantitative detection of malarial antigens in preclinical and clinical phases of vaccine development and in epidemiology and surveillance studies, particularly in the vector host. Here we report the results of sensitivity and reproducibility studies for a research-grade, quantitative enhanced chemiluminescent-based slot blot assay (ECL-SB) for detection of both recombinant Plasmodium falciparum circumsporozoite protein (rPfCSP) and native PfCSP from Oocysts (Pf Oocyst) developing in the midguts of Anopheles stephensi mosquitoes. The ECL-SB detects as little as 1.25 pg of rPfCSP (linear range of quantitation 2.5–20 pg; R2 = 0.9505). We also find the earliest detectable expression of native PfCSP in Pf Oocyst by ECL-SB occurs on day 7 post feeding with infected blood meal. The ECL-SB was able to detect approximately as few as 0.5 day 8 Pf Oocysts (linear quantitation range 1–4, R2 = 0.9795) and determined that one Pf Oocyst expressed approximately 2.0 pg (0.5–3 pg) of native PfCSP, suggesting a similar range of detection for recombinant and native forms of Pf CSP. The ECL-SB is highly reproducible; the Coefficient of Variation (CV) for inter-assay variability for rPf CSP and native PfCSP were 1.74% and 1.32%, respectively. The CVs for intra-assay variability performed on three days for rPf CSP were 2.41%, 0.82% and 2% and for native Pf CSP 1.52%, 0.57%, and 1.86%, respectively. In addition, the ECL-SB was comparable to microscopy in determining the P. falciparum prevalence in mosquito populations that distinctly contained either high and low midgut Pf Oocyst burden. In whole mosquito samples, estimations of positivity for P. falciparum in the high and low burden groups were 83.3% and 23.3% by ECL-SB and 85.7% and 27.6% by microscopy. Based on its performance characteristics, ECL-SB could be valuable in vaccine development and to measure the parasite prevalence in mosquitoes and

  3. Northern Nutrition.

    ERIC Educational Resources Information Center

    Northwest Territories Dept. of Education, Yellowknife.

    This guide contains nutrition information and nutrition education strategies aimed at residents of the Canadian Arctic. Section I: (1) defines nutrition terms; (2) describes the sources and functions of essential nutrients; (3) explains Canada's food guide and special considerations for the traditional northern Native diet and for lactose…

  4. Dot Blot Assay for Detection of Antidiacyltrehalose Antibodies in Tuberculous Patients

    PubMed Central

    Vera-Cabrera, Lucio; Rendon, Adrian; Diaz-Rodriguez, Manuel; Handzel, Vera; Laszlo, Adalbert

    1999-01-01

    A simple dot blot test with diacyltrehalose (DAT) as the antigen was developed to detect anti-DAT antibodies in tuberculous patients. To enhance antigen-antibody reaction detection, rabbit serum raised against human immunoglobulins was used prior to incubation with a protein A-colloidal gold complex. With the dot blot system, it was possible to obtain a sensitivity similar to that of enzyme-linked immunosorbent assay (ELISA) and a specificity of 97.14%, versus a specificity of 94.29% by the ELISA. We conclude that this simple and fast assay could be used in places where ELISA equipment is not easy available and that it might also be applicable with other Mycobacterium tuberculosis immunogenic antigens. PMID:10473518

  5. Solid-phase assay for the phosphorylation of proteins blotted on nitrocellulose membrane filters

    SciTech Connect

    Valtorta, F.; Schiebler, W.; Jahn, R.; Ceccarelli, B.; Greengard, P.

    1986-10-01

    A new procedure for the phosphorylation and assay of phosphoproteins is described. Proteins are solubilized from tissue samples, separated by polyacrylamide gel electrophoresis, transferred onto nitrocellulose membrane filters, and the blotted polypeptides are phyosphorylated with the catalytic subunit of cyclic AMP (adenosine 3':5'-monophosphate)-dependent protein kinase. The method was developed for the assay of dephosphosynapsin I, but it has also proven suitable for the phosphorylation of other proteins. The patterns of phosphorylation of tissue samples phosphorylated using the new method are similar to those obtained using the conventional test tube assay. Once phosphorylated, the adsorbed proteins can be digested with proteases and subjected to phosphopeptide mapping. The phosphorylated blotted proteins can also be analyzed by overlay techniques for the immunological detection of polypeptides.

  6. Detection of Diverse and High Molecular Weight Nesprin-1 and Nesprin-2 Isoforms Using Western Blotting.

    PubMed

    Carthew, James; Karakesisoglou, Iakowos

    2016-01-01

    Heavily utilized in cell and molecular biology, western blotting is considered a crucial technique for the detection and quantification of proteins within complex mixtures. In particular, the detection of members of the nesprin (nuclear envelope spectrin repeat protein) family has proven difficult to analyze due to their substantial isoform diversity, molecular weight variation, and the sheer size of both nesprin-1 and nesprin-2 giant protein variants (>800 kDa). Nesprin isoforms contain distinct domain signatures, perform differential cytoskeletal associations, occupy different subcellular compartments, and vary in their tissue expression profiles. This structural and functional variance highlights the need to distinguish between the full range of proteins within the nesprin protein family, allowing for greater understanding of their specific roles in cell biology and disease. Herein, we describe a western blotting protocol modified for the detection of low to high molecular weight (50-1000 kDa) nesprin proteins. PMID:27147045

  7. A Fast and Inexpensive Western Blot Experiment for the Undergraduate Laboratory

    NASA Astrophysics Data System (ADS)

    Farrell, Shawn O.; Farrell, Lynn E.

    1995-08-01

    Western blotting is an important, modern technique for transferring proteins from a gel onto nitrocellulose or other suitable support and then detecting a protein of interest using antibodies. We have developed an experiment and optimized the conditions for the undergraduate laboratory. The experiment can be done quickly using an electrophoretic blotter or more cheaply using passive transfer. This experiment allows the student to learn valuable procedures currently used in biochemistry and other biological sciences.

  8. A slot blot procedure for the measurement of yessotoxins by a functional assay.

    PubMed

    Pierotti, Silvia; Albano, Clara; Milandri, Anna; Callegari, Federica; Poletti, Roberto; Rossini, Gian Paolo

    2007-01-01

    We originally developed a functional assay for the detection of yessotoxins (YTX) based on its capacity to induce dose-dependent changes in cellular levels of two marker proteins, consisting of E-cadherin and an E-cadherin fragment (ECRA100) in epithelial cells. The procedure is time-consuming and we have shortened it by a slot blot format, using antibodies recognizing two different epitopes of E-cadherin (HECD-1 and C20820), thereby discriminating those markers. The best performing membrane under our conditions, in terms of binding capacity and even absorption of proteins, was a positively charged nylon membrane. Treatment of the membrane with 0.5mug of Ab/ml was appropriate for maximal detection of antigens by our slot blot procedure with both HECD-1 and C20820 antibodies. The treatment of cells with YTX, resulting in a relative increase in the cellular levels of ECRA100, led to a dose-dependent increase of the signal detected by Ab HECD-1 without a concomitant increase in the signal detected by Ab C20820 in our slot blot format, and the concentrations of YTX were correlated to both the increase of the signal detected through Ab HECD-1 and to the decrease in the ratio of the signals obtained with the two Abs (C20820 over HECD-1). Upon analyses of extracts from cells treated with shellfish samples, we could detect and quantify YTX in naturally contaminated materials. The slot blot format of our functional assay allows a substantial shortening of its analytical step (about seven hr, as compared to the two working days of the original method), providing YTX measurements that are accurate but show large standard deviations. PMID:17055548

  9. Western Blotting using the Invitrogen NuPage Novex Bis Tris minigels.

    PubMed

    Penna, Aubin; Cahalan, Michael

    2007-01-01

    Western Blotting (or immunoblotting) is a standard laboratory procedure allowing investigators to verify the expression of a protein, determine the relative amount of the protein present in different samples, and analyze the results of co-immunoprecipitation experiments. In this method, a target protein is detected with a specific primary antibody in a given sample of tissue homogenate or extract. Protein separation according to molecular weight is achieved using denaturing SDS-PAGE. After transfer to a membrane, the target protein is probed with a specific primary antibody and detected by chemiluminescence. Since its first description, the western-blotting technique has undergone several improvements, including pre-cast gels and user-friendly equipment. In our laboratory, we have chosen to use the commercially available NuPAGE electrophoresis system from Invitrogen. It is an innovative neutral pH, discontinuous SDS-PAGE, pre-cast mini-gel system. This system presents several advantages over the traditional Laemmli technique including: i) a longer shelf life of the pre-cast gels ranging from 8 months to 1 year; ii) a broad separation range of molecular weights from 1 to 400 kDa depending of the type of gel used; and iii) greater versatility (range of acrylamide percentage, the type of gel, and the ionic composition of the running buffer). The procedure described in this video article utilizes the Bis-Tris discontinuous buffer system with 4-12% Bis-Tris gradient gels and MES running buffer, as an illustration of how to perform a western-blot using the Invitrogen NuPAGE electrophoresis system. In our laboratory, we have obtained good and reproducible results for various biochemical applications using this western-blotting method. PMID:18989435

  10. Detection of Sleeping Beauty transposition in the genome of host cells by non-radioactive Southern blot analysis.

    PubMed

    Aravalli, Rajagopal N; Park, Chang W; Steer, Clifford J

    2016-08-26

    The Sleeping Beauty transposon (SB-Tn) system is being used widely as a DNA vector for the delivery of therapeutic transgenes, as well as a tool for the insertional mutagenesis in animal models. In order to accurately assess the insertional potential and properties related to the integration of SB it is essential to determine the copy number of SB-Tn in the host genome. Recently developed SB100X transposase has demonstrated an integration rate that was much higher than the original SB10 and that of other versions of hyperactive SB transposases, such as HSB3 or HSB17. In this study, we have constructed a series of SB vectors carrying either a DsRed or a human β-globin transgene that was encompassed by cHS4 insulator elements, and containing the SB100X transposase gene outside the SB-Tn unit within the same vector in cis configuration. These SB-Tn constructs were introduced into the K-562 erythroid cell line, and their presence in the genomes of host cells was analyzed by Southern blot analysis using non-radioactive probes. Many copies of SB-Tn insertions were detected in host cells regardless of transgene sequences or the presence of cHS4 insulator elements. Interestingly, the size difference of 2.4 kb between insulated SB and non-insulated controls did not reflect the proportional difference in copy numbers of inserted SB-Tns. We then attempted methylation-sensitive Southern blots to assess the potential influence of cHS4 insulator elements on the epigenetic modification of SB-Tn. Our results indicated that SB100X was able to integrate at multiple sites with the number of SB-Tn copies larger than 6 kb in size. In addition, the non-radioactive Southern blot protocols developed here will be useful to detect integrated SB-Tn copies in any mammalian cell type. PMID:27329815

  11. Restricted specificity of the autoantibody response in Goodpasture's syndrome demonstrated by two-dimensional western blotting.

    PubMed

    Derry, C J; Dunn, M J; Rees, A J; Pusey, C D

    1991-12-01

    The autoantigen in Goodpasture's syndrome is known to be contained within the non-collagenous (NC1) domain of type IV collagen. We have examined the specificity of autoantibodies to glomerular basement membrane (GBM) using the technique of 2-D electrophoresis followed by Western blotting. Protein stains of 2-D gels of collagenase-digested human GBM revealed extensive charge and size heterogeneity. Major components were of mol. wt 24-30 kD and 43-56 kD, corresponding to monomeric and dimeric subunits of NCl. Western blotting of 2-D gels with IgG from patients with anti-GBM disease demonstrated that the most antigenic components migrated as cationic 28-kD monomers (pI 10) and similarly charged dimers, although other components were recognized less strongly. The mobility of the strongly antigenic polypeptides was different to that of the known alpha 1 and alpha 2 chains of type IV collagen. Autoantibodies from all 20 patients studied showed the same pattern of reactivity, regardless of their clinical features (in particular, the presence or absence of pulmonary haemorrhage) or HLA type. A monoclonal antibody (P1) to human GBM bound in a similar pattern, particularly recognizing the cationic components. 2-D gels of affinity-purified GBM from a P1 column showed enrichment of the 28-kD monomers, which were recognized by human autoantibodies on Western blotting. These results demonstrate that the autoimmune response in Goodpasture's syndrome is of restricted specificity, and support the suggestion that the major autoantigenic determinant is present on the novel alpha 3 chain of type IV collagen. PMID:1747953

  12. Fluorescent detection of Southern blots and PCR-based genetic typing tests

    SciTech Connect

    Mansfield, E.S.; Worley, J.M.; Zimmerman, P.A.

    1994-09-01

    The Southern blot is used to study gene organization, to identify disease-causing genomic rearrangements, or for typing RFLP markers in forensic, paternity, or prenatal diagnostic testing. Fluorescence offers a much greater dynamic range and a more linear response than film used in radioactive or chemiluminescent detection of RFLPs. We therefore investigated using the Fluorimager{trademark} 575 (Molecular Dynamics, Inc.) for analyzing Southern blots. Using a single-locus probe to D2S44 (YNH24) (Promega Corp.), we detect as little as 100 ng (0.05 attomole) genomic DNA. The alkaline phosphatase-labeled probe is detected using AttoPhos (JBL Scientific), and the developed membrane is scanned with the Fluorimager. Biotinylated hybridization probes can also be developed using a streptavidin-alkaline phosphatase conjugate and AttoPhos. The instrument scan parameters can be adjusted to prevent overexposure and accompanying loss of resolution in images of blots, gels, or 96-well microplates. We have used these other sample formats in PCR-based genetic typing assays. We use FluorKit DQS (Molecular Dynamics) to accurately quantify PCR template DNA (1-500 ng) in 96-well microplates scanned using the same instrument. Mutation detection assays run include heteroduplex gels (5% polyacrylamide, 2.7 M urea), short tandem repeat (STR) markers, amplified fragment length polymorphisms (AmpFLP), competitive priming PCR, and allele-specific oligotyping. These assays are run using either 1- or 2-color labeling. We detect unlabeled PCR products, such as the AmpFLP marker D1S80 (Perkin-Elmer) by post-staining gels for 10 minutes with SYBR Green 1 (Molecular Probes) and scanning the wet gel. The Fluorimager scans a 20 x 25 cm sample within three minutes, allowing rapid optimization of fluorescent protocols and high sample throughput.

  13. RNA Colony Blot Hybridization Method for Enumeration of Culturable Vibrio cholerae and Vibrio mimicus Bacteria▿

    PubMed Central

    Grim, Christopher J.; Zo, Young-Gun; Hasan, Nur A.; Ali, Afsar; Chowdhury, Wasimul B.; Islam, Atiqul; Rashid, Mohammed H.; Alam, Munirul; Morris, J. Glenn; Huq, Anwar; Colwell, Rita R.

    2009-01-01

    A species-specific RNA colony blot hybridization protocol was developed for enumeration of culturable Vibrio cholerae and Vibrio mimicus bacteria in environmental water samples. Bacterial colonies on selective or nonselective plates were lysed by sodium dodecyl sulfate, and the lysates were immobilized on nylon membranes. A fluorescently labeled oligonucleotide probe targeting a phylogenetic signature sequence of 16S rRNA of V. cholerae and V. mimicus was hybridized to rRNA molecules immobilized on the nylon colony lift blots. The protocol produced strong positive signals for all colonies of the 15 diverse V. cholerae-V. mimicus strains tested, indicating 100% sensitivity of the probe for the targeted species. For visible colonies of 10 nontarget species, the specificity of the probe was calculated to be 90% because of a weak positive signal produced by Grimontia (Vibrio) hollisae, a marine bacterium. When both the sensitivity and specificity of the assay were evaluated using lake water samples amended with a bioluminescent V. cholerae strain, no false-negative or false-positive results were found, indicating 100% sensitivity and specificity for culturable bacterial populations in freshwater samples when G. hollisae was not present. When the protocol was applied to laboratory microcosms containing V. cholerae attached to live copepods, copepods were found to carry approximately 10,000 to 50,000 CFU of V. cholerae per copepod. The protocol was also used to analyze pond water samples collected in an area of cholera endemicity in Bangladesh over a 9-month period. Water samples collected from six ponds demonstrated a peak in abundance of total culturable V. cholerae bacteria 1 to 2 months prior to observed increases in pathogenic V. cholerae and in clinical cases recorded by the area health clinic. The method provides a highly specific and sensitive tool for monitoring the dynamics of V. cholerae in the environment. The RNA blot hybridization protocol can also be

  14. Neurofilament dot blot assays: novel means of assessing axon viability in culture.

    PubMed

    Hares, Kelly; Kemp, Kevin; Gray, Elizabeth; Scolding, Neil; Wilkins, Alastair

    2011-06-15

    Axonal structure and integrity are vital to overall neuronal maintenance and action potential propagation. Neurofilaments (NFs) are one of the main cytoskeletal components of axons and phosphorylation of NF subunits regulates speed of NF transport through axons and determines optimal axonal calibre required for signal propagation. Many previous studies of neuroprotective agents have focussed on neuronal viability in models of neurodegenerative disease, without specifically considering axon function as an indicator of neuronal damage. In this study, we have focused on developing novel assays for determining axon viability by measuring levels of neurofilament phosphorylation in cultured cortical neurons. The nitric oxide donor DETANONOate (NO) was used as an inflammatory insult and glial cell line-derived neurotrophic factor (GDNF) and superoxide dismutase (SOD) were tested as potential axonal protective agents. Using 'dot blot' methodologies, we show a decrease in NF phosphorylation in cortical neurons exposed to NO-mediated cell toxicity and an attenuation of NO-mediated changes in NF phosphorylation associated with GDNF and SOD treatment. These results correlated well with immunocytochemical counts. We propose therefore that the dot blot assay is a novel method for assessing axonal integrity in vitro and may play a useful role in the future for testing the effects of agents on axonal viability, providing a reliable and reproducible screening method for potential therapeutics for neurodegenerative diseases. PMID:21459112

  15. Validation of Endothelin B Receptor Antibodies Reveals Two Distinct Receptor-related Bands on Western Blot

    PubMed Central

    Barr, Travis P.; Kornberg, Daniel; Montmayeur, Jean-Pierre; Long, Melinda; Reichheld, Stephen; Strichartz, Gary R.

    2014-01-01

    Antibodies are important tools for the study of protein expression, but are often used without full validation. In this study, we use Western blots to characterize antibodies targeted to the N- (NT) or C-termini (CT) and the second (IL2) or third intracellular (IL3) loops of the endothelin B receptor (ETB). The IL2-targeted antibody accurately detected endogenous ETB expression in rat brain and cultured rat astrocytes by labeling a 50kD band, the expected weight of full-length ETB. However, this antibody failed to detect transfected ETB in HEK293 cultures. In contrast, the NT-targeted antibody accurately detected endogenous ETB in rat astrocyte cultures and transfected ETB in HEK293 cultures by labeling a 37 kD band, but failed to detect endogenous ETB in rat brain. Bands detected by the CT-targeted or IL3-targeted antibodies were found to be unrelated to ETB. Our findings show that functional ETB receptors can be detected at 50 kD or 37 kD on Western blot, with drastic differences in antibody affinity for these bands. The 37 kD band likely reflects ETB receptor processing, which appears to be dependent on cell type and/or culture condition. PMID:25232999

  16. Evaluating cytoplasmic and nuclear levels of inflammatory cytokines in cancer cells by western blotting.

    PubMed

    Gatla, Himavanth R; Singha, Bipradeb; Persaud, Valerie; Vancurova, Ivana

    2014-01-01

    Increased expression and cellular release of inflammatory cytokines, interleukin-8 (IL-8; CXCL8), and high mobility group box-1 (HMGB1) are associated with increased cell proliferation, angiogenesis, and metastasis during cancer progression. In prostate and ovarian cancer cells, increased levels of IL-8 and HMGB1 correlate with poor prognosis. We have recently shown that proteasome inhibition by bortezomib (BZ) specifically increases IL-8 release from metastatic prostate and ovarian cancer cells. In this chapter, we describe a protocol to analyze the cytoplasmic and nuclear levels of IL-8 and HMGB1 in prostate and ovarian cancer cells by western blotting. IL-8 is localized in the cytoplasm in both cell types, and its protein levels are significantly increased by BZ. In contrast, HMGB1 is localized in the nucleus, and BZ increases its nuclear levels only in ovarian cancer cells. The protocol includes isolation of cytoplasmic and nuclear extracts, followed by SDS electrophoresis and western blotting, and can be easily modified to analyze the cytoplasmic and nuclear cytokine levels in other cell types. PMID:24908314

  17. Immunodiagnosis of Echinococcus Infections: Confirmatory Testing and Species Differentiation by a New Commercial Western Blot

    PubMed Central

    Liance, Martine; Janin, Veronique; Bresson-Hadni, Solange; Vuitton, Dominique-Angele; Houin, Rene; Piarroux, Renaud

    2000-01-01

    The Echinococcus Western Blot IgG (LDBIO Diagnostics, Lyon, France), using a whole larval antigen from Echinococcus multilocularis, was evaluated for serodiagnosis and differentiation between two human parasitic infections of worldwide importance: cystic echinococcosis, due to Echinococcus granulosus, and alveolar echinococcosis, due to E. multilocularis. Fifty and 61 serum samples from patients with cystic and alveolar echinococcosis, respectively, were used for assessing diagnostic sensitivity. The sensitivity of the assay was compared with those of screening tests used for these applications. Sera used for assessing cross-reactivities were from 154 patients with other diseases, either parasitic or not. The assay allowed the detection of serum immunoglobulin G antibodies in 97% of Echinococcus-infected patients. It had a higher sensitivity than screening assays for the detection for each echinococcosis. The assay allowed us to correctly distinguish between E. granulosus- and E. multilocularis-infected patients in 76% of cases. It did not allow us to distinguish active from inactive forms of both echinococcoses. The occurrence of cross-reactivities with neurocysticercosis indicates the necessity for retesting sera with species-specific antigens, for rare patients with neurologic disorders. This study shows the usefulness of the commercially available Echinococcus Western Blot IgG for the serological confirmation of human echinococcosis. PMID:11015390

  18. Visualization of heparin-binding proteins by ligand blotting with /sup 125/I-heparin

    SciTech Connect

    Cardin, A.D.; Witt, K.R.; Jackson, R.L.

    1984-03-01

    A ligand-blotting procedure which allows detection of heparin-binding proteins is described. Crude commercial heparin was fractionated by chromatography on a column of human plasma low-density lipoproteins immobilized to Sepharose CL-4B. Chromatography yielded an unbound and a bound fraction of heparin, designated URH and HRH, respectively. The HRH fraction was reacted with the N-hydroxysuccinimidyl ester of 3-(p-hydroxyphenyl)propionic acid and then labeled with /sup 125/I. Proteins were separated by 3-20% pore-gradient gel electrophoresis, transferred to nitrocellulose, and then assayed for their ability to bind /sup 125/I-labeled HRH. Human plasma apolipoproteins B-100, B-48, and E of chylomicrons, very low-density lipoproteins, and low-density lipoproteins bound the /sup 125/I-labeled HRH; the radiolabeled haparin did not bind to serum albumin, ferritin, catalase, and lactate dehydrogenase. The ligand-blotting procedure should facilitate the purification of heparin-binding domains from these proteins and, moreover may be applicable to the investigation of heparin-protein interactions in general. 15 references.

  19. Analysis of sperm antigens by sodium dodecyl sulfate gel/protein blot radioimmunobinding method

    SciTech Connect

    Lee, C.Y.G.; Huang, Y.S.; Hu, P.C.; Gomel, V.; Menge, A.C.

    1982-06-01

    A radioimmunobinding method based on the blotting of renatured proteins from sodium dodecyl sulfate gels on to nitrocellulose filter papers was developed to analyze the sperm antigens that elicit serum anti-sperm antibodies. In rabbits, serum anti-sperm antibodies were raised by immunization with homologous epididymal spermatozoa mixed with complete Freund's adjuvant. The raised antisera from either male or female rabbits were shown to react with three major sperm protein bands on sodium dodecyl sulfate gels with the corresponding molecular weights of about 70,000 +/- 5000, 14,000, and 13,000, respectively. In humans, the monoclonal antibodies against human sperm were raised by a hybridoma technique. Out of six independent hybrid cell lines that were generated, three of them were shown to secrete immunoglobulins that react with the same two protein bands on sodium dodecyl sulfate gels, which have the approximate molecular weight of 10,000. The same procedure was also used to analyze human serum samples that were shown to contain anti-sperm antibodies by the known techniques. Unique sperm antigens that elicit anti-sperm antibodies in humans were identified and correlated. The results of this study suggest that sodium dodecyl sulfate gel/protein blot radioimmunobinding method may be a sensitive and useful tool for the study of sperm antigens that elicit autoimmune responses and their association with human infertility.

  20. Western thymomas lack Epstein-Barr virus by Southern blotting analysis and by polymerase chain reaction.

    PubMed Central

    Inghirami, G.; Chilosi, M.; Knowles, D. M.

    1990-01-01

    The authors investigated 16 western thymomas, 9 from the United States and 7 from Europe, for the presence of Epstein-Barr virus (EBV) DNA sequences by both Southern blot hybridization analysis and polymerase chain reaction using EBV-specific DNA probes that detect the long internal repeat and terminal repeat regions and the EBNA-1 gene. None of the 16 thymomas contained evidence of the EBV genome, even though we could detect EBV by Southern blotting when EBV DNA represents less than or equal to 1% of the total DNA and by polymerase chain reaction when a single EBV-positive cell is present among 10(5) EBV-negative cells. These results fail to demonstrate EBV genome in western thymomas and stand in contrast to those of McGuire et al (Am J Pathol 1988, 131:385) who previously reported that the EBV genome is present in thymomas occurring in southern Chinese patients. Therefore EBV does not appear to be implicated in the pathogenesis of all thymomas. The presence of EBV in eastern thymomas, regions where EBV is endemic may be due to epidemiologic factors and/or genetic predispositions. Images Figure 1 Figure 2 Figure 3 Figure 4 PMID:2162629

  1. Comparative analysis of human papillomavirus detection by dot blot hybridisation and non-isotopic in situ hybridisation.

    PubMed Central

    Troncone, G; Anderson, S M; Herrington, C S; de Angelis, M L; Noell, H; Chimera, J A; O'D McGee, J

    1992-01-01

    AIMS: To determine the relative diagnostic performance of non-isotopic in situ hybridisation (NISH) and a dot-blot assay for detecting human papillomavirus (HPV) on exfoliated cervical cells; and to correlate the results with cytopathological assessment. METHODS: Cervical smears and cytological samples were obtained from 122 patients during the same clinical examination and the presence of HPV sequences determined by NISH and dot-blot analysis, respectively. RESULTS: Dot-blot analysis gave an autoradiographic signal in 15 of 121 (12.4%) cases, while NISH detected viral genomes in 38 of 114 (33.3%) cases. Even in the presence of koilocytosis, where vegetative replication of the virus occurs, NISH was positive in over twice as many cases as dot-blot analysis (NISH 90%, dot-blot 40%), while in smears within normal cytological limits, where the viral copy number is likely to be considerably lower, the differences were more striking (NISH 31%, dot-blot 5%). CONCLUSIONS: These data show that NISH on cytological smears is more sensitive than a standardised dot-blot hybridisation assay for detecting HPV infection in cytological material and is therefore a more appropriate screening tool. Images PMID:1331197

  2. Parity among the randomly amplified polymorphic DNA method, multilocus enzyme electrophoresis, and Southern blot hybridization with the moderately repetitive DNA probe Ca3 for fingerprinting Candida albicans.

    PubMed Central

    Pujol, C; Joly, S; Lockhart, S R; Noel, S; Tibayrenc, M; Soll, D R

    1997-01-01

    Randomly amplified polymorphic DNA (RAPD) analysis, multilocus enzyme electrophoresis (MLEE), and Southern blot hybridization with moderately repetitive DNA probes have emerged as effective fingerprinting methods for the infectious fungus Candida albicans. The three methods have been compared for their capacities to identify identical or highly related isolates, to cluster weakly related isolates, to discriminate between unrelated isolates, and to assess microevolution within a strain. By computing similarity coefficients between 29 isolates from three cities within the continental United States, strong concordance of the results is demonstrated for RAPD analysis, MLEE, and Southern blot hybridization with the moderately repetitive probe Ca3, and weaker concordance of the results is demonstrated for these three fingerprinting methods and Southern blot hybridization with the moderately repetitive probe CARE2. All methods were also demonstrated to be able to resolve microevolution within a strain, with the Ca3 probe exhibiting the greatest resolving power. The strong correlations demonstrated between polymorphic markers assessed by the four independent fingerprinting methods and the nonrandom association between loci demonstrated by RAPD analysis and MLEE provide evidence for strong linkage disequilibrium and a clonal population structure for C. albicans. In addition, a synapomorphic allele, Pep-3A, was found to be present in all members of one of the three clusters discriminated by RAPD analysis, MLEE, and Ca3 fingerprinting, supporting the concordance of the clustering capacities of the three methods, the robustness of the clusters, and the clonal nature of the clusters. PMID:9276415

  3. Western Blot Detection of Human Anti-Chikungunya Virus Antibody with Recombinant Envelope 2 Protein.

    PubMed

    Yang, Zhaoshou; Lee, Jihoo; Ahn, Hye-Jin; Chong, Chom-Kyu; Dias, Ronaldo F; Nam, Ho-Woo

    2016-04-01

    Chikungunya virus (CHIKV), a tropical pathogen, has re-emerged and has massive outbreaks abruptly all over the world. Containing many dominant epitopes, the envelope E2 protein of CHIKV has been explored for the vaccination or diagnosis. In the present study, the antigenicity of a recombinant expressed intrinsically disorder domain (IUD) of E2 was tested for the detection of the antibody against CHIKV through western blot method. The gene of the IUD of E2 was inserted into 2 different vectors and expressed as recombinant GST-E2 and recombinant MBP-E2 fusion protein, respectively. Two kinds of fusion proteins were tested with 30 CHIKV patient sera and 30 normal sera, respectively. Both proteins were detected by 25 patients sera (83.3%) and 1 normal serum (3.3%). This test showed a relatively high sensitivity and very high specificity of the recombinant E2 proteins to be used as diagnostic antigens against CHIKV infection. PMID:27180586

  4. Western Blot Detection of Human Anti-Chikungunya Virus Antibody with Recombinant Envelope 2 Protein

    PubMed Central

    Yang, Zhaoshou; Lee, Jihoo; Ahn, Hye-Jin; Chong, Chom-Kyu; Dias, Ronaldo F.; Nam, Ho-Woo

    2016-01-01

    Chikungunya virus (CHIKV), a tropical pathogen, has re-emerged and has massive outbreaks abruptly all over the world. Containing many dominant epitopes, the envelope E2 protein of CHIKV has been explored for the vaccination or diagnosis. In the present study, the antigenicity of a recombinant expressed intrinsically disorder domain (IUD) of E2 was tested for the detection of the antibody against CHIKV through western blot method. The gene of the IUD of E2 was inserted into 2 different vectors and expressed as recombinant GST-E2 and recombinant MBP-E2 fusion protein, respectively. Two kinds of fusion proteins were tested with 30 CHIKV patient sera and 30 normal sera, respectively. Both proteins were detected by 25 patients sera (83.3%) and 1 normal serum (3.3%). This test showed a relatively high sensitivity and very high specificity of the recombinant E2 proteins to be used as diagnostic antigens against CHIKV infection. PMID:27180586

  5. Reexamination of alcohol dehydrogenase structural mutants in Drosophila using protein blotting

    SciTech Connect

    Hollocher, H.; Place, A.R.

    1987-06-01

    Using protein blotting and an immuno-overlay procedure, the authors have reexamined the cross-reacting material produced by ADH null-activity mutants generated with ethyl methanesulfonate (EMS). Of the 13 mutants, 11 have an immunodetectable polypeptide of wild-type size. The native and urea denatured isoelectric points (pI) establish that 7 of 13 of the mutations have no effect on protein charge. The electrophoretic mobilities of each variant on increasing percent acrylamide gels (Ferguson analysis), reveal that 9 of the 11 immunodetectable mutations have retained the ability form dimers under native conditions. None of the inactive mutant proteins has the ability to form the adduct-bound isozyme. The authors have found no correlation between protein pI and i vivo stability. The observed frequencies of specific charge class alterations do not dispute the propensity of G:A transitions previously found for EMS mutagenesis.

  6. 2-D Western blotting for evaluation of antibodies developed for detection of host cell protein.

    PubMed

    Berkelman, Tom; Harbers, Adriana; Bandhakavi, Sricharan

    2015-01-01

    Recombinant proteins generated for therapeutic use must be substantially free of residual host cell protein (HCP). The presence of host cell protein (HCP) is usually assayed by ELISA using a polyclonal antibody mixture raised against a population of proteins derived from the host cell background. This antibody should recognize as high a proportion as possible of the potential HCPs in a given sample. A recommended method for evaluating the assay involves two-dimensional electrophoretic separation followed by Western blotting.We present here a method using commercial anti-HCP antibody and samples derived from Chinese Hamster Ovary (CHO) cells. The 2-D electrophoresis procedure gives highly reproducible spot patterns and entire procedure can be completed in less than 2 days. Software analysis enables the straightforward generation of percent coverage values for the antibody when used to probe HCP-containing samples. PMID:25820736

  7. Food additives

    MedlinePlus

    Food additives are substances that become part of a food product when they are added during the processing or making of that food. "Direct" food additives are often added during processing to: Add nutrients ...

  8. Identification of Xanthomonas fragariae, Xanthomonas axonopodis pv. phaseoli, and Xanthomonas fuscans subsp. fuscans with novel markers and using a dot blot platform coupled with automatic data analysis.

    PubMed

    Albuquerque, Pedro; Caridade, Cristina M R; Marcal, Andre R S; Cruz, Joana; Cruz, Leonor; Santos, Catarina L; Mendes, Marta V; Tavares, Fernando

    2011-08-15

    Phytosanitary regulations and the provision of plant health certificates still rely mainly on long and laborious culture-based methods of diagnosis, which are frequently inconclusive. DNA-based methods of detection can circumvent many of the limitations of currently used screening methods, allowing a fast and accurate monitoring of samples. The genus Xanthomonas includes 13 phytopathogenic quarantine organisms for which improved methods of diagnosis are needed. In this work, we propose 21 new Xanthomonas-specific molecular markers, within loci coding for Xanthomonas-specific protein domains, useful for DNA-based methods of identification of xanthomonads. The specificity of these markers was assessed by a dot blot hybridization array using 23 non-Xanthomonas species, mostly soil dwelling and/or phytopathogens for the same host plants. In addition, the validation of these markers on 15 Xanthomonas spp. suggested species-specific hybridization patterns, which allowed discrimination among the different Xanthomonas species. Having in mind that DNA-based methods of diagnosis are particularly hampered for unsequenced species, namely, Xanthomonas fragariae, Xanthomonas axonopodis pv. phaseoli, and Xanthomonas fuscans subsp. fuscans, for which comparative genomics tools to search for DNA signatures are not yet applicable, emphasis was given to the selection of informative markers able to identify X. fragariae, X. axonopodis pv. phaseoli, and X. fuscans subsp. fuscans strains. In order to avoid inconsistencies due to operator-dependent interpretation of dot blot data, an image-processing algorithm was developed to analyze automatically the dot blot patterns. Ultimately, the proposed markers and the dot blot platform, coupled with automatic data analyses, have the potential to foster a thorough monitoring of phytopathogenic xanthomonads. PMID:21705524

  9. Mycoplasma agassizii strain variation and distinct host antibody responses explain differences between enzyme-linked immunosorbent assays and Western blot assays.

    PubMed

    Wendland, Lori D; Klein, Paul A; Jacobson, Elliott R; Brown, Mary B

    2010-11-01

    The precarious status of desert (Gopherus agassizii) and gopher (G. polyphemus) tortoises has resulted in conservation efforts that now include health assessment as an important component of management decision-making. Mycoplasmal upper respiratory tract disease (URTD) is one of very few diseases in chelonians for which comprehensive and rigorously validated diagnostic tests exist. In this study, serum samples obtained from eight Gopherus tortoises documented at necropsy to (i) be enzyme-linked immunosorbent assay (ELISA) seropositive using the PS6 antigen, (ii) be infected with Mycoplasma agassizii as indicated by direct isolation of the pathogen from the respiratory surfaces, and (iii) have histological lesions of mycoplasmal URTD were used to evaluate four distinct clinical isolates of M. agassizii as antigens for ELISA and Western blot analyses. Each animal sample reacted in the Western blot with its homologous M. agassizii strain, but recognition of heterologous M. agassizii strains was variable. Further, individual animals varied significantly with respect to the specific proteins recognized by the humoral immune response. An additional 114 Gopherus serum samples were evaluated using ELISA antigens prepared from the four distinct M. agassizii strains; A₄₀₅ values were significantly correlated (r² goodness of fit range, 0.708 to 0.771; P < 0.0001) for all antigens tested. The results confirm that strain variation is responsible for the observed differences between Western blot binding patterns. Thus, reliance on a single M. agassizii strain as an antigen in Western blot assays may provide false-negative results. This could have adverse consequences for the well-being of these environmentally sensitive hosts if false-negative animals were relocated to sites consisting of true-negative populations. PMID:20810678

  10. Mycoplasma agassizii Strain Variation and Distinct Host Antibody Responses Explain Differences between Enzyme-Linked Immunosorbent Assays and Western Blot Assays ▿

    PubMed Central

    Wendland, Lori D.; Klein, Paul A.; Jacobson, Elliott R.; Brown, Mary B.

    2010-01-01

    The precarious status of desert (Gopherus agassizii) and gopher (G. polyphemus) tortoises has resulted in conservation efforts that now include health assessment as an important component of management decision-making. Mycoplasmal upper respiratory tract disease (URTD) is one of very few diseases in chelonians for which comprehensive and rigorously validated diagnostic tests exist. In this study, serum samples obtained from eight Gopherus tortoises documented at necropsy to (i) be enzyme-linked immunosorbent assay (ELISA) seropositive using the PS6 antigen, (ii) be infected with Mycoplasma agassizii as indicated by direct isolation of the pathogen from the respiratory surfaces, and (iii) have histological lesions of mycoplasmal URTD were used to evaluate four distinct clinical isolates of M. agassizii as antigens for ELISA and Western blot analyses. Each animal sample reacted in the Western blot with its homologous M. agassizii strain, but recognition of heterologous M. agassizii strains was variable. Further, individual animals varied significantly with respect to the specific proteins recognized by the humoral immune response. An additional 114 Gopherus serum samples were evaluated using ELISA antigens prepared from the four distinct M. agassizii strains; A405 values were significantly correlated (r2 goodness of fit range, 0.708 to 0.771; P < 0.0001) for all antigens tested. The results confirm that strain variation is responsible for the observed differences between Western blot binding patterns. Thus, reliance on a single M. agassizii strain as an antigen in Western blot assays may provide false-negative results. This could have adverse consequences for the well-being of these environmentally sensitive hosts if false-negative animals were relocated to sites consisting of true-negative populations. PMID:20810678

  11. Comparative evaluation of western blotting in hepatic and pulmonary cystic echinococcosis.

    PubMed

    Akisu, C; Delibas, S B; Bicmen, C; Ozkoc, S; Aksoy, U; Turgay, N

    2006-12-01

    Many serological tests are widely used in the diagnosis of cystic echinococcosis (CE), caused by the larval stages of Echinococcus granulosus. The present study was carried for differentiation between hepatic and pulmonary cystic echinococcosis by Western Blotting (WB). A total of 121 sera from patients with hepatic CE (37), pulmonary CE (31) and controls (53; consisting of six healthy, seven Hymenolepis nana infection, 20 hepatic and 20 pulmonary diseases other than CE) were examined. In all of the CE patients, E. gronulosus infection was confirmed by surgical intervention. Sera were previously tested using IHA and ELISA to detect the E. gronulosus specific antibodies. Sera from hepatic cases of CE reacted with 16 polypeptides of 6-116 kDa and sera from pulmonary cases of CE reacted with 14 polypeptides of 4-130 kDa by Western Blotting. The WB test enabled the detection of antibodies in the hepatic CE samples for proteins of 24, 32 34, 44-46 and 52-54 kDa in molecular weight in 78.4%, 75.7%, 78.4% and 89.2% of the patients, respectively. In the pulmonary CE samples sera WB test enabled the detection of antibodies 24, 44-46, 100, 110, 116 and 120 124 kDa in molecular weight in 81.3%, 75.0%, 87.5%, 71.9%, 84.4% and 65.6% of the patients, respectively. We indicated that the antigenic components of high molecular weight can be good candidates for differentiation of hepatic CE from pulmonary CE. PMID:17285854

  12. Food additives

    PubMed Central

    Spencer, Michael

    1974-01-01

    Food additives are discussed from the food technology point of view. The reasons for their use are summarized: (1) to protect food from chemical and microbiological attack; (2) to even out seasonal supplies; (3) to improve their eating quality; (4) to improve their nutritional value. The various types of food additives are considered, e.g. colours, flavours, emulsifiers, bread and flour additives, preservatives, and nutritional additives. The paper concludes with consideration of those circumstances in which the use of additives is (a) justified and (b) unjustified. PMID:4467857

  13. Western blot analysis to illustrate relative control levels of the lac and ara promoters in Escherichia coli.

    PubMed

    Nielsen, Brent L; Willis, Van C; Lin, Chin-Yo

    2007-03-01

    The lactose operon and its control is a fundamental transcriptional regulatory concept presented in introductory and many advanced molecular biology courses. Much is known about the positive and negative control mechanisms that govern levels of expression of this operon. One basic principle that is taught about the lac operon is that it is "leaky," meaning that the transcriptional control of the operon is not 100% efficient and that in wild-type cells, transcription from the promoter is never completely "off," but there is always some basal transcription. In contrast, the arabinose operon is often used as an example of a tightly controlled operon, and transcription from the ara promoter is very low in the absence of inducer. The relative levels of control of these two operons can be illustrated using Western blots of proteins expressed in the presence and absence of the appropriate inducers and antibodies against the gene products. Different times of growth and the addition of inducer can also be examined. The results are very dramatic and help to reinforce the principles of promoter control. PMID:21591073

  14. The diagnosis of proventricular dilatation disease: use of a Western blot assay to detect antibodies against avian Borna virus.

    PubMed

    Villanueva, Itamar; Gray, Patricia; Mirhosseini, Negin; Payne, Susan; Hoppes, Sharman; Honkavuori, Kirsi S; Briese, Thomas; Turner, Debra; Tizard, Ian

    2010-07-14

    Avian Borna virus (ABV) has recently been shown to be the causal agent of proventricular dilatation disease (PDD) a lethal neurologic disease of captive psittacines and other birds. An immunoblot assay was used to detect the presence of antibodies against avian Borna virus in the serum of affected birds. A lysate from ABV-infected duck embryo fibroblasts served as a source of antigen. The assay was used to test for the presence of antibodies to ABV in 117 birds. Thirty of these birds had biopsy or necropsy-confirmed proventricular dilatation disease (PDD), while the remaining 87 birds were apparently healthy or were suffering from diseases other than PDD. Sera from 27 of the 30 PDD cases (90%) contained antibodies to ABV. Seventy-three (84%) of the apparently "healthy" birds were seronegative. Additionally, sera from seven macaws and one parrot trapped in the Peruvian Amazon were seronegative. Positive sera recognized the bornaviral nucleoprotein (N-protein). While the presence of antibodies to ABV largely corresponded with the development of clinical PDD, 14 apparently healthy normal birds possessed detectable antibodies to ABV. The existence of a carrier state was confirmed when 13 of 15 apparently healthy cockatiels were shown by PCR to have detectable ABV RNA in their feces. Western blot assays may be of significant assistance in diagnosing proventricular dilatation disease. Many apparently healthy birds may however be seronegative while, at the same time, shedding ABV in their feces. PMID:20036080

  15. Exposure to Sarcocystis spp. in horses from Spain determined by Western blot analysis using Sarcocystis neurona merozoites as heterologous antigen.

    PubMed

    Arias, M; Yeargan, M; Francisco, I; Dangoudoubiyam, S; Becerra, P; Francisco, R; Sánchez-Andrade, R; Paz-Silva, A; Howe, D K

    2012-04-30

    Horses serve as an intermediate host for several species of Sarcocystis, all of which utilize canids as the definitive host. Sarcocystis spp. infection and formation of latent sarcocysts in horses often appears to be subclinical, but morbidity can occur, especially when the parasite burden is large. A serological survey was conducted to determine the presence of antibodies against Sarcocystis spp. in seemingly healthy horses from the Galicia region of Spain. Western blot analyses using Sarcocystis neurona merozoites as heterologous antigen suggested greater than 80% seroprevalance of Sarcocystis spp. in a sample set of 138 horses. The serum samples were further tested with enzyme-linked immunosorbent assays (ELISAs) based on recombinant S. neurona-specific surface antigens (rSnSAGs). As expected for horses from the Eastern Hemisphere, less than 4% of the serum samples were positive when analyzed with either the rSnSAG2 or the rSnSAG4/3 ELISAs. An additional 246 horses were tested using the rSnSAG2 ELISA, which revealed that less than 3% of the 384 samples were seropositive. Collectively, the results of this serologic study suggested that a large proportion of horses from this region of Spain are exposed to Sarcocystis spp. Furthermore, the anti-Sarcocystis seroreactivity in these European horses could be clearly distinguished from anti-S. neurona antibodies using the rSnSAG2 and rSnSAG4/3 ELISAs. PMID:22019182

  16. Far-western blotting as a solution to the non-specificity of the anti-erythropoietin receptor antibody

    PubMed Central

    Fecková, Barbora; Kimáková, Patrícia; Ilkovičová, Lenka; Szentpéteriová, Erika; Debeljak, Nataša; Solárová, Zuzana; Sačková, Veronika; Šemeláková, Martina; Bhide, Mangesh; Solár, Peter

    2016-01-01

    The erythropoietin receptor (EpoR) is a member of the cytokine receptor family. The interaction between erythropoietin (Epo) and EpoR is important for the production and maturation of erythroid cells, resulting in the stimulation of hematopoiesis. The fact that EpoR was also detected in neoplastic cells has opened the question about the relevance of anemia treatment with recombinant Epo in cancer patients. Numerous studies have reported pro-stimulating and anti-apoptotic effects of Epo in cancer cells, thus demonstrating EpoR functionality in these cells. By contrast, a previous study claims the absence of EpoR in tumor cells. This apparent discrepancy is based, according to certain authors, on the use of non-specific anti-EpoR antibodies. With the aim of bypassing the direct detection of EpoR with an anti-EpoR antibody, the present authors propose a far-western blot methodology, which in addition, confirms the interaction of Epo with EpoR. Applying this technique, the presence of EpoR and its interaction with Epo in human ovarian adenocarcinoma A2780 and normal human umbilical vein endothelial cells was confirmed. Furthermore, modified immunoprecipitation of EpoR followed by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry analysis confirmed a 57 kDa protein as a human Epo-interacting protein in both cell lines. PMID:27446474

  17. Detection and typing of human papillomavirus using the Vira Type "in situ" kit: comparison with a conventional dot blot technique.

    PubMed Central

    Faulkner-Jones, B E; Bellomarino, V M; Borg, A J; Orzeszko, K; Garland, S M

    1990-01-01

    A new commercial kit (Vira Type "in situ", Life Technologies, Inc., Molecular Diagnostics Division, Guithersburg, Maryland, USA) for the detection of human papillomavirus (HPV) types 6, 11, 16, 18, 31, 33 and 35 in routinely processed human anogenital tissue was compared with a conventional dot blot assay for HPV 6, 11, 16 and 18. Both systems use double-stranded genomic DNA probes for the detection of type specific HPV DNA. The probes used on the dot blots were labelled with 32P and visualised autoradiographically. The Vira Type probes were labelled with biotin and visualised using a streptavidin-alkaline phosphatase conjugate with NBT-BCIP substrate. Biopsy specimens from the cervix, vagina, and vulva of 46 women were processed by both methods and compared. The histological diagnoses ranged from benign changes, to dysplasia, and invasive carcinoma. Overall, 50% of biopsy specimens were positive for HPV DNA by dot blot hybridisation; only 39% were positive by Vira Type in situ hybridisation. Three of the specimens positive by the Vira Type "in situ" kit showed no cross hybridisation and were the same HPV type as the dot blot. A further 13 showed hybridisation, but the showed cross hybridisation, but the to the dot blot results. One biopsy specimen was positive for different HPV types by the two tests and one was positive by Vira Type and negative by dot blot. Six biopsy specimens were negative by Vira Type but positive by dot blot. It is concluded that the Vira Type "in situ" kit has a similar specificity but lower sensitivity than the dot blot hybridisation method for the detection of HPV DNA. Images PMID:2175755

  18. V3 Stain-free Workflow for a Practical, Convenient, and Reliable Total Protein Loading Control in Western Blotting

    PubMed Central

    Posch, Anton; Kohn, Jonathan; Oh, Kenneth; Hammond, Matt; Liu, Ning

    2013-01-01

    The western blot is a very useful and widely adopted lab technique, but its execution is challenging. The workflow is often characterized as a "black box" because an experimentalist does not know if it has been performed successfully until the last of several steps. Moreover, the quality of western blot data is sometimes challenged due to a lack of effective quality control tools in place throughout the western blotting process. Here we describe the V3 western workflow, which applies stain-free technology to address the major concerns associated with the traditional western blot protocol. This workflow allows researchers: 1) to run a gel in about 20-30 min; 2) to visualize sample separation quality within 5 min after the gel run; 3) to transfer proteins in 3-10 min; 4) to verify transfer efficiency quantitatively; and most importantly 5) to validate changes in the level of the protein of interest using total protein loading control. This novel approach eliminates the need of stripping and reprobing the blot for housekeeping proteins such as β-actin, β-tubulin, GAPDH, etc. The V3 stain-free workflow makes the western blot process faster, transparent, more quantitative and reliable. PMID:24429481

  19. Performance of PCR-reverse blot hybridization assay for detection of rifampicin-resistant Mycobacterium leprae.

    PubMed

    Wang, Hye-young; Kim, Hyunjung; Kim, Yeun; Bang, Hyeeun; Kim, Jong-Pill; Hwang, Joo Hwan; Cho, Sang-Nae; Kim, Tae Ue; Lee, Hyeyoung

    2015-10-01

    Drug resistance in Mycobacterium leprae is a significant problem in countries where leprosy is endemic. A sensitive, specific, and high-throughput reverse blot hybridization assay (REBA) for the detection of genotypic resistance to rifampicin (RIF) was designed and evaluated. It has been shown that resistance to RIF in M. leprae involves mutations in the rpoB gene encoding the -subunit of the RNA polymerase. The PCR-REBA simultaneously detects both 6 wild-type regions and 5 different mutations (507 AGC, 513 GTG, 516 TAT, 531 ATG, and 531 TTC) including the most prevalent mutations at positions 507 and 531. Thirty-one clinical isolates provided by Korea Institute of Hansen-s Disease were analyzed by PCR-REBA with RIF resistance of rpoB gene. As a result, missense mutations at codons 507 AGC and 531 ATG with 2-nucleotide substitutions were found in one sample, and a missense mutation at codon 516 TAT and ΔWT6 (deletion of 530-534) was found in another sample. These cases were confirmed by DNA sequence analysis. This rapid, simple, and highly sensitive assay provides a practical alternative to sequencing for genotypic evaluation of RIF resistance in M. leprae. PMID:26428919

  20. Pairwise detection of site-specific receptor phosphorylations using single-molecule blotting

    PubMed Central

    Kim, Kyung Lock; Kim, Daehyung; Lee, Seongsil; Kim, Su-Jeong; Noh, Jung Eun; Kim, Joung-Hun; Chae, Young Chan; Lee, Jong-Bong; Ryu, Sung Ho

    2016-01-01

    Post-translational modifications (PTMs) of receptor tyrosine kinases (RTKs) at the plasma membrane (PM) determine the signal transduction efficacy alone and in combination. However, current approaches to identify PTMs provide ensemble results, inherently overlooking combinatorial PTMs in a single polypeptide molecule. Here, we describe a single-molecule blotting (SiMBlot) assay that combines biotinylation of cell surface receptors with single-molecule fluorescence microscopy. This method enables quantitative measurement of the phosphorylation status of individual membrane receptor molecules and colocalization analysis of multiple immunofluorescence signals to directly visualize pairwise site-specific phosphorylation patterns at the single-molecule level. Strikingly, application of SiMBlot to study ligand-dependent epidermal growth factor receptor (EGFR) phosphorylation, which is widely thought to be multi-phosphorylated, reveals that EGFR on cell membranes is hardly multi-phosphorylated, unlike in vitro autophosphorylated EGFR. Therefore, we expect SiMBlot to aid understanding of vast combinatorial PTM patterns, which are concealed in ensemble methods, and to broaden knowledge of RTK signaling. PMID:27009355

  1. Digital chemiluminescence imaging of DNA sequencing blots using a charge-coupled device camera.

    PubMed Central

    Karger, A E; Weiss, R; Gesteland, R F

    1992-01-01

    Digital chemiluminescence imaging with a cryogenically cooled charge-coupled device (CCD) camera is used to visualize DNA sequencing fragments covalently bound to a blotting membrane. The detection is based on DNA hybridization with an alkaline phosphatase(AP) labeled oligodeoxyribonucleotide probe and AP triggered chemiluminescence of the substrate 3-(2'-spiro-adamantane)-4-methoxy-4-(3"-phosphoryloxy)phenyl- 1,2-dioxetane (AMPPD). The detection using a direct AP-oligonucleotide conjugate is compared to the secondary detection of biotinylated oligonucleotides with respect to their sensitivity and nonspecific binding to the nylon membrane by quantitative imaging. Using the direct oligonucleotide-AP conjugate as a hybridization probe, sub-attomol (0.5 pg of 2.7 kb pUC plasmid DNA) quantities of membrane bound DNA are detectable with 30 min CCD exposures. Detection using the biotinylated probe in combination with streptavidin-AP was found to be background limited by nonspecific binding of streptavidin-AP and the oligo(biotin-11-dUTP) label in equal proportions. In contrast, the nonspecific background of AP-labeled oligonucleotide is indistinguishable from that seen with 5'-32P-label, in that respect making AP an ideal enzymatic label. The effect of hybridization time, probe concentration, and presence of luminescence enhancers on the detection of plasmid DNA were investigated. Images PMID:1480487

  2. Retrospective Study of Hemoparasites in Cattle in Southern Italy by Reverse Line Blot Hybridization

    PubMed Central

    CECI, Luigi; IARUSSI, Fabrizio; GRECO, Beatrice; LACINIO, Rosanna; FORNELLI, Stefania; CARELLI, Grazia

    2014-01-01

    ABSTRACT Tick-borne diseases are widespread in tropical and temperate regions and are responsible for important economic losses in those areas. In order to assess the presence and prevalence of various pathogens in southern Italy, we retrospectively analyzed cattle blood samples collected for a previous study in 2000 using reverse line blot (RLB) hybridization. The study had been carried out in three regions of southern Italy on 1,500 randomly selected and apparently healthy adult cattle. RLB showed that 43.7% of the cattle were positive for nine different species of hemoparasites with either a single infection or a mixed infection. Theileria buffeli was the most common species found, being present in 27.3% of the animals, followed by Anaplasma marginale in 18.1%, Anaplasma centrale in 13.8%, Babesia bigemina and Anaplasma bovis in 4.2%, Anaplasma phagocytophilum in 1.7%, Babesia bovis in 1.6%, Babesia major in 0.2% and Babesia divergens in 0.1%. Complete blood counts showed different degrees of anemia in 363 animals (24.2%) and of these, 169 were RLB-positive for at least one pathogen. Among the ticks that were collected from the cattle, the following species were identified: Rhipicephalus bursa, Ixodes ricinus, Hyalomma marginatum, Boophilus annulatus, Dermacentor marginatus and Haemaphysalis (sulcata, parva, inermis and punctata). The results obtained confirmed the spread of endemic tick-borne pathogens in the regions studied. PMID:24614604

  3. 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.

  4. Detection of strawberry vein banding virus by polymerase chain reaction and dot blot hybridization.

    PubMed

    Mráz, I; Petrzik, K; Fránová-Honetslegrová, J; Síp, M

    1997-08-01

    Strawberry vein banding virus (SVBV) is one of seventeen members of the family Caulimoviridae. Natural infection with the virus is known in Fragaria species only. Infections caused by SVBV are often symptomless (1), but their significance increases in mixed infections with strawberry crinkle or strawberry latent C viruses (2,3). This virus has been originally found on strawberries in USA and firstly described by Frazier (4), but it is probably world-wide distributed by planting or breeding materials. SVBV has been observed on cultivated strawberries in North America, Australia, Brazil, Japan (5) and recently in Europe (6,7). The concentration of SVBV in infected plants is usually very low. Its detection by ELISA is impossible because of lack of specific antibodies. Evidence of the caulimovirus nature of SVBV has been confirmed by its circular dsDNA genome, shape and size of viral particles (8), presence of cytoplasmic inclusion bodies typical for caulimoviruses, and distant serological relationship with cauliflower mosaic virus (CaMV, 9). In this paper we present detection of SVBV by combination of two detection methods--polymerase chain reaction (PCR) and dot blot hybridization with a non-radioactive probe. PMID:9391655

  5. The blot rolling assay: a method for identifying adhesion molecules mediating binding under shear conditions.

    PubMed

    Sackstein, Robert; Fuhlbrigge, Robert

    2006-01-01

    Adhesive interactions of cells with blood vessel walls under flow conditions are critical to a variety of processes, including hemostasis, leukocyte trafficking, tumor metastasis, and atherosclerosis. We have developed a new technique for the observation of binding interactions under shear, which we have termed the "blot rolling assay." In this method, molecules in a complex mixture are resolved by gel electrophoresis and transferred to a membrane. This membrane can be rendered semitransparent and incorporated into a parallel-plate flow chamber apparatus. Cells or particles bearing adhesion proteins of interest are then introduced into the chamber under controlled flow, and their interactions with individual components of the immobilized substrates can be visualized in real time. The substrate molecules can be identified by staining with specific antibodies or by excising the relevant band(s) and performing mass spectrometry or microsequencing of the isolated material. Thus, this method allows for the identification, within a complex mixture and without previous isolation or purification, of both known and novel adhesion molecules capable of binding under shear conditions. PMID:16799202

  6. [Evaluation of IHA, ELISA and Western Blot tests in diagnosis of pulmonary cystic hidatidosis].

    PubMed

    Akisu, Ciler; Bayram Delibaş, Songül; Yuncu, Gökhan; Aksoy, Umit; Ozkoç, Soykan; Biçmen, Can; Sevinç, Serpil; Yaldiz, Sadik

    2005-01-01

    Pulmonary cystic hidatidosis caused by the larval stages of Echinococcus granulosus is a common parasitic disease in Turkey and throughout the world. In this study IHA, ELISA and Western Blot (WB) tests were performed with a panel of 59 sera from 31 surgically confirmed pulmonary cystic hidatidosis patients, 18 patients with pulmonary disease other than cystic hidatidosis and 10 healthy individual. The overall sensitivity of the IHA, ELISA and WB tests used for the serodiagnosis of pulmonary cystic hidatidosis were found as 96.7%, 87.1%, 100% and the specificities were 82.2%, 89.2% and %85.7, respectively. Using the WB test 8-12 kDa, 24 kDa and 124 kDa bands were detected as valuable for surgically confirmed patients' sera. One or more of these bands were also detected in sera of four patients with other pulmonary diseases false-positively. In conclusion conventional serologic test like IHA and ELISA is valuable in diagnosis of pulmonary cystic hidatidosis, also evaluation of some specific bands in WB would contribute to the diagnosis. PMID:16100652

  7. Food additives.

    PubMed

    Berglund, F

    1978-01-01

    The use of additives to food fulfils many purposes, as shown by the index issued by the Codex Committee on Food Additives: Acids, bases and salts; Preservatives, Antioxidants and antioxidant synergists; Anticaking agents; Colours; Emulfifiers; Thickening agents; Flour-treatment agents; Extraction solvents; Carrier solvents; Flavours (synthetic); Flavour enhancers; Non-nutritive sweeteners; Processing aids; Enzyme preparations. Many additives occur naturally in foods, but this does not exclude toxicity at higher levels. Some food additives are nutrients, or even essential nutritents, e.g. NaCl. Examples are known of food additives causing toxicity in man even when used according to regulations, e.g. cobalt in beer. In other instances, poisoning has been due to carry-over, e.g. by nitrate in cheese whey - when used for artificial feed for infants. Poisonings also occur as the result of the permitted substance being added at too high levels, by accident or carelessness, e.g. nitrite in fish. Finally, there are examples of hypersensitivity to food additives, e.g. to tartrazine and other food colours. The toxicological evaluation, based on animal feeding studies, may be complicated by impurities, e.g. orthotoluene-sulfonamide in saccharin; by transformation or disappearance of the additive in food processing in storage, e.g. bisulfite in raisins; by reaction products with food constituents, e.g. formation of ethylurethane from diethyl pyrocarbonate; by metabolic transformation products, e.g. formation in the gut of cyclohexylamine from cyclamate. Metabolic end products may differ in experimental animals and in man: guanylic acid and inosinic acid are metabolized to allantoin in the rat but to uric acid in man. The magnitude of the safety margin in man of the Acceptable Daily Intake (ADI) is not identical to the "safety factor" used when calculating the ADI. The symptoms of Chinese Restaurant Syndrome, although not hazardous, furthermore illustrate that the whole ADI

  8. Evaluation of ELISA coupled with Western blot as a surveillance tool for Trichinella infection in wild boar (Sus scrofa).

    PubMed

    Cuttell, Leigh; Gómez-Morales, Maria Angeles; Cookson, Beth; Adams, Peter J; Reid, Simon A; Vanderlinde, Paul B; Jackson, Louise A; Gray, C; Traub, Rebecca J

    2014-01-31

    Trichinella surveillance in wildlife relies on muscle digestion of large samples which are logistically difficult to store and transport in remote and tropical regions as well as labour-intensive to process. Serological methods such as enzyme-linked immunosorbent assays (ELISAs) offer rapid, cost-effective alternatives for surveillance but should be paired with additional tests because of the high false-positive rates encountered in wildlife. We investigated the utility of ELISAs coupled with Western blot (WB) in providing evidence of Trichinella exposure or infection in wild boar. Serum samples were collected from 673 wild boar from a high- and low-risk region for Trichinella introduction within mainland Australia, which is considered Trichinella-free. Sera were examined using both an 'in-house' and a commercially available indirect-ELISA that used excretory-secretory (E/S) antigens. Cut-off values for positive results were determined using sera from the low-risk population. All wild boar from the high-risk region (352) and 139/321 (43.3%) of the wild boar from the low-risk region were tested by artificial digestion. Testing by Western blot using E/S antigens, and a Trichinella-specific real-time PCR was also carried out on all ELISA-positive samples. The two ELISAs correctly classified all positive controls as well as one naturally infected wild boar from Gabba Island in the Torres Strait. In both the high- and low-risk populations, the ELISA results showed substantial agreement (k-value=0.66) that increased to very good (k-value=0.82) when WB-positive only samples were compared. The results of testing sera collected from the Australian mainland showed the Trichinella seroprevalence was 3.5% (95% C.I. 0.0-8.0) and 2.3% (95% C.I. 0.0-5.6) using the in-house and commercial ELISA coupled with WB respectively. These estimates were significantly higher (P<0.05) than the artificial digestion estimate of 0.0% (95% C.I. 0.0-1.1). Real-time PCR testing of muscle from

  9. Application of reverse dot blot hybridization to simultaneous detection and identification of harmful algae.

    PubMed

    Chen, Guo Fu; Zhang, Chun Yun; Wang, Yuan Yuan; Chen, Wen

    2015-07-01

    Warning and monitoring projects of harmful algal blooms require simple and rapid methods for simultaneous and accurate detection and identification of causative algae present in the environmental samples. Here, reverse dot blot hybridization (RDBH) was employed to simultaneously detect several harmful algae by using five representative bloom-forming microalgae along the Chinese coast. A set of specific probes for RDBH were developed by PCR, cloning, and sequencing of the internal transcribed spacer (ITS), alignment analysis, and probe design. Each probe was oligo (dT)-tailed and spotted onto positively charged nylon membrane to make up a low-density oligonucleotide array. Universal primers designed within the conserved regions were used to amplify the ITS sequences by using genomic DNA of target as templates. The digoxigenin (Dig)-labeled PCR products were denatured and then hybridized to the oligonucleotide array. The array produced a unique hybridization pattern for each target species differentiating them from each other. The preparations of oligonucleotide array and hybridization conditions were optimized. The developed RDBH demonstrated a detection limit up to 10 cells. The detection performance of RDBH was relatively stable and not affected by non-target species and the fixation time of target species over at least 30 days. The RDBH could recover all the target species from the simulated field samples and target species confirmed by the subsequent microscopy examination in the environmental samples. These results indicate that RDBH can be a new technical platform for parallel discrimination of harmful algae and is promising for environmental monitoring of these microorganisms. PMID:25731086

  10. Multiplex PCR and Reverse Line Blot Hybridization Assay (mPCR/RLB)

    PubMed Central

    O'Sullivan, Matthew V. N.; Zhou, Fei; Sintchenko, Vitali; Kong, Fanrong; Gilbert, Gwendolyn L.

    2011-01-01

    Multiplex PCR/Reverse Line Blot Hybridization assay allows the detection of up to 43 molecular targets in 43 samples using one multiplex PCR reaction followed by probe hybridization on a nylon membrane, which is re-usable. Probes are 5' amine modified to allow fixation to the membrane. Primers are 5' biotin modified which allows detection of hybridized PCR products using streptavidin-peroxidase and a chemiluminescent substrate via photosensitive film. With low setup and consumable costs, this technique is inexpensive (approximately US$2 per sample), high throughput (multiple membranes can be processed simultaneously) and has a short turnaround time (approximately 10 hours). The technique can be utilized in a number of ways. Multiple probes can be designed to detect sequence variation within a single amplified product, or multiple products can be amplified simultaneously, with one (or more) probes used for subsequent detection. A combination of both approaches can also be used within a single assay. The ability to include multiple probes for a single target sequence makes the assay highly specific. Published applications of mPCR/RLB include detection of antibiotic resistance genes1,2, typing of methicillin-resistant Staphylococcus aureus3-5 and Salmonella sp6, molecular serotyping of Streptococcus pneumoniae7,8, Streptococcus agalactiae9 and enteroviruses10,11, identification of Mycobacterium sp12, detection of genital13-15 and respiratory tract16 and other17 pathogens and detection and identification of mollicutes18. However, the versatility of the technique means the applications are virtually limitless and not restricted to molecular analysis of micro-organisms. The five steps in mPCR/RLB are a) Primer and Probe design, b) DNA extraction and PCR amplification c) Preparation of the membrane, d) Hybridization and detection, and e) Regeneration of the Membrane. PMID:21847083

  11. Evaluation of a Western Blot Test in an Outbreak of Acute Pulmonary Histoplasmosis

    PubMed Central

    Pizzini, Claudia V.; Zancopé-Oliveira, Rosely M.; Reiss, Errol; Hajjeh, Rana; Kaufman, Leo; Peralta, José Mauro

    1999-01-01

    A western blot (WB) test was evaluated for detection of antibodies against native glycosylated and chemically deglycosylated M and H antigens of Histoplasma capsulatum in serum obtained from patients during the acute phase of pulmonary histoplasmosis that occurred during an outbreak. Of 275 serum samples tested by immunodiffusion and complement fixation (CF) samples from 40 patients affected during this outbreak and from 37 negative controls were tested by WB test. A group of patients whose sera were negative for CF antibodies and precipitins early in the acute stage of histoplasmosis but who all seroconverted during convalescence 6 weeks later were tested with the WB test. Antibodies against untreated H and M antigens were detected at a 1:100 dilution by WB test in 45% of the 20 acute-phase serum samples and in all 20 of the convalescent-phase specimens. The WB test’s sensitivity for acute-phase specimens increased to 90% (18 of 20 specimens) when H and M antigens were treated by periodate oxidation to inactivate susceptible carbohydrate epitopes. When native glycosylated antigens were used in the WB test, positive reactions were observed in negative control serum specimens (3 of 37 specimens; 8%) and in serum specimens obtained from asymptomatic persons screened as part of the outbreak investigation (13 of 20 specimens; 65%). These positive reactions were also attributed to glycosidic epitopes since the specificity of the WB test increased from 78 to 100% when periodate-treated H and M antigens were used. WB test with deglycosylated H and M antigens of histoplasmin provides a rapid, sensitive, and specific test to diagnose acute pulmonary histoplasmosis before precipitins can be detected. PMID:9874658

  12. Enzyme-linked immunoelectrotransfer blot test for diagnosis of human hydatid disease.

    PubMed Central

    Verastegui, M; Moro, P; Guevara, A; Rodriguez, T; Miranda, E; Gilman, R H

    1992-01-01

    Sera from 71 patients with surgically confirmed hydatid disease (which is caused by Echinococcus granulosus) were studied by an enzyme-linked immunoelectrotransfer blot (EITB) assay. Sera from patients either with other cestode infections or with another illness were used as controls. Results of the EITB test for hydatidosis were compared with those of the double-diffusion (DD5) test and an enzyme-linked immunosorbent assay (ELISA). In the EITB assay with bovine lyophilized hydatid fluid, three antigen bands of 8, 16, and 21 kDa were diagnostically important. The sensitivity of the assay by using these antigen bands was 80% for hepatic cysts, 56% for pulmonary cysts, and 56% for cysts located in multiple organs. In sera from controls, the specificity of the EITB assay was 100%. Cross-reactions to the 8-, 16-, and 21-kDa bands occurred, respectively, in 12, 4, and 4% of sera from patients with cysticercosis. No cross-reactions were noted in patients infected with Hymenolepis nana. The ELISA in which swine hydatid fluid was used as the antigen was as sensitive as the EITB test but was less specific (80%) and frequently cross-reacted with sera from patients with other cestode infections. The sensitivity of the DD5 test, which uses sheep hydatid fluid, was low (47%) , but its specificity was as high as that of the EITB assay. However, in patients with cysticercosis, cross-reactions were observed in 23% of sera tested. Despite the higher sensitivity found with the EITB assay, 23% (n = 5) of the serum samples that were positive by the DD5 test were not detected by the EITB assay. The EITB assay offers greater sensitivity and specificity than do the ELISA and the DD5 test. The highest proportion of hydatid cases is detected when the EITB and DD5 tests are run simultaneously. Images PMID:1624574

  13. Validity of the Enzyme-linked Immunoelectrotransfer Blot (EITB) for naturally acquired porcine cysticercosis.

    PubMed

    Jayashi, César M; Gonzalez, Armando E; Castillo Neyra, Ricardo; Rodríguez, Silvia; García, Hector H; Lightowlers, Marshall W

    2014-01-17

    The Enzyme-linked Immunoelectrotransfer Blot (EITB) has been used widely as a screening test for Taenia solium cysticercosis in swine. However, the relation between seropositivity and infection in pig populations from endemic areas has not been well defined. The aim of this study is to relate EITB seropositivity with infection and infection burden, analyse the trade-off between sensitivity and specificity with various cut-off points for the EITB assay, and finally describe the serology changes in a cohort of rural pigs raised under natural conditions. A group of 107 pigs that were used as controls during a vaccination field trial in Peru was our study population. The prevalence of porcine cysticercosis determined by necropsy examination was 16.82% (18/107) in these animals. Using EITB reactivity to ≥ 1 band as a cut-off point for the assay, the sensitivity was 88.89% (65.29-98.62, 95% CI) and the specificity was 48.31% (37.59-59.16, 95% CI). Comparing other cut-off points, involving up to as many as 7 reactive bands, a reactivity of ≥ 3 bands provided the best trade-offs in sensitivity and specificity. Using this cut-off point for the assay, the sensitivity was 77.77% (52.36-93.59, 95% CI) and the specificity was 76.40% (66.22-84.76, 95% CI). A significant association was found between cyst counts over 100 cysts and reactivity to ≥ 3 bands in the EITB assay (Fisher's exact test, p<0.05). The results of this study suggest that the use of the EITB assay to study porcine cysticercosis may require setting different cut-offs under field and experimental conditions, and depending upon the objective of the screening process. PMID:24183647

  14. Development of a dot blot assay using gene probes for the detection of enteroviruses in water

    SciTech Connect

    Margolin, A.B.

    1986-01-01

    Enteric viruses are viruses which replicate in the intestinal tract of man and animals. One mode of transmission for enteric viruses is the fecal-oral route. Drinking water which has been contaminated with sewage or sewage effluent has been implicated as a means for the spread of enteric viruses. Current methods for the detection of enteric viruses in water requires the use of animal cell culture. This technique has several drawbacks. More rapid techniques, such as fluorescent antibody or radioimmunoassay do not have the needed sensitivity to detect the low levels of virus found in contaminated water. An alternative technique for the detection of viruses in water was sought. Recent advances in recombinant DNA technology now makes it possible to detect viruses without the use of cell culture or antibodies. Gene probes that hybridize to the RNA of poliovirus and hepatitis A virus were tested for their ability to detect different enteric viruses. The probes were labeled with /sup 32/P dCTP and /sup 32/P dATP to a specific activity greater then 1.0 x 10/sup 9/ cpm/ug DNA. One infectious unit of poliovirus and hepatitis A virus was detected using labeled cDNA probes. Upon comparison, the dot blot assay was as sensitive as tissue culture for the detection of poliovirus in beef extract, secondary effluent, and tap water. Environmental samples, such as secondary effluent, reclaimed wastewater and unchlorinated drinking water were also assayed for poliovirus and hepatitis A virus with the use of gene probes. The results presented here offer an alternative method for screening water samples for the presence of enteric viruses.

  15. Immunodot blot assay to detect Helicobacter pylori using monoclonal antibodies against the 26 kDa protein.

    PubMed

    Amini Najafabadi, Hossein; Paknejad, Maliheh; Farshad, Shohreh; Mohammadian, Taher; Seyyed Ebrahimi, Shadi Sadat; Amini Najafabadi, Azadeh

    2012-12-01

    Development of a specific immunoassay to detect Helicobacter pylori infection in stool samples requires monoclonal antibody against the specific antigen. The aims of this study were to establish monoclonal antibodies against the 26 kDa protein of H. pylori and develop an immunodot blot for their application to recognize H. pylori infection using stool samples. Mice were immunized intraperitoneally with homogenized gel containing the 26 kDa band of cell surface proteins of H. pylori in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The monoclonal antibodies were produced using the hybridoma technique. Reactivity of monoclonal antibodies was tested with the purified 26 kDa antigen and cell surface proteins from cultured H. pylori by ELISA. Furthermore reactivity of monoclonal antibodies was tested on negative and positive stool samples for H. pylori and suspensions of several major bacteria in stool by immunodot blot assay. Five stable hybridoma monoclones were obtained. The concordant reactivity of the monoclonal antibodies with H. pylori present in the stool samples, which had been tested previously using an ACON ELISA kit for H. pylori stool antigen testing, and unreactivity with several different major fecal bacteria in immunodot blotting indicates high specificity of the immunodot blot based on the reaction of produced monoclonal antibodies with the H. pylori antigen in stools. The findings indicate that the novel immunodot blot developed based on new monoclonal antibodies for stool antigens would be useful as a noninvasive method of diagnosing H. pylori infection. PMID:23244318

  16. Identification of α1-Antitrypsin as a Potential Candidate for Internal Control for Human Synovial Fluid in Western Blot

    PubMed Central

    Wang, Shaowei; Zhou, Jingming; Wei, Xiaochun; Li, Pengcui; Li, Kai; Wang, Dongming; Wei, Fangyuan; Zhang, Jianzhong; Wei, Lei

    2015-01-01

    Western blot of synovial fluid has been widely used for osteoarthritis (OA) research and diagnosis, but there is no ideal loading control for this purpose. Although β-actin is extensively used as loading control in western blot, it is not suitable for synovial fluid because it is not required in synovial fluid as a cytoskeletal protein. A good loading control for synovial fluid in OA studies should have unchanged content in synovial fluids from normal and OA groups, because synovial fluid protein content can vary with changes in synovial vascular permeability with OA onset. In this study, we explore the potential of using α1-antitripsin (A1AT) as loading control for OA synovial fluid in western blot. A1AT level is elevated in inflammatory conditions such as rheumatoid arthritis (RA). Unlike RA, OA is a non-inflammation disease, which does not induce A1AT. In this study, we identified A1AT as an abundant component of synovial fluid by Mass Spectrometry and confirmed that the level of A1AT is relative constant between human OA and normal synovial fluid by western blot and ELISA. Hence, we proposed that A1AT may be a good loading control for western blot in human OA synovial fluid studies provided that pathological conditions such as RA or A1AT deficiency associated liver or lung diseases are excluded. PMID:26594594

  17. Qualitative and quantitative evaluation of Simon™, a new CE-based automated Western blot system as applied to vaccine development.

    PubMed

    Rustandi, Richard R; Loughney, John W; Hamm, Melissa; Hamm, Christopher; Lancaster, Catherine; Mach, Anna; Ha, Sha

    2012-09-01

    Many CE-based technologies such as imaged capillary IEF, CE-SDS, CZE, and MEKC are well established for analyzing proteins, viruses, or other biomolecules such as polysaccharides. For example, imaged capillary isoelectric focusing (charge-based protein separation) and CE-SDS (size-based protein separation) are standard replacement methods in biopharmaceutical industries for tedious and labor intensive IEF and SDS-PAGE methods, respectively. Another important analytical tool for protein characterization is a Western blot, where after size-based separation in SDS-PAGE the proteins are transferred to a membrane and blotted with specific monoclonal or polyclonal antibodies. Western blotting analysis is applied in many areas such as biomarker research, therapeutic target identification, and vaccine development. Currently, the procedure is very manual, laborious, and time consuming. Here, we evaluate a new technology called Simple Western™ (or Simon™) for performing automated Western analysis. This new technology is based on CE-SDS where the separated proteins are attached to the wall of capillary by a proprietary photo activated chemical crosslink. Subsequent blotting is done automatically by incubating and washing the capillary with primary and secondary antibodies conjugated with horseradish peroxidase and detected with chemiluminescence. Typically, Western blots are not quantitative, hence we also evaluated the quantitative aspect of this new technology. We demonstrate that Simon™ can quantitate specific components in one of our vaccine candidates and it provides good reproducibility and intermediate precision with CV <10%. PMID:22965727

  18. Potlining Additives

    SciTech Connect

    Rudolf Keller

    2004-08-10

    In this project, a concept to improve the performance of aluminum production cells by introducing potlining additives was examined and tested. Boron oxide was added to cathode blocks, and titanium was dissolved in the metal pool; this resulted in the formation of titanium diboride and caused the molten aluminum to wet the carbonaceous cathode surface. Such wetting reportedly leads to operational improvements and extended cell life. In addition, boron oxide suppresses cyanide formation. This final report presents and discusses the results of this project. Substantial economic benefits for the practical implementation of the technology are projected, especially for modern cells with graphitized blocks. For example, with an energy savings of about 5% and an increase in pot life from 1500 to 2500 days, a cost savings of $ 0.023 per pound of aluminum produced is projected for a 200 kA pot.

  19. Phosphazene additives

    SciTech Connect

    Harrup, Mason K; Rollins, Harry W

    2013-11-26

    An additive comprising a phosphazene compound that has at least two reactive functional groups and at least one capping functional group bonded to phosphorus atoms of the phosphazene compound. One of the at least two reactive functional groups is configured to react with cellulose and the other of the at least two reactive functional groups is configured to react with a resin, such as an amine resin of a polycarboxylic acid resin. The at least one capping functional group is selected from the group consisting of a short chain ether group, an alkoxy group, or an aryloxy group. Also disclosed are an additive-resin admixture, a method of treating a wood product, and a wood product.

  20. Genotyping of 27 human papillomavirus types by using L1 consensus PCR products by a single-hybridization, reverse line blot detection method.

    PubMed

    Gravitt, P E; Peyton, C L; Apple, R J; Wheeler, C M

    1998-10-01

    Amplification of human papillomavirus (HPV) DNA by L1 consensus primer systems (e.g., MY09/11 or GP5(+)/6(+)) can detect as few as 10 to 100 molecules of HPV targets from a genital sample. However, genotype determination by dot blot hybridization is laborious and requires at least 27 separate hybridizations for substantive HPV-type discrimination. A reverse blot method was developed which employs a biotin-labeled PCR product hybridized to an array of immobilized oligonucleotide probes. By the reverse blot strip analysis, genotype discrimination of multiple HPV types can be accomplished in a single hybridization and wash cycle. Twenty-seven HPV probe mixes, two control probe concentrations, and a single reference line were immobilized to 75- by 6-mm nylon strips. Each individual probe line contained a mixture of two bovine serum albumin-conjugated oligonucleotide probes specific to a unique HPV genotype. The genotype spectrum discriminated on this strip includes the high-risk, or cancer-associated, HPV genotypes 16, 18, 26, 31, 33, 35, 39, 45, 51, 52, 55, 56, 58, 59, 68 (ME180), MM4 (W13B), MM7 (P291), and MM9 (P238A) and the low-risk, or non-cancer-associated, genotypes 6, 11, 40, 42, 53, 54, 57, 66, and MM8 (P155). In addition, two concentrations of beta-globin probes allowed for assessment of individual specimen adequacy following amplification. We have evaluated the performance of the strip method relative to that of a previously reported dot blot format (H. M. Bauer et al., p. 132-152, in C. S. Herrington and J. O. D. McGee (ed.), Diagnostic Molecular Pathology: a Practical Approach, (1992), by testing 328 cervical swab samples collected in Digene specimen transport medium (Digene Diagnostics, Silver Spring, Md.). We show excellent agreement between the two detection formats, with 92% concordance for HPV positivity (kappa = 0.78, P < 0.001). Nearly all of the discrepant HPV-positive samples resulted from weak signals and can be attributed to sampling error from

  1. Enrichment of PrPSc in Formalin Fixed Paraffin Embedded Tissues Prior to Analysis by Western Blot

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Diagnosis of prion disease is primarily through immunodetection of the infectious agent. Typically, 2 distinct procedures are recommended for a definitive diagnosis with immunohistochemistry and Western blot providing the most information as to the specific isolate in question. In the past these app...

  2. Identification of species in tribe Brassiceae by dot-blot hybridization using species-specific ITS1 probes.

    PubMed

    Tonosaki, K; Nishio, Takeshi

    2010-10-01

    Simple, reliable methods for identification of species are required for management of many species and lines in a plant gene bank. Species-specific probes were designed from published sequences of the ITS1 region in rDNA of 16 species in Brassica and its related genera, and used as probes for dot-blot hybridization with plant genomic DNA. All the probes detected species-specific signals at dot-blots of genomic DNAs of the 16 species in Brassica, Diplotaxis, Eruca, and Raphanus. Signals of the Brassica digenomic species in the U's triangle, i.e., B. napus, B. juncea, and B. carinata, were detected by the probes of their parental monogenomic species, i.e., B. rapa, B. nigra, and B. oleracea. The probe for B. oleracea showed signals of B. balearica, B. cretica, B. incana, B. insularis, and B. macrocarpa, which have the C genome as B. oleracea. Eruca vesicaria DNA was detected by the probe for E. sativa, which has been classified as a subspecies of E. vescaria. DNA of leaf tissue extracted by an alkaline solution and seed DNA prepared by the NaI method can be used directly for dot-blotting. Misidentification of species was revealed in 20 accessions in the Tohoku University Brassica Seed Bank. These results indicate dot-blot hybridization to be a simple and efficient technique for identification of plant species in a gene bank. PMID:20683723

  3. Refinement of the high-resolution physical and genetic map of Rhodobacter capsulatus and genome surveys using blots of the cosmid encyclopedia.

    PubMed Central

    Fonstein, M; Koshy, E G; Nikolskaya, T; Mourachov, P; Haselkorn, R

    1995-01-01

    Cosmids from a library containing Rhodobacter capsulatus DNA fragments were previously ordered in two contigs: one corresponding to the chromosome and one to a 134 kb plasmid. This map contained 40 regions connected only by colony hybridization. To confirm the linkage and correct the map, the actual sizes of the overlaps were determined by blot-hybridization with Rhodobacter chromosomal DNA and by mapping of additional cosmids. Several revisions of the earlier map include single cosmid shifts and inversions. One additional gap in a cosmid contig was also found, raising the possibility that the chromosome is not a contiguous circle. About 2500 additional EcoRI,BamHI and HindIII restriction sites were added to the 560 EcoRV sites previously mapped onto the Rhodobacter chromosome, increasing the resolution of the physical map to the size of individual genes. Twenty-five new markers were located on the genetic map. The 48 markers now mapped represent nearly 300 genes and ORFs cloned from different species of Rhodobacter. The orientation of transcription of the four rrn operons was established using 16S rRNA- and 23S rRNA-specific probes and digestion with the rare-cutting enzyme, CeuI. Gel blots of 192 cosmids of the miniset of R.capsulatus digested with EcoRV were prepared. Such a hybridization template represents the whole genome cut into 560 DNA fragments varying in size from 0.4 to 25 kb. This template was used for high-resolution mapping of single genes, analysis of total genomic DNAs from related Rhodobacter strains and differentially expressed RNAs. Images PMID:7737133

  4. A western blot assay to measure cyclin dependent kinase activity in cells or in vitro without the use of radioisotopes.

    PubMed

    Lewis, Cody W; Taylor, Ryan G; Kubara, Philip M; Marshall, Kris; Meijer, Laurent; Golsteyn, Roy M

    2013-09-17

    We developed a quantitative method to measure the activity of cyclin-dependent kinases (Cdks) by western blotting, without radioisotopes. We prepared a recombinant protein substrate based upon the natural Cdk1 substrate, PP1Cα. By combining this substrate in a western blot method using fluorochrome based antibodies and phospho-imager analysis, we measured the Km of ATP binding to Cdk1 to be 3.5 μM. We then measured Cdk1 activity in cell extracts from interphase or mitotic cells, and demonstrated that previously identified Cdk inhibitors could be detected by this assay. Our data show that we have a safe, reliable assay to identify Cdk1 inhibitors and measure Cdk1 activity. PMID:23954627

  5. Immunological Diagnosis of Human Cystic Echinococcosis: Utility of Discriminant Analysis Applied to the Enzyme-Linked Immunoelectrotransfer Blot

    PubMed Central

    Gadea, I.; Ayala, G.; Diago, M. T.; Cuñat, A.; de Lomas, J. García

    1999-01-01

    An enzyme-linked immunoelectrotransfer blot for the diagnosis of human hydatid disease was performed, and the different antibody responses were analyzed by a discriminant analysis. This multivariate technique gave us, first, a selection of the most important responses against Echinococcus granulosus infection and, second, a procedure for the classification of patients into two groups: patients with hydatid disease and patients without a history of hydatid disease. This method was applied to 67 patients, 25 with active hydatid cysts (24 hepatic and 1 pulmonary) and 42 without a history of hydatid disease and was compared with the results obtained by conventional serology: indirect hemagglutination, latex particle agglutination, and basophil degranulation. An immunoelectrotransfer blot coupled to a discriminant analysis was more sensitive than conventional serological diagnosis and detected 100% of patients with an active hepatic hydatid cyst with a specificity of 100%. This method, however, failed to detect an uncomplicated hyaline pulmonary hydatid cyst. PMID:10391851

  6. Western blot (immunoblot) assay of small, round-structured virus associated with an acute gastroenteritis outbreak in Tokyo.

    PubMed Central

    Hayashi, Y; Ando, T; Utagawa, E; Sekine, S; Okada, S; Yabuuchi, K; Miki, T; Ohashi, M

    1989-01-01

    Small, round-structured virus (SRSV) was detected in a stool specimen of a patient during an acute gastroenteritis outbreak in Tokyo and was tentatively named SRSV-9. SRSV-9 was purified by sucrose velocity gradient centrifugation after CsCl density gradient centrifugation. The buoyant density of SRSV-9 appeared to be 1.36 g/ml in CsCl. A Western blot (immunoblot) assay using the biotin-avidin system revealed that SRSV-9 was antigenically related to the Hawaii agent but distinct from the Norwalk agent and contained a single major structural protein with a molecular size of 63.0 +/- 0.6 kilodaltons. The prevalence of SRSV-9 infection in Tokyo was surveyed by the Western blot antibody assay by using a crude virus preparation as the antigen. Seroconversion was observed in 56.5% of the patients involved in the outbreaks from which SRSV was detected by electron microscopy. Images PMID:2504773

  7. Densitometric analysis of Western blot (immunoblot) assays for human immunodeficiency virus antibodies and correlation with clinical status.

    PubMed Central

    Schmidt, G; Amiraian, K; Frey, H; Stevens, R W; Berns, D S

    1987-01-01

    Western blot assays for antibodies directed against components of human immunodeficiency virus (HIV) associated with acquired immunodeficiency syndrome (AIDS) were examined with a densitometer and integrator. Antibody responses to seven HIV proteins were determined from the areas under the peaks of bands on blots from 430 seropositive individuals. Antibody responses corresponded qualitatively and quantitatively with clinical status. The Western blot assays examined were done on single specimens from individuals in one of four clinical states: asymptomatic with no risk factor identified, asymptomatic with risk factor(s) identified, AIDS-related complex, and AIDS. The ratios of gp41 antibody to p24 antibody and of gp41 antibody to total HIV antibodies increased, and the number of total HIV antibodies decreased progressively in these populations. Parameters were assigned to characterize the typical response found in AIDS: gp41 antibody/p24 antibody ratio, greater than or equal to 2.0; gp41 antibody/total HIV antibodies ratio, greater than or equal to 0.30; and number of total HIV antibodies, less than or equal to 25.0 signal units. Parameter match increased with progression of clinical status. These parameters were applied in a brief follow-up study of 34 HIV-infected asymptomatic individuals who developed AIDS-related complex or AIDS. Initial specimens showed a stronger correlation than our population data base had predicted, suggesting that the parameters have prognostic value. Densitometric analysis of antibody responses on Western blot assays of single or serial specimens should prove useful to physicians in staging and monitoring HIV-infected individuals and in predicting which individuals will progress to AIDS. Images PMID:2444624

  8. Cross antigenicity of immunodominant polypeptides of somatic antigen of Oesophagostomum columbianum with other helminths by western blotting

    PubMed Central

    Dalal, Sunita; Prasad, Arvind; Nasir, Abdul; Saini, Vijesh Kumar

    2015-01-01

    Aim: Oesophagostomum columbianum in small ruminants in India is found as mixed infection commonly in sheep and goat. Haemonchus contortus, an abomasal nematode is found as concurrent infection with it. Eggs of Haemonchus and O. columbianum cannot be easily distinguished. Diagnosis of O. columbianum may only be possible if a non-cross antigenic polypeptide was available for immunodiagnosis. Materials and Methods: Somatic antigen (SoAg) of O. columbianum was fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunodominant polypeptides were identified by western blotting with homologous hyperimmune serum (HIS) and experimental sera of sheep or goat infected with other helminths. Results: SoAg of O. columbianum was immunoaffinity purified. Sharp polypeptide bands of 130, 72 and 68 KDa were observed along with several faint bands of lower molecular weight. Western blot of purified SoAg of O. columbianum with homologous HIS showed reaction with all the protein bands of 17, 28, 30, 32, 35, 38, 50, 68, 100, 130, 150, and 170 kDa. For identification of non-cross antigenic polypeptide, immunoaffinity purified SoAg of O. columbianum was reacted to heterologous HIS against H. contortus, Paramphistomum epiclitum, and Fasciola gigantica in western blotting utilizing completely dry method (i-blot). Among high molecular weight polypeptides 100 and 150 kDa were non-cross antigenic and among low molecular weight except 50 kDa polypeptide, 17, 30, 32, 35, and 38 kDa of O. columbianum were not cross antigenic with other helminths. Conclusions: Hence, polypeptides of 17, 30, 32, 35 and 38 kDa as well as 100 and 150 kDa polypeptides of O. columbianum may be exploited for immunodiagnosis of the infection in sheep and goat with extensive studies on cross antigenicity. PMID:27047030

  9. Use of Phaseolus vulgaris leukoagglutinating lectin in histochemical and blotting techniques: a comparison of digoxigenin- and biotin-labelled lectins.

    PubMed

    Li, W P; Zuber, C; Roth, J

    1993-11-01

    An increase in the number of beta 1,6 branches of the trimannosyl core of asparagine-linked oligosaccharides has been shown to be directly correlated with the metastatic potential of cultured tumour cells. The Phaseolus vulgaris leukoagglutinating lectin (PHA-L) binds to beta 1,6 branches of tri- and tetra-antennary oligosaccharides. We have applied digoxigenin- and biotin-conjugated PHA-L to establish a non-radioactive detection system for beta 1,6 branches, which can be used in lectin blotting as well as light and electron microscopic cytochemistry. For this purpose the HCT116 human colon carcinoma cell line and colon carcinoma tissue were investigated. Digoxigenin-conjugated PHA-L in conjunction with alkaline phosphatase-conjugated anti-digoxigenin antibodies was superior to biotin-conjugated PHA-L in lectin blotting with respect to sensitivity and specificity. Similarly, the digoxigenin conjugated PHA-L in conjunction with gold-labelled anti-digoxigenin antibodies resulted in more intense specific staining and lower background compared to biotin-conjugated PHA-L visualized with a streptavidin immunogold complex. The specificity of lectin binding in blotting and cytochemical studies was demonstrated by the absence of staining when the lectin was omitted or preabsorbed with glycoprotein, and following pretreatment of the cellular homogenates or tissue sections by N-glycosidase F. Our results demonstrate that digoxigenin-conjugated PHA-L provides high sensitivity and specificity for histochemical and blotting techniques and is amenable for quantification. The technique should have applications in tumour research. PMID:7508428

  10. Western blot can distinguish natural and acquired antibodies to Mycoplasma agassizii in the desert tortoise (Gopherus agassizii).

    PubMed

    Hunter, Kenneth W; Dupré, Sally A; Sharp, Tiffanny; Sandmeier, Franziska C; Tracy, C Richard

    2008-12-01

    Mycoplasma agassizi has been identified as a cause of upper respiratory tract disease (URTD) in the threatened Mojave population of the desert tortoise (Gopherus agassizii), and anti-M. agassizii antibodies have been found by ELISA in as many as 15% of these animals across their geographic range. Here we report that a cohort of 16 egg-reared desert tortoises never exposed to M. agassizii had ELISA antibody titers to this organism that overlapped with titers obtained from some M. agassizii-infected tortoises. These natural antibodies were predominantly of the IgM class. Western blots of plasma from these non-infected tortoises produced a characteristic banding pattern against M. agassizii antigens. A group of 38 wild-caught desert tortoises was tested by ELISA, and although some of these tortoises had antibody titers significantly higher than the non-infected tortoises, there was considerable overlap at the lower titer levels. However, Western blot analysis revealed distinct banding patterns that could readily distinguish between the non-infected tortoises and tortoises with acquired antibodies, regardless of ELISA antibody titers. We conclude that desert tortoises have natural antibodies to M. agassizii that can compromise the determination of infection status by ELISA. However, the Western blot technique can distinguish between natural and acquired antibody patterns and can be used to confirm the diagnosis of M. agassizii infections in the desert tortoise. PMID:18708096

  11. Identification of Novel Laminin- and Fibronectin-binding Proteins by Far-Western Blot: Capturing the Adhesins of Streptococcus suis Type 2

    PubMed Central

    Li, Quan; Liu, Hanze; Du, Dechao; Yu, Yanfei; Ma, Caifeng; Jiao, Fangfang; Yao, Huochun; Lu, Chengping; Zhang, Wei

    2015-01-01

    Bacterial cell wall (CW) and extracellular (EC) proteins are often involved in interactions with extracellular matrix (ECM) proteins such as laminin (LN) and fibronectin (FN), which play important roles in adhesion and invasion. In this study, an efficient method combining proteomic analysis and Far-Western blotting assays was developed to screen directly for bacterial surface proteins with LN- and FN-binding capacity. With this approach, fifteen potential LN-binding proteins and five potential FN-binding proteins were identified from Streptococcus suis serotype 2 (SS2) CW and EC proteins. Nine newly identified proteins, including oligopeptide-binding protein OppA precursor (OppA), elongation factor Tu (EF-Tu), enolase, lactate dehydrogenase (LDH), fructose-bisphosphate aldolase (FBA), 3-ketoacyl-ACP reductase (KAR), Gly ceraldehyde-3-phosphate dehydrogenase (GAPDH), Inosine 5′-monophosphate dehydrogenase (IMPDH), and amino acid ABC transporter permease (ABC) were cloned, expressed, purified and further confirmed by Far-Western blotting and ELISA. Five proteins (OppA, EF-Tu, enolase, LDH, and FBA) exhibited specifically binding activity to both human LN and human FN. Furthermore, seven important recombinant proteins were selected and identified to have the ability to bind Hep-2 cells by the indirect immunofluorescent assay. In addition, four recombinant proteins, and their corresponding polyclonal antibodies, were observed to decrease SS2 adhesion to Hep-2 cells, which indicates that these proteins contribute to the adherence of SS2 to host cell surface. Collectively, these results show that the approach described here represents a useful tool for investigating the host-pathogen interactions. PMID:26636044

  12. Genetics Home Reference: Northern epilepsy

    MedlinePlus

    ... Understand Genetics Home Health Conditions Northern epilepsy Northern epilepsy Enable Javascript to view the expand/collapse boxes. Download PDF Open All Close All Description Northern epilepsy is a genetic condition that causes recurrent seizures ( ...

  13. Western blot patterns of serum autoantibodies against optic nerve antigens in dogs with goniodysgenesis-related glaucoma

    PubMed Central

    Pumphrey, Stephanie A.; Pizzirani, Stefano; Pirie, Christopher G.; Anwer, M. Sawkat; Logvinenko, Tanya

    2014-01-01

    Objective To investigate whether differences existed between clinically normal dogs and dogs with goniodysgenesis-related glaucoma (GDRG) in serum autoantibodies against optic nerve antigens. Animals 16 dogs with GDRG, 17 healthy dogs with unremarkable pectinate ligament and iridocorneal angle morphology, and 13 euthanized dogs with no major ocular abnormalities or underlying diseases. Procedures Western blotting was performed with optic nerve extracts from the euthanized dogs as an antigen source and serum from clinically normal dogs and dogs with GDRG as a primary antibody (autoantibody) source. Blots were evaluated for presence and density of bands. Results Multiple bands were identified on western blots from all dogs with GDRG and all clinically normal dogs, with a high degree of variability among individual dogs. Dogs with GDRG were significantly more likely than healthy dogs to have bands present at 38, 40, and 68 kDa. Dogs with GDRG had significant increases in autoreactivity at 40 and 53 kDa and a significant decrease in autoreactivity at 48 kDa. Conclusions and Clinical Relevance Significant differences in serum autoantibodies against optic nerve antigens were found in dogs with versus without GDRG. Although it remains unclear whether these differences were part of the pathogenesis of disease or were sequelae to glaucomatous changes, these findings provide support for the hypothesis that immune-mediated mechanisms play a role in the development or progression of GDRG. However, the high degree of variability among individual dogs and the considerable overlap between groups suggest that the clinical usefulness of this technique for distinguishing dogs with GDRG from clinically normal dogs is likely limited. PMID:23531071

  14. Diagnosis of enteric fever caused by Salmonella spp. in Vietnam by a monoclonal antibody-based dot-blot ELISA.

    PubMed

    Nguyen, N Q; Tapchaisri, P; Chongsa-nguan, M; Cao, V V; Doan, T T; Sakolvaree, Y; Srimanote, P; Chaicumpa, W

    1997-12-01

    Enteric fever caused by Salmonella spp. is prevalent in Vietnam. None of the currently available diagnostic methods meets the ideal criteria on rapidity, simplicity, sensitivity, specificity, cost-effectiveness and practicality for developing areas. In this study, a recently developed monoclonal antibody-based dot-blot ELISA was used in comparison with the hemoculture method and the classical Widal test for diagnosis of salmonellosis in 171 Vietnamese patients presenting with clinical features of enteric fever. Urine samples of 50 healthy counterparts were used as negative controls. Salmonella spp. were isolated from 77 of 171 patients (45%) while 98 and 111 patients were positive by dot-blot ELISA and Widal test, respectively. The diagnostic sensitivity, specificity, accuracy, positive predictive value and negative predictive value of the ELISA performed on three serial urine samples collected at 2 hour intervals of the 171 patients were 92.2%, 71.3%, 80.7%, 72.4% and 91.8%, respectively when compared with the culture method. The Widal test performed on acute and convalescence serum samples showed 87.0%, 46.8%, 68.4%, 60.4% and 83.3% diagnostic sensitivity, specificity, accuracy, and positive and negative predictive values, respectively when compared with the bacterial culture method. Kappa coefficience revealed very good agreement beyond chance between the MAb-based ELISA and the culture method. The ELISA was not reactive when tested on urine samples of 50 healthy individuals which indicates 100% specificity. The Salmonella antigenuria of the patients as detected by ELISA lasted 10.3+/-3.9 days after initiating antibiotic treatment. The MAb-based dot-blot ELISA is easy to perform. It is rapid, sensitive, specific, inexpensive, and non-invasive and does not require equipment, thus is suitable for developing areas. It can detect acute/recent infection and can be used for evaluation of the efficacy of the treatment. PMID:9579614

  15. Rapid and correct identification of intestinal Bacteroides spp. with chromosomal DNA probes by whole-cell dot blot hybridization

    SciTech Connect

    Morotomi, M.; Ohno, T.; Mutai, M.

    1988-05-01

    A dot blot hybridization procedure with /sup 32/P-labeled whole chromosomal DNA of the type strains as probes was developed as a rapid and simple method for identification of intestinal Bacteroides species. Bacterial cells were fixed onto membrane filters by slight suction, treated with 0.5 N NaOH, and hybridized with these probes. Of 65 Bacteroides strains isolated from 19 human fecal specimens, which were identified as B. fragilis, B. thetaiotaomicron, B. ovatus, B. caccae, B. uniformis, B. stercoris, B. vulgatus, B. distasonis, and B. merdae by conventional phenotypic characterization, 62 (95%) were correctly identified with this hybridization procedure.

  16. Immunohistochemical and Western Blotting Analyses of Ganoine in the Ganoid Scales of Lepisosteus oculatus: an Actinopterygian Fish.

    PubMed

    Sasagawa, Ichiro; Oka, Shunya; Mikami, Masato; Yokosuka, Hiroyuki; Ishiyama, Mikio; Imai, Akane; Shimokawa, Hitoyata; Uchida, Takashi

    2016-05-01

    In order to compare its characteristics with those of jaw tooth collar enamel, normally developing and experimentally regenerating ganoine from ganoid scales of Lepisosteus oculatus (spotted gar), an actinopterygian fish species, was examined by Western blotting and immunohistochemistry. Amelogenin, a major enamel matrix protein (EMP), is widely found from sarcopterygian fish to mammals. Therefore, we used antimammalian amelogenin antibodies and antisera: an antibody against bovine amelogenin; antiserum against porcine amelogenin; and region-specific antibodies or antiserum against the C-terminus, middle region, or N-terminus of porcine amelogenin in this study. Positive immunoreactivity with the antibody against bovine amelogenin, antiserum against porcine amelogenin, and the middle and C-terminal region-specific antibodies was detected in both normally developing and regenerating ganoine matrix, as well as in granules found within inner ganoine epithelial cells. These immunohistochemical analyses indicated that the Lepisosteus ganoine matrix contains EMP-like proteins with epitopes similar to mammalian amelogenins. In Western blotting analyses of regenerating ganoid scales with the antibovine amelogenin antibody, two protein bands with molecular weights of approximately 78 and 65 kDa were detected, which were similar to those found in Lepisosteus tooth enamel. Our study suggests that in Lepisosteus, EMP-like proteins in the ganoine matrix corresponded to those in tooth enamel. However, it was revealed that the 78 and 65 kDa EMP-like proteins were different from 27 kDa bovine amelogenin. PMID:27139791

  17. Mutation analysis of fragile X syndrome by Southern blot, radioactive PCR, silver-stained polyacrylamide gel and DIG DNA

    SciTech Connect

    Lee, Sook-Hwan; Kim, Un-Kyung; Chung-Woong, M.S.

    1994-09-01

    Fragile X syndrome is the most common inherited form of mental retardation. In fragile X syndrome, the underlying mutation is caused by an expansion of the CTG triplet in the 5{prime} untranslated region of the FMR-1 gene located at Xq27.3 and diagnosed by methylation of the associated CpG island. This disorder becomes clinically manifested when the mutation is caused by an expansion of (CGG)n reaching a threshold of about 600bp (200 repeats). The number of inserted repeats increases through the generation. We have analyzed fragile X syndrome by 4 different methods: Southern blot, radioactive PCR, polyacrylamide gel and DIG DNA labeling/detection techniques. Southern blot and DIG DNA labeling/detection by double DNA digestion with EcoRI and EagI reveals both the presence of the mutation and the methylation status. Radioactive PCR and silver-stained polyacrylamide gel is a rapid and sensitive technique to define the unaffected carriers and NTMs, but it is difficult to amplify such a highly GC-rich sequence. Further testing in other fragile X patients is currently in progress.

  18. Identification of Glioblastoma Phosphotyrosine-Containing Proteins with Two-Dimensional Western Blotting and Tandem Mass Spectrometry

    PubMed Central

    Guo, Tianyao; Wang, Xiaowei; Li, Maoyu; Yang, Haiyan; Li, Ling; Peng, Fang

    2015-01-01

    To investigate the presence of, and the potential biological roles of, protein tyrosine phosphorylation in the glioblastoma pathogenesis, two-dimensional gel electrophoresis- (2DGE-) based Western blotting coupled with liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) analysis was used to detect and identify the phosphotyrosine immunoreaction-positive proteins in a glioblastoma tissue. MS/MS and Mascot analyses were used to determine the phosphotyrosine sites of each phosphopeptide. Protein domain and motif analysis and systems pathway analysis were used to determine the protein domains/motifs that contained phosphotyrosine residue and signal pathway networks to clarify the potential biological functions of protein tyrosine phosphorylation. A total of 24 phosphotyrosine-containing proteins were identified. Each phosphotyrosine-containing protein contained at least one tyrosine kinase phosphorylation motif and a certain structural and functional domains. Those phosphotyrosine-containing proteins were involved in the multiple signal pathway systems such as oxidative stress, stress response, and cell migration. Those data show 2DGE-based Western blotting, MS/MS, and bioinformatics are a set of effective approaches to detect and identify glioblastoma tyrosine-phosphorylated proteome and to effectively rationalize the biological roles of tyrosine phosphorylation in the glioblastoma biological systems. It provides novel insights regarding tyrosine phosphorylation and its potential role in the molecular mechanism of a glioblastoma. PMID:26090378

  19. Simple and Sensitive Detection of HBsAg by Using a Quantum Dots Nanobeads Based Dot-Blot Immunoassay

    PubMed Central

    Zhang, Pengfei; Lu, Huiqi; Chen, Jia; Han, Huanxing; Ma, Wei

    2014-01-01

    Simple and sensitive detection of infectious disease at an affordable cost is urgently needed in developing nations. In this regard, the dot blot immunoassay has been used as a common protein detection method for detection of disease markers. However, the traditional signal reporting systems, such as those using enzymes or gold nanoparticles lack sensitivity and thus restrict the application of these methods for disease detection. In this study, we report a simple and sensitive detection method for the detection of infectious disease markers that couples the dot-blot immunoassay with quantum dots nanobeads (QDNBs) as a reporter. First, the QDNBs were prepared by an oil-in-water emulsion-evaporation technique. Because of the encapsulation of several QDs in one particle, the fluorescent signal of reporter can be amplified with QDNBs in a one-step test and be read using a UV lamp obviating the need for complicated instruments. Detection of disease-associated markers in complex mixture is possible, which demonstrates the potential of developing QDNBs into a sensitive diagnostic kit. PMID:24505238

  20. Simple and sensitive detection of HBsAg by using a quantum dots nanobeads based dot-blot immunoassay.

    PubMed

    Zhang, Pengfei; Lu, Huiqi; Chen, Jia; Han, Huanxing; Ma, Wei

    2014-01-01

    Simple and sensitive detection of infectious disease at an affordable cost is urgently needed in developing nations. In this regard, the dot blot immunoassay has been used as a common protein detection method for detection of disease markers. However, the traditional signal reporting systems, such as those using enzymes or gold nanoparticles lack sensitivity and thus restrict the application of these methods for disease detection. In this study, we report a simple and sensitive detection method for the detection of infectious disease markers that couples the dot-blot immunoassay with quantum dots nanobeads (QDNBs) as a reporter. First, the QDNBs were prepared by an oil-in-water emulsion-evaporation technique. Because of the encapsulation of several QDs in one particle, the fluorescent signal of reporter can be amplified with QDNBs in a one-step test and be read using a UV lamp obviating the need for complicated instruments. Detection of disease-associated markers in complex mixture is possible, which demonstrates the potential of developing QDNBs into a sensitive diagnostic kit. PMID:24505238

  1. Tissue blot immunoassay and direct RT-PCR of cucumoviruses and potyviruses from the same NitroPure nitrocellulose membrane.

    PubMed

    Chang, Peta-Gaye S; McLaughlin, Wayne A; Tolin, Sue A

    2011-02-01

    A method is described for using Nitropure nitrocellulose (NPN) membranes as an effective solid matrix for retrieval of template RNA of three potyviruses, Tobacco etch virus, Soybean mosaic virus and Turnip mosaic virus, and two cucumoviruses, Cucumber mosaic virus and Peanut stunt virus. These NPN membranes were also used for tissue blot immunosorbent assays (TBIAs) to identify and detect plant viruses. For RNA detection, discs from dried membranes blotted with infected tissue were minimally cleaned with Triton X-100 and placed directly into reverse transcription (RT) reactions to initiate cDNA synthesis. Aliquots of cDNA plus primers specific for coat protein produced PCR amplicons of expected sizes for each of the viruses. Intensity of PCR-amplified bands from cDNA transcribed from both non-processed and TBIA-processed NPN membranes was comparable to those using FTA Card protocols. Direct sequencing of PCR products yielded high quality runs enabling identification to species. NPN membranes retained immunologically detectable virus particles, as well as intact template viral RNA, for more than a year at room temperature. The quantity of amplification product decreased after several months of storage, but could be increased by increasing the number of PCR cycles. PMID:21126542

  2. Phylogenetic distribution of apolipoproteins A-I and E in vertebrates as determined by Western blot analysis.

    PubMed

    Duggan, A E; Callard, I P

    2001-08-01

    A putative apolipoprotein E (apoE) has been identified in the HDL and VHDL fractions of the turtle. This observation is of particular interest considering apoE has been reported absent in the domestic hen (Hermier et al., '95; Biochim Biophys Acta: 105-118, 1995) and thus presumed absent in nonmammalian vertebrates altogether. As a result, partial amino acid sequencing of this protein was performed and revealed that one fragment shared 41% sequence identity to human apoE. Western blot analysis using antisera to apoE demonstrated cross-reactivity to a 34-kDa protein (putative apoE) in turtle plasma. Further investigation using anti-apoE antibody in Western blot analysis detected immunoreactive apoE in the plasma of lamprey, spiny dogfish, skate, and alligator, but not in flounder, newt or python; its absence in several species of birds was confirmed. Using anti-apoA-I antibody, apoA-I was detected in all vertebrate groups except a representative teleost (flounder). Apo-A-I antibody cross-reacted weakly with some putative apoE proteins (chicken, spiny dogfish and skate) and the reverse was true for anti-apoE, which cross-reacted with putative apoA-I in birds, reptiles, and elasmobranchs, confirming the molecular similarity and phylogenetic relatedness of these two proteins. PMID:11479905

  3. Transcription factor proteomics: identification by a novel gel mobility shift-three-dimensional electrophoresis method coupled with southwestern blot and high-performance liquid chromatography-electrospray-mass spectrometry analysis.

    PubMed

    Jiang, Daifeng; Jia, Yinshan; Jarrett, Harry W

    2011-09-28

    Transcription factor (TF) purification and identification is an important step in elucidating gene regulatory mechanisms. In this study, we present two new electrophoretic mobility shift assay (EMSA)-based multi-dimensional electrophoresis approaches to isolate and characterize TFs, using detection with either southwestern or western blotting and HPLC-nanoESI-MS/MS analysis for identification. These new techniques involve several major steps. First, EMSA is performed with agents that diminish non-specific DNA-binding and the DNA-protein complex is separated by native PAGE gel. The gel is then electrotransferred to PVDF membrane and visualized by autoradiography. Next, the DNA-protein complex, which has been transferred onto the blot, is extracted using a detergent-containing elution buffer. Following detergent removal, concentrated extract is separated by SDS-PAGE (EMSA-2DE), followed by in-gel trypsin digestion and HPLC-nanoESI-MS/MS analysis, or the concentrated extract is separated by two-dimensional gel electrophoresis (EMSA-3DE), followed by southwestern or western blot analysis to localize DNA binding proteins on blot which are further identified by on-blot trypsin digestion and HPLC-nanoESI-MS/MS analysis. Finally, the identified DNA binding proteins are further validated by EMSA-immunoblotting or EMSA antibody supershift assay. This approach is used to purify and identify GFP-C/EBP fusion protein from bacterial crude extract, as well as purifying AP1 and CEBP DNA binding proteins from a human embryonic kidney cell line (HEK293) nuclear extract. AP1 components, c-Jun, Jun-D, c-Fos, CREB, ATF1 and ATF2 were successfully identified from 1.5 mg of nuclear extract (equivalent to 3×10(7) HEK293 cells) with AP1 binding activity of 750 fmol. In conclusion, this new strategy of combining EMSA with additional dimensions of electrophoresis and using southwestern blotting for detection proves to be a valuable approach in the identification of transcriptional complexes

  4. 7 CFR 457.142 - Northern potato crop insurance provisions.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 7 CFR part 400, subpart T, and pay an additional premium, you may increase your prevented planting... 7 Agriculture 6 2011-01-01 2011-01-01 false Northern potato crop insurance provisions. 457.142... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.142 Northern...

  5. 7 CFR 457.142 - Northern potato crop insurance provisions.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 7 CFR part 400, subpart T, and pay an additional premium, you may increase your prevented planting... 7 Agriculture 6 2014-01-01 2014-01-01 false Northern potato crop insurance provisions. 457.142... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.142 Northern...

  6. 7 CFR 457.142 - Northern potato crop insurance provisions.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 7 CFR part 400, subpart T, and pay an additional premium, you may increase your prevented planting... 7 Agriculture 6 2013-01-01 2013-01-01 false Northern potato crop insurance provisions. 457.142... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.142 Northern...

  7. 7 CFR 457.142 - Northern potato crop insurance provisions.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 7 CFR part 400, subpart T, and pay an additional premium, you may increase your prevented planting... 7 Agriculture 6 2012-01-01 2012-01-01 false Northern potato crop insurance provisions. 457.142... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.142 Northern...

  8. 7 CFR 457.142 - Northern potato crop insurance provisions.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 7 CFR part 400, subpart T, and pay an additional premium, you may increase your prevented planting... 7 Agriculture 6 2010-01-01 2010-01-01 false Northern potato crop insurance provisions. 457.142... INSURANCE CORPORATION, DEPARTMENT OF AGRICULTURE COMMON CROP INSURANCE REGULATIONS § 457.142 Northern...

  9. Tornadoes Strike Northern Wisconsin

    NASA Technical Reports Server (NTRS)

    2007-01-01

    A series of tornadoes ripped through the Upper Midwest region of the United States in the evening of June 7, 2007. At least five different tornadoes touched down in Wisconsin, according to the Associated Press, one of which tore through the Bear Paw Resort in northern Wisconsin. Despite dropping as much as fifteen centimeters (six inches) of rain in some places and baseball-size hail in others, authorities were reporting no deaths attributable to the storm system, and only a smattering of injuries, but considerable property damage in some areas. When the MODIS instrument on NASA's Terra satellite observed the area on June 9, 2007, the track torn through the woods by one of the tornadoes stands out quite clearly. This photo-like image uses data collected by MODIS in the normal human vision range to give a familiar natural-looking appearance. The landscape is largely a checkerboard of farms, towns, roads, and cities. The pale land is predominantly farmland where crops have not fully grown in yet. Dark blue shows the winding path of rivers and lakes dotting the landscape. The large blue lake on the east (right) side of the image is Lake Michigan. Towns and cities, including the city of Green Bay, are gray. To the north side, farmland gives way to dark green as land use shifts from agriculture to the Menominee Indian Reservation and Nicolet National Forest. The diagonal slash through the dark green forested land shows the tornado track. Bare land was revealed where the tornado tore down trees or stripped vegetation off the branches. The high-resolution image provided above is at MODIS' full spatial resolution (level of detail) of 250 meters per pixel. The MODIS Rapid Response System provides this image at additional resolutions.

  10. Direct assay of thymidine kinase bound to ion-exchange paper for dot spotting and enzyme blotting analysis

    SciTech Connect

    van den Berg, K.J.

    1986-05-15

    The direct assay of thymidine kinase (Tk) bound to ion-exchange paper was investigated as a means to further simplify the analytical procedure. Thymidine kinase bound firmly and quantitatively to ion-exchange paper at near neutral pH. The enzymatic properties of Tk did not change while bound to the ion-exchange paper. The amount of phosphorylated /sup 12//sub 5/IdU or /sup 125/IdC formed on ion-exchange paper was proportional to the amount of applied Tk. Enzymatic activity could be determined visually by autoradiography or by gamma counting. This method was relatively independent of the protein concentration or volume of the sample and which allows the assay from dilute solutions. A simplified dot spot method that can be used for the assay of thymidine kinase activity in cell extracts is described. Thymidine kinase could also be visualized after electrophoresis and blotting on ion-exchange paper.

  11. [THE SCREENING OF DIAGNOSTIC POTENTIAL OF NATIVE PROTEIN FRACTIONS OF MYCOBACTERIUM TUBERCULOSIS USING TECHNIQUE OF IMMUNE BLOTTING].

    PubMed

    Tsibulkin, A P; Khaertinova, I M; Urazov, N G; Khaertinov, K S

    2016-02-01

    The technique of immune blotting was used to analyze surface proteins obtained from cells M Tuberculosis exposed to partialmode of delipidization. At that, there were applied serums of patients with tuberculosis of lungs; HIV agents and patients with concomitant infection tuberculosis-AIDS and also HIV-negative patients without clinical signs of disease of lungs and with chronic diseases of lungs of other etiology The fractions oflow-molecular antigens with molecular mass 6.5-30 kilodaltons became diagnostically significant. To this fraction of antigens reacted serums of all patients with tuberculosis of lungs and serums of 91% of patients with concomitant tuberculosis-AIDS infection. The antigens of protein fractions with high (70-100 kilodaltons) and interim (30-69 kilodaltons) molecular mass became diagnostically insignificant. PMID:27455562

  12. Rabbit antisera against three different bacteria which can induce reactive arthritis: analysis by ELISA, immunoprecipitation and Western Blot.

    PubMed Central

    Ogasawara, M; Kobayashi, S; Hill, J L; Kono, D H; Yu, D T

    1985-01-01

    Three strains of bacteria which induce reactive arthritis were collected: a Shigella flexneri, designated 7060; another Sh. flexneri, designated 316; and a Yersinia enterocolitica of serotype 03. Rabbit antisera were generated against each of them to test for the extent and nature of cross-reactivity among these strains. When analysed by the ELISA technique, antisera against 7060 and 316 showed strong cross-reactivity with Y. enterocolitica. In contrast, the reaction of antisera prepared against putatively non-arthritis-causing bacteria reacted several-folds less. Using immunoprecipitation, sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western Blot procedures, a 92,000 MW cross-reactive antigen on the Yersinia was identified. The antigen was present on the outer membranes of the Y. enterocolitica, and enzyme digestion experiments showed that this antigen was protein in nature. Images Figure 1 Figure 2 Figure 3 Figure 4 PMID:3884493

  13. Identification of Xanthomonas fragariae, Xanthomonas axonopodis pv. phaseoli, and Xanthomonas fuscans subsp. fuscans with Novel Markers and Using a Dot Blot Platform Coupled with Automatic Data Analysis ▿ †

    PubMed Central

    Albuquerque, Pedro; Caridade, Cristina M. R.; Marcal, Andre R. S.; Cruz, Joana; Cruz, Leonor; Santos, Catarina L.; Mendes, Marta V.; Tavares, Fernando

    2011-01-01

    Phytosanitary regulations and the provision of plant health certificates still rely mainly on long and laborious culture-based methods of diagnosis, which are frequently inconclusive. DNA-based methods of detection can circumvent many of the limitations of currently used screening methods, allowing a fast and accurate monitoring of samples. The genus Xanthomonas includes 13 phytopathogenic quarantine organisms for which improved methods of diagnosis are needed. In this work, we propose 21 new Xanthomonas-specific molecular markers, within loci coding for Xanthomonas-specific protein domains, useful for DNA-based methods of identification of xanthomonads. The specificity of these markers was assessed by a dot blot hybridization array using 23 non-Xanthomonas species, mostly soil dwelling and/or phytopathogens for the same host plants. In addition, the validation of these markers on 15 Xanthomonas spp. suggested species-specific hybridization patterns, which allowed discrimination among the different Xanthomonas species. Having in mind that DNA-based methods of diagnosis are particularly hampered for unsequenced species, namely, Xanthomonas fragariae, Xanthomonas axonopodis pv. phaseoli, and Xanthomonas fuscans subsp. fuscans, for which comparative genomics tools to search for DNA signatures are not yet applicable, emphasis was given to the selection of informative markers able to identify X. fragariae, X. axonopodis pv. phaseoli, and X. fuscans subsp. fuscans strains. In order to avoid inconsistencies due to operator-dependent interpretation of dot blot data, an image-processing algorithm was developed to analyze automatically the dot blot patterns. Ultimately, the proposed markers and the dot blot platform, coupled with automatic data analyses, have the potential to foster a thorough monitoring of phytopathogenic xanthomonads. PMID:21705524

  14. Serologic immunoreactivity to Neospora caninum antigens in dogs determined by indirect immunofluorescence, western blotting and dot-ELISA.

    PubMed

    Pinheiro, A M; Costa, M F; Paule, B; Vale, V; Ribeiro, M; Nascimento, I; Schaer, R E; Almeida, M A O; Meyer, R; Freire, S M

    2005-06-10

    Neospora caninum, is a coccidian protozoan known as a major cause of bovine abortion and canine neuropathies. The aim of the present study was to develop a reliable and quick test to detect antibodies to N. caninum in dog sera. Sixty-five serum samples from dogs, including 35 positive and 30 negative for N. caninum antibodies were used for standardization of the test. In parallel, immunoreactivity of the sera to Toxoplasma gondii antigens was investigated using a passive agglutination test. A dot-ELISA test, using soluble extract of N. caninum tachyzoites on nitrocellulose ester membranes, was developed and standardized. SDS-PAGE and complementary analysis of reactivity by Western blotting were used for the characterization of the immunoreactive fractions of all tested sera. The sensitivity and specificity of the dot-ELISA were 94 and 73%, respectively, compared to IFAT at a cut-off of 1:50, and 87 and 100% compared to IFAT at a cut-off of 1:25. Among the sera that tested positively for both IFAT and dot-ELISA, only 8.6% were reactive to T. gondii. The most immunoreactive fractions in Western blots were the 14-, 33-, 42- and 55 kDa bands, with percentages of 42, 60, 42 and 37%, respectively. The 60 kDa band showed a non-specific reaction in 43% of neosporosis-negative animals by both dot-ELISA and IFAT. These results indicate that the dot-ELISA using N. caninum antigen present good sensitivity and specificity, and might be used as a screening test to detect antibodies to N. caninum in dogs. PMID:15893072

  15. Imprinting mutations in Angelman syndrome detected by Southern blotting using a probe containing exon {alpha} of SNRPN

    SciTech Connect

    Beuten, J.; Sutcliffe, J.S.; Nakao, M.

    1994-09-01

    Prader-Willi syndrome (PWS) and Angelman syndrome (AS) are associated with paternal and maternal deficiencies respectively, of gene expression within human chromosome 15q11-q13, and are caused by deletion, uniparental disomy (UPD), or other mutations. The SNRPN gene maps in this region, is paternally expressed, and is a candidate gene for PWS. Southern blotting using methylation-sensitive enzymes and a genomic DNA probe from the CpG island containing exon {alpha} of the SNRPN gene reveals methylation specific for the maternal allele. In cases of the usual deletions or UPD, the probe detects absence of an unmethylated allele in PWS and absence of a methylated allele in AS. We have analyzed 21 nondeletion/nonUPD AS patients with this probe and found evidence for an imprinting mutation (absence of a methylated allele) in 3 patients. Southern blotting with methylation-sensitive enzymes using the exon {alpha} probe, like use of the PW71 probe, should detect abnormalities in all known PWS cases and in 3 of the 4 forms of AS: deletion, UPD and imprinting mutations. This analysis provides a valuable diagnostic approach for PWS and AS. In efforts to localize the imprinting mutations in AS, one patient was found with failure to inherit a dinucleotide repeat polymorphism near probe 189-1 (D15S13). Analysis of this locus in AS families and CEPH families demonstrates a polymorphism that impairs amplification and a different polymorphism involving absence of hybridization to the 189-1 probe. The functional significance, if any, of deletion of the 189-1 region is unclear.

  16. Quantitative Western ligand blotting reveals common patterns and differential features of IGFBP-fingerprints in domestic ruminant breeds and species.

    PubMed

    Wirthgen, Elisa; Höflich, Christine; Spitschak, Marion; Helmer, Carina; Brand, Bodo; Langbein, Jan; Metzger, Friedrich; Hoeflich, Andreas

    2016-02-01

    The insulin-like growth factor binding proteins (IGFBPs) are determinants of local IGF-effects and thus have an impact on growth and metabolism in vertebrate species. In farm animals, IGFBPs are associated with traits such as growth rate, body composition, milk production, or fertility. It may be assumed, that selective breeding and characteristic phenotypes of breeds are related to differential expression of IGFBPs. Therefore, the aim of the present study was to investigate the effects of selective breeding on blood IGFBP concentrations of farm animals. Breeds of the sheep, goat, and cattle species were investigated. IGFBP-3, -2, and -4 were analyzed with quantitative Western ligand blotting (qWLB), enabling comprehensive monitoring of intact IGFBPs with IGF-binding capacity. We show that in sera of all species and breeds investigated, IGFBP-3, -2, and -4 were simultaneously detectable by qWLB analysis. IGFBP-3 and the total amount of IGFBPs were significantly increased (P<0.05) in Cameroon sheep, if compared to 3 of 4 other sheep breeds, as well as in Dwarf goats versus Toggenburg and Boer goats (P<0.01). IGFBP-2 was elevated in Cameroon sheep and Boer goats, if compared to other breeds of these species (P<0.01), respectively. Holstein Friesian dairy cows had higher levels of IGFBP-4 (P<0.05), if compared to conventional crossbreeds of beef cattle. In Dwarf goats the ratio of IGFBP-3/IGFBP-2 was about 3-fold higher than in other goat breeds (P<0.001). The total IGFBP amount of Toggenburg goats was reduced (P<0.05), compared to the other goat breeds. In conclusion, our data indicate that common and specific features of IGFBP fingerprints are found in different ruminant species and breeds. Our findings may introduce quantitative Western ligand blotting as an attractive tool for biomarker development and molecular phenotyping in farm animal breeds. PMID:26597140

  17. Murine interleukin 1 receptor. Direct identification by ligand blotting and purification to homogeneity of an interleukin 1-binding glycoprotein

    SciTech Connect

    Bird, T.A.; Gearing, A.J.; Saklatvala, J.

    1988-08-25

    Functional receptors (IL1-R) for the proinflammatory cytokine interleukin 1 (IL1) were solubilized from plasma membranes of the NOB-1 subclone of murine EL4 6.1 thymoma cells using the zwitterionic detergent 3((3-cholamidopropyl)dimethylammonio)-1-propanesulfonate (CHAPS). Membrane extracts were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to nitrocellulose membranes, and ligand blotted with /sup 125/I-labeled recombinant human IL1 alpha in order to reveal proteins capable of specifically binding IL1. A single polydisperse polypeptide of Mr approximately equal to 80,000 was identified in this way, which bound IL1 alpha and IL1 beta with the same affinity as the IL1-R on intact NOB-1 cells (approximately equal to 10(-10) M). The IL1-binding polypeptide was only seen in membranes from IL1-R-bearing cells and did not react with interleukin 2, tumor necrosis factor alpha, or interferon. IL1-R was purified to apparent homogeneity from solubilized NOB-1 membranes by affinity chromatography on wheat germ agglutinin-Sepharose and IL1 alpha-Sepharose. Gel electrophoresis and silver staining of purified preparations revealed a single protein of Mr approximately equal to 80,000 which reacted positively in the ligand-blotting procedure and which we identify as the ligand-binding moiety of the murine IL1-R. Purified IL1-R exhibited the same affinity and specificity as the receptor on intact cells. The relationship of this protein to proteins identified by covalent cross-linking studies is discussed.

  18. Western blot analyses of measles virus antibody in normal persons and in patients with multiple sclerosis, subacute sclerosing panencephalitis, or atypical measles.

    PubMed Central

    Hankins, R W; Black, F L

    1986-01-01

    A version of the Western blot was developed to detect serum antibodies against measles virus polypeptides. With this technique, a seroepidemiological survey of antibodies to the several measles virus proteins in diverse measles-related conditions was conducted. The sera were obtained from individuals with a recent or long-past history of natural measles, from persons with a history of immunization with live attenuated measles vaccine, and from patients with multiple sclerosis, subacute sclerosing panencephalitis, or atypical measles. The findings indicated that live attenuated measles vaccine elicits an antibody response qualitatively resembling that of a natural infection. In addition, multiple sclerosis patients made less antibody to the measles virus M protein than did individuals with a long-past history of natural measles. Thus, the immunological reaction of multiple sclerosis patients to measles virus is qualitatively, as well as quantitatively, different from that of normal persons. Finally, persons with subacute sclerosing panencephalitis and atypical measles mounted abnormally high antibody responses to measles virus polypeptides, in particular the P protein. PMID:3531224

  19. Determination of the bovine food allergen casein in white wines by quantitative indirect ELISA, SDS-PAGE, Western blot and immunostaining.

    PubMed

    Patrick, Weber; Hans, Steinhart; Angelika, Paschke

    2009-09-23

    This study describes the characterization of allergic bovine casein and caseinate fining agents by SDS-PAGE analysis and the development of a quantitative indirect ELISA for the detection of these substances in wines. The ELISA was applied to various experimental wines that were treated with different caseinate dosages and went through different processing steps and to a panel of commercial wines. Positive results were assured by SDS-PAGE, Western blot, and immunostaining. Comprehensive literature research was done to evaluate the demanded sensitivity of the ELISA. The results showed that alpha- and beta-caseins remain in some wines and are detectable. Estimated amounts were in the range or below an estimated no-observed adverse effect level (NOAEL) of 0.9 mg/L, but it was concluded that there is still an uncertainty about this NOAEL. Additional applied processing, referring to bentonite treatment and successive filtration, was determined to contribute to a significant decrease of casein residues in wines. PMID:19754170

  20. Anti-Peptide Monoclonal Antibodies Generated for Immuno-Multiple Reaction Monitoring-Mass Spectrometry Assays Have a High Probability of Supporting Western blot and ELISA*

    PubMed Central

    Schoenherr, Regine M.; Saul, Richard G.; Whiteaker, Jeffrey R.; Yan, Ping; Whiteley, Gordon R.; Paulovich, Amanda G.

    2015-01-01

    Immunoaffinity enrichment of peptides coupled to targeted, multiple reaction monitoring-mass spectrometry (immuno-MRM) has recently been developed for quantitative analysis of peptide and protein expression. As part of this technology, antibodies are generated to short, linear, tryptic peptides that are well-suited for detection by mass spectrometry. Despite its favorable analytical performance, a major obstacle to widespread adoption of immuno-MRM is a lack of validated affinity reagents because commercial antibody suppliers are reluctant to commit resources to producing anti-peptide antibodies for immuno-MRM while the market is much larger for conventional technologies, especially Western blotting and ELISA. Part of this reluctance has been the concern that affinity reagents generated to short, linear, tryptic peptide sequences may not perform well in traditional assays that detect full-length proteins. In this study, we test the feasibility and success rates of generating immuno-MRM monoclonal antibodies (mAbs) (targeting tryptic peptide antigens) that are also compatible with conventional, protein-based immuno-affinity technologies. We generated 40 novel, peptide immuno-MRM assays and determined that the cross-over success rates for using immuno-MRM monoclonals for Western blotting is 58% and for ELISA is 43%, which compare favorably to cross-over success rates amongst conventional immunoassay technologies. These success rates could most likely be increased if conventional and immuno-MRM antigen design strategies were combined, and we suggest a workflow for such a comprehensive approach. Additionally, the 40 novel immuno-MRM assays underwent fit-for-purpose analytical validation, and all mAbs and assays have been made available as a resource to the community via the Clinical Proteomic Tumor Analysis Consortium's (CPTAC) Antibody (http://antibodies.cancer.gov) and Assay Portals (http://assays.cancer.gov), respectively. This study also represents the first

  1. Earth System Studies in Northern Eurasia

    NASA Astrophysics Data System (ADS)

    Groisman, Pavel; Gulev, Sergey; Maksyutov, Shamil

    2014-04-01

    During recent decades, northern Eurasia (north of 40°N and east of 15°E) was affected by unprecedented climate and environmental changes. Several droughts, including a heat wave in 2010 (that in European Russia may have been the cause of up to an additional 60,000 deaths in July-August of that year, compared to the previous year; see http://ifaran.ru/science/seminars/Summer2010.html); alternated with hazardous extreme precipitation and flood events. Permafrost thaw, retreating Arctic sea ice, increasing areas of forest fire, and dramatic regional warming buffeted this region, tossing northern Eurasia from one extreme condition to the next.

  2. Northern Plains 'Crater'

    NASA Technical Reports Server (NTRS)

    2004-01-01

    10 December 2004 The lower left (southwest) corner of this Mars Global Surveyor (MGS) Mars Orbiter Camera (MOC) image shows the location of a somewhat filled and buried meteor impact crater on the northern plains of Mars. The dark dots are boulders. A portion of a similar feature is seen in the upper right (northeast) corner of the image. This picture, showing landforms (including the odd mound north/northeast of the crater) that are typical of the martian northern lowland plains, was obtained as part of the MGS MOC effort to support the search for a landing site for the Phoenix Mars Scout lander. Phoenix will launch in 2007 and land on the northern plains in 2008. This image is located near 68.0oN, 227.4oW, and covers an area approximately 3 km (1.9 mi) wide. The scene is illuminated by sunlight from the lower left.

  3. Seroprevalence of HIV-type 1 in a northern California health plan population: an unlinked survey.

    PubMed Central

    Hiatt, R A; Capell, F J; Ascher, M S

    1992-01-01

    We conducted an unlinked seroprevalence survey for human immunodeficiency virus-type 1 (HIV-1) of 9821 persons who had a routine personal health appraisal examination in 1989 while members of the Kaiser Permanente Medical Care Program in northern California. An outside laboratory performed enzyme immunoassay (EIA) analyses, and the California Viral and Rickettsial Diseases Laboratory confirmed EIA-reactive samples by immunofluorescent assay and Western blot assay. Only 20 specimens (0.2%) were confirmed as positive, and 18 were from men. These data suggest that, at the time of this survey, HIV-1 infection was not widespread in the northern California population represented by this health plan membership. PMID:1546773

  4. FACING NORTHEAST ACROSS NORTHERN END OF PARK TOWARDS ITS NORTHERN ...

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

    FACING NORTHEAST ACROSS NORTHERN END OF PARK TOWARDS ITS NORTHERN CORNER - Candler Park Historic District, Roughly bounded by Moreland, Dekalb, McLendon & Harold Avenues, Matthews Street & Clifton Terrace, Atlanta, Fulton County, GA

  5. Northern Polar Cap

    NASA Technical Reports Server (NTRS)

    2004-01-01

    [figure removed for brevity, see original site]

    Released 13 May 2004 This nighttime visible color image was collected on November 26, 2002 during the Northern Summer season near the North Polar Cap Edge.

    The THEMIS VIS camera is capable of capturing color images of the martian surface using its five different color filters. In this mode of operation, the spatial resolution and coverage of the image must be reduced to accommodate the additional data volume produced from the use of multiple filters. To make a color image, three of the five filter images (each in grayscale) are selected. Each is contrast enhanced and then converted to a red, green, or blue intensity image. These three images are then combined to produce a full color, single image. Because the THEMIS color filters don't span the full range of colors seen by the human eye, a color THEMIS image does not represent true color. Also, because each single-filter image is contrast enhanced before inclusion in the three-color image, the apparent color variation of the scene is exaggerated. Nevertheless, the color variation that does appear is representative of some change in color, however subtle, in the actual scene. Note that the long edges of THEMIS color images typically contain color artifacts that do not represent surface variation.

    Image information: VIS instrument. Latitude 80, Longitude 43.2 East (316.8 West). 38 meter/pixel resolution.

    Note: this THEMIS visual image has not been radiometrically nor geometrically calibrated for this preliminary release. An empirical correction has been performed to remove instrumental effects. A linear shift has been applied in the cross-track and down-track direction to approximate spacecraft and planetary motion. Fully calibrated and geometrically projected images will be released through the Planetary Data System in accordance with Project policies at a later time.

    NASA's Jet Propulsion Laboratory manages the 2001 Mars Odyssey mission for

  6. New method for simultaneous species-specific identification of equine strongyles (nematoda, strongylida) by reverse line blot hybridization.

    PubMed

    Traversa, Donato; Iorio, Raffaella; Klei, Thomas R; Kharchenko, Vitaliy A; Gawor, Jakub; Otranto, Domenico; Sparagano, Olivier A E

    2007-09-01

    The ability of a reverse line blot (RLB) assay to identify 13 common species of equine small strongyles (cyathostomins) and to discriminate them from three Strongylus spp. (large strongyles) was demonstrated. The assay relied on the specific hybridization of PCR-amplified intergenic spacer DNA fragments of the nuclear ribosomal DNA to membrane-bound species-specific probes. All cyathostomins examined were unequivocally identified and simultaneously discriminated from each other and from three large strongyles (Strongylus edentatus, Strongylus equinus, and Strongylus vulgaris). This assay will enable the accurate and rapid identification of equine cyathostomins irrespective of their life cycle stage, opening important avenues for a better understanding of their biology and epidemiology and of the pathogenesis of cyathostomin-associated disease. In particular, this RLB method promises to be a powerful diagnostic tool to determine the roles of individual species in the pathogenesis of mixed infections and to elucidate some aspects of cyathostominosis. Also, it could represent a basic step toward the development of a rapid and simple molecular test for the early detection of drug-resistant genotypes of horse strongyle species. PMID:17626168

  7. New Method for Simultaneous Species-Specific Identification of Equine Strongyles (Nematoda, Strongylida) by Reverse Line Blot Hybridization▿

    PubMed Central

    Traversa, Donato; Iorio, Raffaella; Klei, Thomas R.; Kharchenko, Vitaliy A.; Gawor, Jakub; Otranto, Domenico; Sparagano, Olivier A. E.

    2007-01-01

    The ability of a reverse line blot (RLB) assay to identify 13 common species of equine small strongyles (cyathostomins) and to discriminate them from three Strongylus spp. (large strongyles) was demonstrated. The assay relied on the specific hybridization of PCR-amplified intergenic spacer DNA fragments of the nuclear ribosomal DNA to membrane-bound species-specific probes. All cyathostomins examined were unequivocally identified and simultaneously discriminated from each other and from three large strongyles (Strongylus edentatus, Strongylus equinus, and Strongylus vulgaris). This assay will enable the accurate and rapid identification of equine cyathostomins irrespective of their life cycle stage, opening important avenues for a better understanding of their biology and epidemiology and of the pathogenesis of cyathostomin-associated disease. In particular, this RLB method promises to be a powerful diagnostic tool to determine the roles of individual species in the pathogenesis of mixed infections and to elucidate some aspects of cyathostominosis. Also, it could represent a basic step toward the development of a rapid and simple molecular test for the early detection of drug-resistant genotypes of horse strongyle species. PMID:17626168

  8. Comparison of trichloroacetic acid with other protein-precipitating agents in enriching abnormal prion protein for Western blot analysis.

    PubMed

    LeBrun, Matthew; Huang, Hongsheng; He, Runtao; Booth, Stephanie; Balachandran, Aru; Li, Xuguang

    2008-06-01

    Detection of the abnormal or the pathogenic form of prion protein (PrP(Sc)) by Western blot (WB) is challenging, especially, for samples derived from cell cultures that contain low levels of PrP(Sc). A variety of PrP(Sc) concentration methods have been reported with various PrP(Sc) recovery efficiencies. Ultracentrifugation is one of the methods used frequently to enrich the pathogenic form of PrP(Sc) prior to WB analyses. The resulting PrP(Sc) pellet is extremely insoluble and often requires sonication to be dissolved, potentially generating aerosols. We modified the common protein-precipitating protocol using trichloroacetic acid to concentrate PrP(Sc) by slow-speed centrifugation, followed by solubilization of the pellets with 6 mol/L urea prior to sodium dodecyl sulphate -- polyacrylamide gel electrophoresis and WB analyses. Comparative studies suggest this simple trichloroacetic acid protocol was more effective in enriching PrP(Sc) presented in cell cultures and brain homogenates than other reported protein-precipitating methods. Furthermore, incorporation of the urea treatment step to dissolve the precipitated PrP(Sc) pellets helped to reduce the infectivity of PrP(Sc). PMID:18535632

  9. Lectin staining and Western blot data showing differential sialylation of nutrient-deprived cancer cells to sialic acid supplementation

    PubMed Central

    Badr, Haitham A.; AlSadek, Dina M.M.; Mathew, Mohit P.; Li, Chen-Zhong; Djansugurova, Leyla B.; Yarema, Kevin J.; Ahmed, Hafiz

    2015-01-01

    This report provides data that are specifically related to the differential sialylation of nutrient deprived breast cancer cells to sialic acid supplementation in support of the research article entitled, “Nutrient-deprived cancer cells preferentially use sialic acid to maintain cell surface glycosylation" [1]. Particularly, breast cancer cells, when supplemented with sialic acid under nutrient deprivation, display sialylated glycans at the cell surface, but non-malignant mammary cells show sialylated glycans intracellularly. The impact of sialic acid supplementation under nutrient deprivation was demonstrated by measuring levels of expression and sialylation of two markers, EGFR1 and MUC1. This Data in Brief article complements the main manuscript by providing detailed instructions and representative results for cell-level imaging and Western blot analyses of changes in sialylation during nutrient deprivation and sialic acid supplementation. These methods can be readily generalized for the study of many types of glycosylation and various glycoprotein markers through the appropriate selection of fluorescently-labeled lectins. PMID:26629491

  10. Simultaneous direct identification of genital microorganisms in voided urine using multiplex PCR-based reverse line blot assays.

    PubMed

    McKechnie, Michelle L; Kong, Fanrong; Gilbert, Gwendolyn L

    2013-01-01

    Our aim was to develop and evaluate sensitive methods that would allow simultaneous direct identification of multiple potential pathogens in clinical specimens for diagnosis and epidemiological studies, using a multiplex PCR-based reverse line blot assay. We have previously developed assays suitable for detection of bacterial respiratory and systemic pathogens. In this chapter we describe, in detail, a method developed to identify 14 genital microorganisms, for use in epidemiological studies of genital infection or colonization, using first voided urine specimens. The 14 urogenital pathogens or putative pathogens studied were Trichomonas vaginalis, Streptococcus pneumoniae, Neisseria gonorrhoeae, N. meningitidis, Chlamydia trachomatis, Ureaplasma parvum, U. urealyticum, Mycoplasma hominis, M. genitalium, Gardnerella vaginalis, Haemophilus influenzae, herpes simplex virus 1 and 2, and adenovirus. Two species-specific primer pairs and probes were designed for each target. The method was validated using a reference strain or a well-characterized clinical isolate of each target organism. In a clinical study among men attending sexual health clinics in Sydney, we used the assay to compare rates of detection of the 14 organisms in men with urethritis with those in asymptomatic controls and found the method to be sensitive, specific, convenient, and relatively inexpensive. PMID:23104293

  11. Immunological Diagnosis of Human Hydatid Cyst Relapse: Utility of the Enzyme-Linked Immunoelectrotransfer Blot and Discriminant Analysis

    PubMed Central

    Gadea, I.; Ayala, G.; Diago, M. T.; Cuñat, A.; Garcia de Lomas, J.

    2000-01-01

    A discriminant technique was applied to the different serological patterns obtained by enzyme-linked immunoelectrotransfer blotting (EITB) and by conventional immunological tests, in order to differentiate the residual antibody patterns present in healed hydatidosis from the ones present in patients with active hydatidosis. For this purpose, specific antibodies against Echinococcus granulosus were detected by indirect hemagglutination, agglutination of latex particles, basophil degranulation, and EITB for 23 patients with active hydatidosis and 45 patients with surgically cured hydatidosis. Discriminant analysis of the different serological patterns obtained by EITB and conventional serology correctly classified 92.54% of patients (93.3% if the patients are differentiated according to the time elapsed since surgery). This method detected the presence of active hydatidosis in 95.6% of patients for whom abdominal ultrasonography had confirmed the presence of active hydatid cysts. The global specificity was 88.9%. The specificity was 97.1% for patients who had been operated on 3 years ago or more and 63.6% for patients with less time since surgery. PMID:10882649

  12. A Modified Quantum Dot-Based Dot Blot Assay for Rapid Detection of Fish Pathogen Vibrio anguillarum.

    PubMed

    Zhang, Yang; Xiao, Jingfan; Wang, Qiyao; Zhang, Yuanxing

    2016-08-28

    Vibrio anguillarum, a devastating pathogen causing vibriosis among marine fish, is prevailing in worldwide fishery industries and accounts for grievous economic losses. Therefore, a rapid on-site detection and diagnostic technique for this pathogen is in urgent need. In this study, two mouse monoclonal antibodies (MAbs) against V. anguillarum, 6B3-C5 and 8G3-B5, were generated by using hybridoma technology and their isotypes were characterized. MAb 6B3-C5 was chosen as the detector antibody and conjugated with quantum dots. Based on MAb 6B3- C5 labeled with quantum dots, a modified dot blot assay was developed for the on-site determination of V. anguillarum. It was found that the method had no cross-reactivity with other than V. anguillarum bacteria. The detection limit (LOD) for V. anguillarum was 1 × 10(3) CFU/ml in cultured bacterial suspension samples, which was a 100-fold higher sensitivity than the reported colloidal gold immunochromatographic test strip. When V. anguillarum was mixed with turbot tissue homogenates, the LOD was 1 × 10(3) CFU/ml, suggesting that tissue homogenates did not influence the detection capabilities. Preenrichment with the tissue homogenates for 12 h could raise the LOD up to 1 × 10(2) CFU/ml, confirming the reliability of the method. PMID:27116991

  13. State-of-the-art housekeeping proteins for quantitative western blotting: Revisiting the first draft of the human proteome.

    PubMed

    Lee, Hyun-Gwan; Jo, Jihoon; Hong, Hyun-Hee; Kim, Kee K; Park, Joong-Ki; Cho, Sung-Jin; Park, Chungoo

    2016-07-01

    Western blotting (WB) analysis is the most popular and widely used methodology for protein detection and characterization over recent decades. In accordance with the advancement of the technologies for the acquisition of WB signals, a quantitative value is used to present the abundance of target proteins in a complex sample, thereby requiring the use of specific proteins as internal references that represent total proteins. Heretofore, proteins encoded by housekeeping genes such as GAPDH, β-tubulin and β-actin have been commonly used as loading controls without any hesitation because their mRNA expression levels tend to be high and constant in many different cells and tissues. Experimentally, however, some of the housekeeping reference proteins are often displayed with inconsistent expression levels in both homogeneous and heterogeneous tissues, and, in terms of mRNA levels, they have a weak correlation to the abundance of proteins. To estimate accurate, reliable, and reproducible protein quantifications, it is crucial to define appropriate reference controls. For this paper, we explored the recently released large-scale, human proteomic database ProteomicsDB including 16 857 liquid chromatography tandem-mass-spectrometry data from 27 human tissues, and suggest 20 ubiquitously- and constitutively-expressed, putative internal-reference controls for the quantification of differential protein expressions. Intriguingly, the most commonly used, known housekeeping genes were entirely excluded in our newly defined candidates. Although the applications of the candidates under many different biological conditions and in other organisms are yet to be empirically verified, we propose reliable, potential loading controls for a WB analysis in this paper. PMID:27125885

  14. Evaluation of a Western Blot and ELISA for the detection of anti-Trichinella-IgG in pig sera.

    PubMed

    Nöckler, K; Reckinger, S; Broglia, A; Mayer-Scholl, A; Bahn, P

    2009-08-26

    Human trichinellosis is a foodborne disease caused by ingestion of infective Trichinella muscle larvae via pork or meat of other food animals which are susceptible to this zoonotic parasite. There are new approaches for a risk-oriented meat inspection for Trichinella in pigs which are accompanied by monitoring programmes on herd level to control freedom from this parasite. For this purpose, testing schemes utilizing serological tests with a high sensitivity and specificity are required. This study aimed at the evaluation of an ELISA and a Western Blot (WB) for the detection of anti-Trichinella-IgG in terms of sensitivity and specificity taking results of artificial digestion as gold standard. For this purpose, 144 field sera from pigs confirmed as Trichinella-free as well as 159 sera from pigs experimentally infected with T. spiralis (123), T. britovi (19) or T. pseudospiralis (17) were examined by ELISA (excretory-secretory antigen) and WB (crude worm extract). Sera from pigs experimentally infected with four other nematode species were included to investigate the cross-reactivity of the antigen used in the WB. For all Trichinella-positive pig sera, band pattern profiles were identified in the WB and results were analysed in relation to ELISA OD% values. Testing of pig sera revealed a sensitivity of 96.8% for the ELISA and 98.1% for the WB whereas the methods showed a specificity of 97.9 and 100%, respectively. WB analysis of Trichinella-positive pig sera revealed five specific band patterns of 43, 47, 61, 66, and 102 kDa of which the 43 kDa protein was identified as the predominant antigen. The frequency of the band pattern profile was irrespective of the dose and the period of infection as well as the Trichinella species investigated. In conclusion, monitoring in swine farms for Trichinella antibodies should be based on screening pig sera by means of ELISA followed by confirmatory testing through WB analysis. PMID:19473770

  15. Molecular mass dependence of hyaluronan detection by sandwich ELISA-like assay and membrane blotting using biotinylated hyaluronan binding protein

    PubMed Central

    Yuan, Han; Tank, Mihir; Alsofyani, Abeer; Shah, Naman; Talati, Nishant; LoBello, Jaclyn C; Kim, Jin Ryoun; Oonuki, Yoji; de la Motte, Carol A; Cowman, Mary K

    2013-01-01

    Hyaluronan (HA) is widely detected in biological samples and its concentration is most commonly determined by the use of a labeled specific HA binding protein (aggrecan G1-IGD-G2, HABP), employing membrane blotting and sandwich enzyme-linked immunosorbent assay (ELISA)-like methods. However, the detected signal intensity or the quantified value obtained by using these surface-based methods is related to the molecular mass (M) of HA, especially for HA in the low M range below ∼150 kDa. At the same mass or mass concentration, higher M HA gives a higher signal than lower M HA. We have experimentally determined the quantitative relationship between the M of HA (in the range 20–150 kDa) and the relative signal intensity in comparison with a standard HA, in a sandwich ELISA-like assay. An M-dependent signal correction factor (SCF) was calculated and used to correct the signal intensity, so that the corrected concentration value would more accurately reflect the true HA concentration in solution. The SCF for polydisperse low M HA was also calculated and compared with experimental results. When the molecular mass distribution of an HA sample is determined by a method such as gel electrophoresis, then its appropriately averaged SCF can be calculated and used to correct the signal in sandwich ELISA to obtain a more accurate concentration estimation. The correction method works for HA with M between ∼150 and 20 kDa, but lower M HA is too poorly detected for useful analysis. The physical basis of the M-dependent detection is proposed to be the increase in detector-accessible fraction of each surface-bound molecule as M increases. PMID:23964097

  16. Reduction of telomeric repeats as a possible predictor for development of hepatocellular carcinoma: convenient evaluation by slot-blot analysis.

    PubMed

    Isokawa, O; Suda, T; Aoyagi, Y; Kawai, H; Yokota, T; Takahashi, T; Tsukada, K; Shimizu, T; Mori, S; Abe, Y; Suzuki, Y; Nomoto, M; Mita, Y; Yanagi, M; Igarashi, H; Asakura, H

    1999-08-01

    Hepatocellular carcinoma (HCC) mainly arises from the liver with chronic inflammation. Because telomere reduction reflects replicative history in somatic cells, we analyzed the possibility that liver tissues surrounding HCC consist of the cells carrying substantial reduction of telomere. We studied 20 HCC and surrounding noncancerous liver tissues (SL) obtained by surgical resection, and 10 laparoscopically obtained needle biopsy specimens of the liver with chronic inflammation including no overt HCC (CI). Five liver tissues without chronic liver diseases (ND) were also examined. Extracted genomic DNAs were blotted on a nylon membrane, and probed at first with radio-labeled d(TTAGGG)(3) and reprobed with radio-labeled d(CCT)(7). The intensity caused by d(TTAGGG)(3) was divided by that of d(CCT)(7). The ratio was defined as telomeric repeats content (TC). Dilution experiments reproducibly revealed almost the same TC. The reduction rate of telomere length through aging estimated by regression analysis of TC was 0.62% per year. Concomitant analyses of TC and average telomere length revealed that both values were significantly correlated (r =.45; P =.009). To compare TC in the liver with respect to chronic inflammation, the value was divided by TC in peripheral blood leukocytes (PBL) from the same donor. The ratio was defined as relative TC (RTC). There was a statistically significant decrease of RTC in CI compared with that in ND (P =.03). Furthermore, RTC in SL was significantly lower than that in CI (P =.0001). These observations suggest that RTC value in liver tissues may digitally indicate a replicative history of hepatocytes under chronic inflammation, and a risk of HCC development. PMID:10421648

  17. Distinctive western blot antibody patterns induced by infection of mice with individual strains of the Mycobacterium avium complex.

    PubMed Central

    Elsaghier, A; Nolan, A; Allen, B; Ivanyi, J

    1992-01-01

    Systemic infection of mice with organisms of the Mycobacterium avium complex (MAC) induced antibody responses, characteristic for each of the three tested individual strains. The influence of host genetic factors was reflected up to 3 months after infection by the finding of generally oligobanded and multibanded Western blot patterns in C57B1/6 and BALB/c mice, respectively. Nevertheless, more bands developed at 6 months in C57BL/6 mice. The response to three antigens of 18,000, 38,000 and 24,000 MW was analysed in greater detail. Antibodies to a protease-resistant 18,000 MW band produced only by BALB/c mice were either strain specific, following infection with M. avium, strain Maa-B2, or cross-reactive within MAC, following infection with M. avium strain Maa-A6 and M. paratuberculosis, strain Map-203. Another protease-resistant antigen of 38,000 MW was immunogenic only in Maa-B2 infected mice. This constituent was found to be related to the protease-sensitive antigen of corresponding molecular weight from M. tuberculosis. Two 24,000 MW proteins of M. paratuberculosis were separated by two-dimensional gel electrophoresis: antibodies to the anodic band were induced by Map-203 infection, whilst the cathodic band was revealed by heteroclitic antibodies from Maa-B2-infected mice. The latter antigen is apparently expressed during in vivo replication, but not during in vitro culture of Maa-B2 bacteria. We generally conclude, that the selective antibody patterns after live infection, could be attributed to differences in the release of native antigens within mycobacterial lesions. In view of a high degree of species specificity, some of the immunogenic constituents identified may also be useful for serodiagnostic application. Images Figure 2 Figure 3 Figure 4 Figure 5 PMID:1526646

  18. Comparison of an indirect fluorescent antibody test with Western blot for the detection of serum antibodies against Encephalitozoon cuniculi in cats.

    PubMed

    Künzel, Frank; Peschke, Roman; Tichy, Alexander; Joachim, Anja

    2014-12-01

    Current clinical research indicates that Encephalitozoon (E.) cuniculi infections in cats may be underdiagnosed, especially in animals with typical ocular signs (cataract/anterior uveitis). Although molecular detection of the pathogen in tissue appears promising, serology remains the major diagnostic tool in the living animal. While serological tests are established for the main host of E. cuniculi, the rabbit, the routine serological diagnosis for cats still needs validation. The aim of the study was to evaluate the consistency of indirect fluorescence antibody test (IFAT) and Western blot (WB) for the detection of IgG antibodies against E. cuniculi in the serum of 84 cats. In addition, PCR of liquefied lens material or intraocular fluid was performed in those of the cats with a suspected ocular E. cuniculi infection. Twenty-one cats with positive PCR results were considered as a positive reference group. Results obtained by IFAT and WB corresponded in 83/84 serum samples, indicating a very good correlation between both serological methods. Using WB as the standard reference, sensitivity and specificity for the detection of antibodies against E. cuniculi by the IFAT were 97.6 and 100%, respectively. The positive and negative predictive values for the IFAT were 100 and 97.7%, respectively. The accuracy (correct classified proportion) for the detection of IgG antibodies against E. cuniculi in cats was 98.8%. The comparison of both serological methods with the PCR results also revealed a good agreement as 20 out of 21 PCR-positive samples were seropositive both in IFAT and WB. Both tests can be considered as equally reliable assays to detect IgG antibodies against E. cuniculi in cats. As the IFAT is quicker and easier to perform, it is recommended for routine use in the diagnosis of feline encephalitozoonosis. PMID:25199557

  19. Northern Meridiani Scene

    NASA Technical Reports Server (NTRS)

    2004-01-01

    19 November 2004 This Mars Global Surveyor (MGS) Mars Orbiter Camera (MOC) image shows eroded remnants of layered sedimentary rock in northern Sinus Meridiani. The layering is best seen in the circular feature at the center/right, which is an old meteor impact crater that was once filled and buried beneath the sedimentary rocks, then later exhumed and eroded to its present state. All of the sedimentary rocks exposed in this portion of northern Sinus Meridiani are probably older than the rocks in central Sinus Meridiani that have been examined this year by the Mars Exploration Rover, Opportunity. Like the rocks visited by the rover, these, too, may contain detailed clues regarding a wetter Mars in the distant past. These landforms are located near 6.0oN, 2.0oW. The image covers an area approximately 3 km (1.9 mi) wide. Sunlight illuminates the scene from the left/lower left.

  20. Development of Double Eastern Blotting for Major Licorice Components, Glycyrrhizin and Liquiritin for Chemical Quality Control of Licorice Using anti-Glycyrrhizin and anti-Liquiritin Monoclonal Antibodies.

    PubMed

    Fujii, Shunsuke; Morinaga, Osamu; Uto, Takuhiro; Nomura, Shuichi; Shoyama, Yukihiro

    2016-02-10

    Licorice is utilized in various food industries around the world for seasoning agents, confectioneries, drinks, and functional foods. Glycyrrhizin (GL) and liquiritin (Liq) are major quality control chemical markers of licorice that have multifunctional bioactivities. Chemical quality control of licorice is important because its component profiles change depending environmental factors (climate, soil condition, and water deficit) and differences between species. Double eastern blotting using anti-GL and anti-Liq monoclonal antibodies was developed for more convenient, rapid, and specific quality control analysis of GL and Liq, respectively. Moreover, double eastern blotting was applied to investigate the immunohistochemical distributions of GL and Liq in the root of fresh licorice; the localization of both components was then clarified visually. This double eastern blotting technique for GL and Liq may serve as a powerful approach for visually determining the chemical quality of licorice. PMID:26765784

  1. Northern Plains Patterns

    NASA Technical Reports Server (NTRS)

    2003-01-01

    MGS MOC Release No. MOC2-513, 14 October 2003

    Patterns are common on the northern plains of Mars. Like their terrestrial counterparts in places like Siberia, Alaska, and northern Canada, patterned ground on Mars might be an indicator of the presence of ground ice. Whether it is true that the patterns on Mars are related to ground ice and whether the ice is still present beneath the martian surface are unknown. This Mars Global Surveyor (MGS) Mars Orbiter Camera (MOC) picture shows an example of patterned ground on the martian northern plains near 72.4oN, 252.6oW. The dark dots and lines are low mounds and chains of mounds. The circular feature near the center of the image is the location of a buried meteor impact crater; its presence today is marked only by the dark boulders on its rim and ejecta blanket that have managed to remain uncovered at the martian surface. The area shown is 3 km (1.9 mi) wide and illuminated by sunlight from the lower left.

  2. Identification of recombinant human EPO variants in greyhound plasma and urine by ELISA, LC-MS/MS and western blotting: a comparative study.

    PubMed

    Timms, Mark; Steel, Rohan; Vine, John

    2016-02-01

    The recombinant human erythropoietins epoetin alfa (Eprex®), darbepoetin (Aranesp®) and methoxy polyethylene glycol-epoetin beta (Mircera®) were administered to greyhounds for 7, 10 and 14 days respectively. Blood and urine samples were collected and analysed for erythropoietin by ELISA, LC-MS/MS and western blotting. Limits of confirmation in plasma for western blotting and LC-MS/MS methods ranged from a low of 2.5mIU/mL, and closely matched the sensitivity of ELISA screening. PMID:26290355

  3. Characterization of biotin-labeled proteoglycans by electrophoretic separation on minigels and blotting onto nylon membranes prior and after enzymatic digestion.

    PubMed

    Stöcker, G; Lückge, J; Greiling, H; Wagener, C

    1989-06-01

    Biotinylated proteoglycans were separated by sodium dodecyl sulfate electrophoresis prior and after enzymatic digestion by glycan-specific enzymes using polyacrylamide minigels. The biotin-labeled compounds were blotted onto nylon membranes either by electrophoresis or by diffusion and detected by avidin-enzyme conjugates. The method allows the nonisotopic detection of native proteoglycans and core proteins. Proteoglycans can be visualized at protein amounts as low as 0.7 ng per lane. In comparison with sensitive protein stains, compounds of enzyme preparations do not interfere with bands corresponding to core proteins. Electrophoresis, blotting, and staining of up to 12 samples per gel are accomplished in less than 3 h. PMID:2505636

  4. Evaluation of two sets of immunohistochemical and Western blot confirmatory methods in the detection of typical and atypical BSE cases

    PubMed Central

    2011-01-01

    Background Three distinct forms of bovine spongiform encephalopathy (BSE), defined as classical (C-), low (L-) or high (H-) type, have been detected through ongoing active and passive surveillance systems for the disease. The aim of the present study was to compare the ability of two sets of immunohistochemical (IHC) and Western blot (WB) BSE confirmatory protocols to detect C- and atypical (L- and H-type) BSE forms. Obex samples from cases of United States and Italian C-type BSE, a U.S. H-type and an Italian L-type BSE case were tested in parallel using the two IHC sets and WB methods. Results The two IHC techniques proved equivalent in identifying and differentiating between C-type, L-type and H-type BSE. The IHC protocols appeared consistent in the identification of PrPSc distribution and deposition patterns in relation to the BSE type examined. Both IHC methods evidenced three distinct PrPSc phenotypes for each type of BSE: prevailing granular and linear tracts pattern in the C-type; intraglial and intraneuronal deposits in the H-type; plaques in the L-type. Also, the two techniques gave comparable results for PrPSc staining intensity on the C- and L-type BSE samples, whereas a higher amount of intraglial and intraneuronal PrPSc deposition on the H-type BSE case was revealed by the method based on a stronger demasking step. Both WB methods were consistent in identifying classical and atypical BSE forms and in differentiating the specific PrPSc molecular weight and glycoform ratios of each form. Conclusions The study showed that the IHC and WB BSE confirmatory methods were equally able to recognize C-, L- and H-type BSE forms and to discriminate between their different immunohistochemical and molecular phenotypes. Of note is that for the first time one of the two sets of BSE confirmatory protocols proved effective in identifying the L-type BSE form. This finding helps to validate the suitability of the BSE confirmatory tests for BSE surveillance currently in

  5. Femtogram detection of cytokines in a direct dot-blot assay using SERS microspectroscopy and hydrophilically stabilized Au-Ag nanoshells.

    PubMed

    Wang, Yuling; Salehi, Mohammad; Schütz, Max; Schlücker, Sebastian

    2014-03-14

    Rapid parallel detection of two cytokines (IL-6 and IL-8) with femtogram sensitivity in a simple direct dot-blot assay is demonstrated. The microspectroscopic SERS acquisition scheme employs rationally designed, hydrophilically stabilized Au-Ag nanoshells as SERS labels, which are optimized for signal enhancement upon red laser excitation. PMID:24398564

  6. Resolution and identification of major peanut allergens using a combination of fluorescence two-dimensional differential gel electrophoresis, western blotting and Q-TOF mass spectrometry.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Peanut allergy is triggered by several proteins known as allergens. The matching resolution and identification of major peanut allergens in 2D protein maps, was accomplished by the use of fluorescence two-dimensional differential gel electrophoresis (2D DIGE), Western blotting and quadrupole time-of...

  7. Profile of the MP Diagnostics HTLV Blot 2.4 test: a supplemental assay for the confirmation and differentiation of antibodies to HTLV-1 and HTLV-2.

    PubMed

    Miller, Liane

    2016-01-01

    As the first US FDA-approved assay for supplemental HTLV testing, the MP Diagnostics HTLV Blot 2.4 is an effective and efficient method for confirming and differentiating HTLV type infection in repeatedly reactive samples. Novel and patented antigens added increased sensitivity in identifying specimens from infected individuals while differentiating those from uninfected individuals with false reactivity. PMID:26589659

  8. Fires in Northern Australia

    NASA Technical Reports Server (NTRS)

    2002-01-01

    Several fires were detected in Northern Australia by MODIS. The fires show up as red dots, superimposed on a surface reflectance product. The image also shows the Clarence Strait, which separates the mainland from Melville Island to the northwest and the smaller Bathurst Island to its west. The Strait connects the more confined, bowl-shaped Van Diemen Gulf to the Beagle Gulf. To the right of the image at the top is the Gulf of Carpentaria, which appears to be full of phytoplankton, as evidenced by the blue-green swirls in the waters

  9. Northern plants and ozone.

    PubMed

    Manninen, Sirkku; Huttunen, Satu; Tømmervik, Hans; Hole, Lars R; Solberg, Sverre

    2009-12-01

    Forests in northern Fennoscandia are mainly composed of the O3-sensitive species--Scots pine and downy, mountain, and silver birches. Seminatural vegetation also contributes to biodiversity, carbon cycling, and ecosystem services as a part of forests, mires, meadows, and road verges. Fumigation experiments show that current O3 concentrations of 30-50 ppb reduce plant biomass production and reproduction. Visible foliar injury is attributable to peak O3 concentrations and relates to fast phenological development and high growth rate. Trees can acclimate to O3-induced water stress by producing more xeromorphic leaves or needles. The direct effects of O3 on grassland vegetation also translate to changes in the structure and size of the soil microbial community, and ecosystem N cycling. It is necessary to reduce the emission of O3 precursors and maintain high biodiversity to protect northern ecosystems. Regular, systematic, countrywide monitoring and validation as well as quantification of the effects of O3 on plants in the Nordic countries are also necessary. PMID:20175438

  10. Comparative reproductive and physiological responses of northern bobwhite and scaled quail to water deprivation

    USGS Publications Warehouse

    Giuliano, W.M.; Patino, R.; Lutz, R.S.

    1998-01-01

    We compared reproductive and physiological responses of captive female northern bobwhite (Colinus virginianus) and scaled quail (Callipepla squamata) under control and water deprivation conditions. Scaled quail required less food and water to reproduce successfully under control conditions than northern bobwhite. Additionally, in scaled quail, serum osmolality levels and kidney mass were unaffected by water deprivation, whereas in northern bobwhite, serum osmolality levels increased and kidney mass declined. This finding indicates that scaled quail may have osmoregulatory abilities superior to those of northern bobwhite. Under control conditions, northern bobwhite gained more body mass and produced more but smaller eggs than scaled quail. Under water deprivation conditions, northern bobwhite lost more body mass but had more laying bens with a higher rate of egg production than scaled quail. Our data suggest that northern bobwhite allocated more resources to reproduction than to body maintenance, while scaled quail apparently forego reproduction in favor of body maintenance during water deprivation conditions.

  11. Smoke Blankets Northern California

    NASA Technical Reports Server (NTRS)

    2008-01-01

    Lightning strikes have sparked more than a thousand fires in northern California. This image was captured by the Multi-angle Imaging SpectroRadiometer's nadir (vertical-viewing) camera on 27 June 2008. Cape Mendocino is at the center of the image and Mt. Shasta is near the upper right. Concentrated smoke is visible in several river valleys and the large smoke cloud extends over the Pacific Ocean for hundreds of kilometers.

    MISR was built and is managed by NASA's Jet Propulsion Laboratory, Pasadena, Calif., for NASA's Science Mission Directorate, Washington, D.C. The Terra satellite is managed by NASA's Goddard Space Flight Center, Greenbelt, Md. JPL is a division of the California Institute of Technology.

  12. Northern Sinus Meridiani Stereo

    NASA Technical Reports Server (NTRS)

    2003-01-01

    MGS MOC Release No. MOC2-341, 25 April 2003

    This is a stereo (3-d anaglyph) composite of Mars Global Surveyor (MGS) Mars Orbiter Camera (MOC) wide angle images of northern Sinus Meridiani near 2oN, 0oW. The light-toned materials at the south (bottom) end of the picture are considered to be thick (100-200 meters; 300-600 ft) exposures of sedimentary rock. Several ancient meteor impact craters are being exhumed from within these layered materials. To view in stereo, use '3-d' glasses with red over the left eye, and blue over the right. The picture covers an area approximately 113 km (70 mi) wide; north is up.

  13. Teleradiology in northern Quebec.

    PubMed

    Pagé, G; Grégoire, A; Galand, C; Sylvestre, J; Chahlaoui, J; Fauteux, P; Dussault, R; Séguin, R; Roberge, F A

    1981-08-01

    A two-way television network using the Canadian satellite ANIK-B was utilized to transmit radiographic images from Northern Quebec to Montreal. The accuracy of the radiologist's interpretation and his satisfaction with the TV system were studied using a series of 67 preselected cases and 425 current clinical cases. The four participating radiologists gave correct TV interpretations in 81% of the 39 selected cases presented at the beginning of the experiment. This value reached 94% for the other 28 selected cases presented after three months of regular use of the TV system. With current clinical cases, the agreement between TV and direct interpretations was 93%. Although magnification was available, correct identification of very small lesions proved to be the major source of error. On the whole, the radiologists were satisfied with the TV system. PMID:7255709

  14. Performance of a Western blot assay to compare mother and newborn anti-Toxoplasma antibodies for the early neonatal diagnosis of congenital toxoplasmosis.

    PubMed

    Robert-Gangneux, F; Commerce, V; Tourte-Schaefer, C; Dupouy-Camet, J

    1999-09-01

    The aim of this study was to retrospectively evaluate the performance of a Western blot assay to compare mother and newborn anti-Toxoplasma gondii antibodies for the early neonatal diagnosis of congenital toxoplasmosis. Since specific anti-Toxoplasma IgM or IgA is detected inconstantly at birth in the neonate, the diagnosis of congenital toxoplasmosis is often delayed until 6-9 months, after IgG titers have been observed persistently. In this study, 81 paired samples from 60 mother/child pairs were tested for IgG and IgM patterns. All mothers had (or were strongly suspected to have) acquired toxoplasmosis during pregnancy. Specific IgM and IgA were simultaneously detected by immunocapture tests, and IgG was titrated. A serological and clinical follow-up of infants was conducted during the first year of life until the diagnosis of congenital toxoplasmosis could be either confirmed or ruled out. Seventeen of the 60 newborns were congenitally infected. Specific IgM or IgA was detected by immunocapture at birth in 76.5% and 70.6% of cord sera from infected neonates, respectively, with an equal specificity of 77.5%. Comparative Western blot allowed the detection of neosynthesized IgG and IgM in the cord blood of 50% and 78.6% of infected infants, respectively, with a specificity of 100%. The combination of IgA and IgM immunocapture tests, the analysis of IgG and IgM Western blot patterns, and the combination of both techniques allowed the detection of 94%, 94%, and 100% of cases within the first 3 months of life, respectively. In conclusion, Western blotting seems to be a useful complementary tool for the early postnatal diagnosis of congenital toxoplasmosis. PMID:10534187

  15. Rapid fluorescent monitoring of total protein patterns on sodium dodecyl sulfate-polyacrylamide gels and western blots before immunodetection and sequencing.

    PubMed

    Alba, F J; Daban, J R

    1998-10-01

    The fluorogenic dye 2-methoxy-2,4-diphenyl-3(2H)-furanone (MDPF) has been used for the detection of total protein patterns on polyvinylidene difluoride (PVDF) membranes. Fluorescent staining of protein bands on membranes with this covalent dye is completed in 20 min. Wet membranes are translucent, allowing protein visualization by transillumination with ultraviolet light. The resulting images can be recorded using Polaroid film or a charge-coupled device camera. Electrophoretic bands containing 5-10 ng of protein can be detected on the MDPF-stained Western blot. When proteins are directly transferred to the membrane using a slot blotting device, as little as 0.5 ng of protein can be detected. Previous visualization of protein bands on sodium dodecyl sulfate-polyacrylamide gels with the noncovalent fluorescent dye Nile red (Alba et al., BioTechniques, 1996, 21, 625-626) does not interfere with further MDPF staining and fluorescent detection of these bands transferred to PVDF membranes. Thus, Nile red and MDPF staining can be performed sequentially, allowing the rapid monitoring of total protein patterns on both the electrophoretic gel and Western blot. Using the conditions described in this study, MDPF staining does not preclude further N-terminal microsequencing and immunodetection of specific bands with polyclonal antibodies. PMID:9820958

  16. Anti-RAINBOW dye-specific antibodies as universal tools for the visualization of prestained protein molecular weight markers in Western blot analysis

    PubMed Central

    Schüchner, Stefan; Andorfer, Peter; Mudrak, Ingrid; Ogris, Egon

    2016-01-01

    Western blotting is one of the most widely used techniques in molecular biology and biochemistry. Prestained proteins are used as molecular weight standards in protein electrophoresis. In the chemiluminescent Western blot analysis, however, these colored protein markers are invisible leaving researchers with the unsatisfying situation that the signal for the protein of interest and the signal for the markers are not captured simultaneously and have to be merged in an error-prone step. To allow the simultaneous detection of marker proteins we generated monoclonal antibodies specific for the protein dyes. To elicit a dye rather than protein specific immune response we immunized mice sequentially with dye-carrier protein complexes, in which a new carrier protein was used for each subsequent immunization. Moreover, by sequentially immunizing with dye-carrier protein complexes, in which different but structurally related dyes were used, we could also generate an antibody, termed anti-RAINBOW, that cross-reacted even with structurally related dyes not used in the immunizations. Our novel antibodies represent convenient tools for the simultaneous Western blot detection of commercially available prestained marker proteins in combination with the detection of any specific protein of interest. These antibodies will render obsolete the anachronistic tradition of manually charting marker bands on film. PMID:27531616

  17. Anti-RAINBOW dye-specific antibodies as universal tools for the visualization of prestained protein molecular weight markers in Western blot analysis.

    PubMed

    Schüchner, Stefan; Andorfer, Peter; Mudrak, Ingrid; Ogris, Egon

    2016-01-01

    Western blotting is one of the most widely used techniques in molecular biology and biochemistry. Prestained proteins are used as molecular weight standards in protein electrophoresis. In the chemiluminescent Western blot analysis, however, these colored protein markers are invisible leaving researchers with the unsatisfying situation that the signal for the protein of interest and the signal for the markers are not captured simultaneously and have to be merged in an error-prone step. To allow the simultaneous detection of marker proteins we generated monoclonal antibodies specific for the protein dyes. To elicit a dye rather than protein specific immune response we immunized mice sequentially with dye-carrier protein complexes, in which a new carrier protein was used for each subsequent immunization. Moreover, by sequentially immunizing with dye-carrier protein complexes, in which different but structurally related dyes were used, we could also generate an antibody, termed anti-RAINBOW, that cross-reacted even with structurally related dyes not used in the immunizations. Our novel antibodies represent convenient tools for the simultaneous Western blot detection of commercially available prestained marker proteins in combination with the detection of any specific protein of interest. These antibodies will render obsolete the anachronistic tradition of manually charting marker bands on film. PMID:27531616

  18. [Food additives and healthiness].

    PubMed

    Heinonen, Marina

    2014-01-01

    Additives are used for improving food structure or preventing its spoilage, for example. Many substances used as additives are also naturally present in food. The safety of additives is evaluated according to commonly agreed principles. If high concentrations of an additive cause adverse health effects for humans, a limit of acceptable daily intake (ADI) is set for it. An additive is a risk only when ADI is exceeded. The healthiness of food is measured on the basis of nutrient density and scientifically proven effects. PMID:24772784

  19. Polyimide processing additives

    NASA Technical Reports Server (NTRS)

    Pratt, J. R.; St. Clair, T. L.; Burks, H. D.; Stoakley, D. M.

    1987-01-01

    A method has been found for enhancing the melt flow of thermoplastic polyimides during processing. A high molecular weight 422 copoly(amic acid) or copolyimide was fused with approximately 0.05 to 5 pct by weight of a low molecular weight amic acid or imide additive, and this melt was studied by capillary rheometry. Excellent flow and improved composite properties on graphite resulted from the addition of a PMDA-aniline additive to LARC-TPI. Solution viscosity studies imply that amic acid additives temporarily lower molecular weight and, hence, enlarge the processing window. Thus, compositions containing the additive have a lower melt viscosity for a longer time than those unmodified.

  20. Telemedicine in northern Quebec.

    PubMed Central

    Roberge, F. A.; Pagé, G.; Sylvestre, J.; Chahlaoui, J.

    1982-01-01

    Television transmission of diagnostic and educational information can help to improve specialized medical care in remote and underserviced areas. This paper describes a pilot study in which the Canadian satellite Anik-B was used to link the James Bay area in northern Quebec with two large Montreal teaching hospitals. Broad-band real-time television was well suited for tele-education and teleconsultation activities. A much less costly method, using narrow-band slow-scan television, was also examined, but it requires improvements. The technology of telemedicine is in place, but its future use is impeded by the prohibitive costs of operating an efficient two-way broad-band television system for several remote health care sites. A solution to this problem may be an intermediate-band system combining some of the low-cost features of narrowband slow-scan television with the interactive high-resolution advantages of broad-band real-time television. PMID:7139483

  1. Additive usage levels.

    PubMed

    Langlais, R

    1996-01-01

    With the adoption of the European Parliament and Council Directives on sweeteners, colours and miscellaneous additives the Commission is now embarking on the project of coordinating the activities of the European Union Member States in the collection of the data that are to make up the report on food additive intake requested by the European Parliament. This presentation looks at the inventory of available sources on additive use levels and concludes that for the time being national legislation is still the best source of information considering that the directives have yet to be transposed into national legislation. Furthermore, this presentation covers the correlation of the food categories as found in the additives directives with those used by national consumption surveys and finds that in a number of instances this correlation still leaves a lot to be desired. The intake of additives via food ingestion and the intake of substances which are chemically identical to additives but which occur naturally in fruits and vegetables is found in a number of cases to be higher than the intake of additives added during the manufacture of foodstuffs. While the difficulties are recognized in contributing to the compilation of food additive intake data, industry as a whole, i.e. the food manufacturing and food additive manufacturing industries, are confident that in a concerted effort, use data on food additives by industry can be made available. Lastly, the paper points out that with the transportation of the additives directives into national legislation and the time by which the food industry will be able to make use of the new food legislative environment several years will still go by; food additives use data by the food industry will thus have to be reviewed at the beginning of the next century. PMID:8792135

  2. An additional middle cuneiform?

    PubMed Central

    Brookes-Fazakerley, S.D.; Jackson, G.E.; Platt, S.R.

    2015-01-01

    Additional cuneiform bones of the foot have been described in reference to the medial bipartite cuneiform or as small accessory ossicles. An additional middle cuneiform has not been previously documented. We present the case of a patient with an additional ossicle that has the appearance and location of an additional middle cuneiform. Recognizing such an anatomical anomaly is essential for ruling out second metatarsal base or middle cuneiform fractures and for the preoperative planning of arthrodesis or open reduction and internal fixation procedures in this anatomical location. PMID:26224890

  3. Carbamate deposit control additives

    SciTech Connect

    Honnen, L.R.; Lewis, R.A.

    1980-11-25

    Deposit control additives for internal combustion engines are provided which maintain cleanliness of intake systems without contributing to combustion chamber deposits. The additives are poly(oxyalkylene) carbamates comprising a hydrocarbyloxyterminated poly(Oxyalkylene) chain of 2-5 carbon oxyalkylene units bonded through an oxycarbonyl group to a nitrogen atom of ethylenediamine.

  4. MISR Views Northern Australia

    NASA Technical Reports Server (NTRS)

    2000-01-01

    MISR images of tropical northern Australia acquired on June 1, 2000 (Terra orbit 2413) during the long dry season. Left: color composite of vertical (nadir) camera blue, green, and red band data. Right: multi-angle composite of red band data only from the cameras viewing 60 degrees aft, 60 degrees forward, and nadir. Color and contrast have been enhanced to accentuate subtle details. In the left image, color variations indicate how different parts of the scene reflect light differently at blue, green, and red wavelengths; in the right image color variations show how these same scene elements reflect light differently at different angles of view. Water appears in blue shades in the right image, for example, because glitter makes the water look brighter at the aft camera's view angle. The prominent inland water body is Lake Argyle, the largest human-made lake in Australia, which supplies water for the Ord River Irrigation Area and the town of Kununurra (pop. 6500) just to the north. At the top is the southern edge of Joseph Bonaparte Gulf; the major inlet at the left is Cambridge Gulf, the location of the town of Wyndham (pop. 850), the port for this region. This area is sparsely populated, and is known for its remote, spectacular mountains and gorges. Visible along much of the coastline are intertidal mudflats of mangroves and low shrubs; to the south the terrain is covered by open woodland merging into open grassland in the lower half of the pictures.

    MISR was built and is managed by NASA's Jet Propulsion Laboratory, Pasadena, CA, for NASA's Office of Earth Science, Washington, DC. The Terra satellite is managed by NASA's Goddard Space Flight Center, Greenbelt, MD. JPL is a division of the California Institute of Technology.

  5. Isolated Northern Dunes

    NASA Technical Reports Server (NTRS)

    2005-01-01

    [figure removed for brevity, see original site]

    Our topic for the weeks of April 4 and April 11 is dunes on Mars. We will look at the north polar sand sea and at isolated dune fields at lower latitudes. Sand seas on Earth are often called 'ergs,' an Arabic name for dune field. A sand sea differs from a dune field in two ways: 1) a sand sea has a large regional extent, and 2) the individual dunes are large in size and complex in form.

    This VIS image was taken at 81 degrees North latitude during Northern spring. In this region, the dunes are isolated from each other. The dunes are just starting to emerge from the winter frost covering appearing dark with bright crests. These dunes are located on top of ice.

    Image information: VIS instrument. Latitude 82.1, Longitude 191.3 East (168.7 West). 19 meter/pixel resolution.

    Note: this THEMIS visual image has not been radiometrically nor geometrically calibrated for this preliminary release. An empirical correction has been performed to remove instrumental effects. A linear shift has been applied in the cross-track and down-track direction to approximate spacecraft and planetary motion. Fully calibrated and geometrically projected images will be released through the Planetary Data System in accordance with Project policies at a later time.

    NASA's Jet Propulsion Laboratory manages the 2001 Mars Odyssey mission for NASA's Office of Space Science, Washington, D.C. The Thermal Emission Imaging System (THEMIS) was developed by Arizona State University, Tempe, in collaboration with Raytheon Santa Barbara Remote Sensing. The THEMIS investigation is led by Dr. Philip Christensen at Arizona State University. Lockheed Martin Astronautics, Denver, is the prime contractor for the Odyssey project, and developed and built the orbiter. Mission operations are conducted jointly from Lockheed Martin and from JPL, a division of the California Institute of Technology in Pasadena.

  6. Northern Sand Sea

    NASA Technical Reports Server (NTRS)

    2005-01-01

    [figure removed for brevity, see original site]

    Our topic for the weeks of April 4 and April 11 is dunes on Mars. We will look at the north polar sand sea and at isolated dune fields at lower latitudes. Sand seas on Earth are often called 'ergs,' an Arabic name for dune field. A sand sea differs from a dune field in two ways: 1) a sand sea has a large regional extent, and 2) the individual dunes are large in size and complex in form.

    This VIS image was taken at 82 degrees North latitude during Northern spring. The image is completely dominated by dunes. In sand seas, it is very common for a single type of dune to occur, and for a single predominate wind to control the alignment of the dunes.

    Image information: VIS instrument. Latitude 82.2, Longitude 152.5 East (207.5 West). 19 meter/pixel resolution.

    Note: this THEMIS visual image has not been radiometrically nor geometrically calibrated for this preliminary release. An empirical correction has been performed to remove instrumental effects. A linear shift has been applied in the cross-track and down-track direction to approximate spacecraft and planetary motion. Fully calibrated and geometrically projected images will be released through the Planetary Data System in accordance with Project policies at a later time.

    NASA's Jet Propulsion Laboratory manages the 2001 Mars Odyssey mission for NASA's Office of Space Science, Washington, D.C. The Thermal Emission Imaging System (THEMIS) was developed by Arizona State University, Tempe, in collaboration with Raytheon Santa Barbara Remote Sensing. The THEMIS investigation is led by Dr. Philip Christensen at Arizona State University. Lockheed Martin Astronautics, Denver, is the prime contractor for the Odyssey project, and developed and built the orbiter. Mission operations are conducted jointly from Lockheed Martin and from JPL, a division of the California Institute of Technology in Pasadena.

  7. Reverse Cross Blot Hybridization Assay for Rapid Detection of PCR-Amplified DNA from Candida Species, Cryptococcus neoformans, and Saccharomyces cerevisiae in Clinical Samples

    PubMed Central

    Posteraro, Brunella; Sanguinetti, Maurizio; Masucci, Luca; Romano, Lucio; Morace, Giulia; Fadda, Giovanni

    2000-01-01

    A PCR-based assay was developed to detect and identify medically important yeasts in clinical samples. Using a previously described set of primers (G. Morace et al., J. Clin. Microbiol. 35:667–672, 1997), we amplified a fragment of the ERG11 gene for cytochrome P-450 lanosterol 14α-demethylase, a crucial enzyme in the biosynthesis of ergosterol. The PCR product was analyzed in a reverse cross blot hybridization assay with species-specific probes directed to a target region of the ERG11 gene of Candida albicans (pCal), C. guilliermondii (pGui), C. (Torulopsis) glabrata (pGla), C. kefyr (pKef), C. krusei (pKru), C. parapsilosis (pPar), C. tropicalis (pTro), the newly described species C. dubliniensis (pDub), Saccharomyces cerevisiae (pSce), and Cryptococcus neoformans (pCry). The PCR-reverse cross blot hybridization assay correctly identified multiple isolates of each species tested. No cross-hybridization was detected with any other fungal, bacteria, or human DNAs tested. The method was tested against conventional identification on 140 different clinical samples, including blood and cerebrospinal fluid, from patients with suspected fungal infections. The results agreed with those of culture and phenotyping for all but six specimens (two of which grew yeasts not included in the PCR panel of probes and four in which PCR positivity-culture negativity was justified by clinical findings). Species identification time was reduced from a mean of 4 days with conventional identification to 7 h with the molecular method. The PCR-reverse cross blot hybridization assay is a rapid method for the direct detection and identification of yeasts in clinical samples. PMID:10747151

  8. The epidemiology of tick-borne haemoparasites as determined by the reverse line blot hybridization assay in an intensively studied cohort of calves in western Kenya

    PubMed Central

    Njiiri, Nyawira E.; Bronsvoort, B. Mark deC.; Collins, Nicola E.; Steyn, Helena C.; Troskie, Milana; Vorster, Ilse; Thumbi, S.M.; Sibeko, Kgomotso P.; Jennings, Amy; van Wyk, Ilana Conradie; Mbole-Kariuki, Mary; Kiara, Henry; Poole, E. Jane; Hanotte, Olivier; Coetzer, Koos; Oosthuizen, Marinda C.; Woolhouse, Mark; Toye, Philip

    2015-01-01

    The development of sensitive surveillance technologies using PCR-based detection of microbial DNA, such as the reverse line blot assay, can facilitate the gathering of epidemiological information on tick-borne diseases, which continue to hamper the productivity of livestock in many parts of Africa and elsewhere. We have employed a reverse line blot assay to detect the prevalence of tick-borne parasites in an intensively studied cohort of indigenous calves in western Kenya. The calves were recruited close to birth and monitored for the presence of infectious disease for up to 51 weeks. The final visit samples from 453 calves which survived for the study period were analyzed by RLB. The results indicated high prevalences of Theileria mutans (71.6%), T. velifera (62.8%), Anaplasma sp. Omatjenne (42.7%), A. bovis (39.9%), Theileria sp. (sable) (32.7%), T. parva (12.9%) and T. taurotragi (8.5%), with minor occurrences of eight other haemoparasites. The unexpectedly low prevalence of the pathogenic species Ehrlichia ruminantium was confirmed by a species-specific PCR targeting the pCS20 gene region. Coinfection analyses of the seven most prevalent haemoparasites indicated that they were present as coinfections in over 90% of the cases. The analyses revealed significant associations between several of the Theileria parasites, in particular T. velifera with Theileria sp. sable and T. mutans, and T. parva with T. taurotragi. There was very little coinfection of the two most common Anaplasma species, although they were commonly detected as coinfections with the Theileria parasites. The comparison of reverse line blot and serological results for four haemoparasites (T. parva, T. mutans, A. marginale and B. bigemina) indicated that, except for the mostly benign T. mutans, indigenous cattle seem capable of clearing infections of the three other, pathogenic parasites to below detectable levels. Although the study site was located across four agroecological zones, there was

  9. Comparing the Southern blot method and polymerase chain reaction product analysis for chimeric RCCX detection in CYP21A2 deficiency.

    PubMed

    Lee, Hsien-Hsiung; Lee, Yann-Jinn; Chao, Mei-Chyn

    2010-04-15

    The 3.2-kb TaqI-produced fragment of the CYP21A1P pseudogene and the 3.7-kb TaqI-produced fragment of the functional CYP21A2 gene exist on chromosome 6p21.3. We used the polymerase chain reaction (PCR) product and Southern blot method with TaqI endonuclease digestion to identify a chimeric RCCX module in two unrelated patients with congenital adrenal hyperplasia (CAH). After TaqI cleavage, the PCR product analysis revealed that patient 1 with the chimeric CYP21A1P/CYP21A2 gene in one allele and IVS2-12A/C>G in combination with the 707-714del mutation in the other allele produced a configuration of 3.2- and 2.4-kb fragments. Patient 2, who carried IVS2-12A/C>G in combination with the 707-714del mutation in one allele and the chimeric TNXA/TNXB gene in the other allele, presented with 3.2- and 2.3-kb fragments. However, Southern blot analysis showed that patients 1 and 2 produced 3.2-, 2.4-, and 2.5-kb fragments. We conclude that the chimeric CYP21A1P/CYP21A2 gene, IVS2-12A/C>G in combination with the 707-714del mutation, and the chimeric TNXA/TNXB gene cannot be distinguished by the Southern blot method. Conversely, the chimeric TNXA/TNXB gene was identified in the PCR product analysis due to the appearance of the 2.37-kb fragment, which indicates the occurrence of the chimeric TNXA/TNXB formation extending to the boundary of TNXA in the RCCX region. PMID:19961824

  10. Use of the water-soluble fluor sodium salicylate for fluorographic detection of tritium in thin-layer chromatograms and nitrocellulose blots

    SciTech Connect

    Lucher, L.A.; Lego, T.

    1989-05-01

    We have determined that sodium salicylate, a water-soluble fluor which we use routinely for fluorography with polyacrylamide gels, is also useful for fluorography with thin-layer media. Detection of /sup 3/H-labeled material applied to thin-layer chromatography plates, or nitrocellulose membranes, can be enhanced up to 150-fold after treatment with an aqueous solution of 2 M sodium salicylate, while detection of /sup 35/S-labeled material is enhanced only about 2-fold. We demonstrate the utility of sodium salicylate fluorography in detecting 3H-labeled palmitic acid following thin-layer chromatography and /sup 3/H-labeled proteins following blotting to nitrocellulose.

  11. Polyimide processing additives

    NASA Technical Reports Server (NTRS)

    Fletcher, James C. (Inventor); Pratt, J. Richard (Inventor); St.clair, Terry L. (Inventor); Stoakley, Diane M. (Inventor); Burks, Harold D. (Inventor)

    1992-01-01

    A process for preparing polyimides having enhanced melt flow properties is described. The process consists of heating a mixture of a high molecular weight poly-(amic acid) or polyimide with a low molecular weight amic acid or imide additive in the range of 0.05 to 15 percent by weight of additive. The polyimide powders so obtained show improved processability, as evidenced by lower melt viscosity by capillary rheometry. Likewise, films prepared from mixtures of polymers with additives show improved processability with earlier onset of stretching by TMA.

  12. Polyimide processing additives

    NASA Technical Reports Server (NTRS)

    Pratt, J. Richard (Inventor); St.clair, Terry L. (Inventor); Stoakley, Diane M. (Inventor); Burks, Harold D. (Inventor)

    1993-01-01

    A process for preparing polyimides having enhanced melt flow properties is described. The process consists of heating a mixture of a high molecular weight poly-(amic acid) or polyimide with a low molecular weight amic acid or imide additive in the range of 0.05 to 15 percent by weight of the additive. The polyimide powders so obtained show improved processability, as evidenced by lower melt viscosity by capillary rheometry. Likewise, films prepared from mixtures of polymers with additives show improved processability with earlier onset of stretching by TMA.

  13. Smog control fuel additives

    SciTech Connect

    Lundby, W.

    1993-06-29

    A method is described of controlling, reducing or eliminating, ozone and related smog resulting from photochemical reactions between ozone and automotive or industrial gases comprising the addition of iodine or compounds of iodine to hydrocarbon-base fuels prior to or during combustion in an amount of about 1 part iodine per 240 to 10,000,000 parts fuel, by weight, to be accomplished by: (a) the addition of these inhibitors during or after the refining or manufacturing process of liquid fuels; (b) the production of these inhibitors for addition into fuel tanks, such as automotive or industrial tanks; or (c) the addition of these inhibitors into combustion chambers of equipment utilizing solid fuels for the purpose of reducing ozone.

  14. Food Additives and Hyperkinesis

    ERIC Educational Resources Information Center

    Wender, Ester H.

    1977-01-01

    The hypothesis that food additives are causally associated with hyperkinesis and learning disabilities in children is reviewed, and available data are summarized. Available from: American Medical Association 535 North Dearborn Street Chicago, Illinois 60610. (JG)

  15. Additional Types of Neuropathy

    MedlinePlus

    ... A A Listen En Español Additional Types of Neuropathy Charcot's Joint Charcot's Joint, also called neuropathic arthropathy, ... can stop bone destruction and aid healing. Cranial Neuropathy Cranial neuropathy affects the 12 pairs of nerves ...

  16. Northern Arizona Volcanoes

    NASA Technical Reports Server (NTRS)

    2006-01-01

    Northern Arizona is best known for the Grand Canyon. Less widely known are the hundreds of geologically young volcanoes, at least one of which buried the homes of local residents. San Francisco Mtn., a truncated stratovolcano at 3887 meters, was once a much taller structure (about 4900 meters) before it exploded some 400,000 years ago a la Mt. St. Helens. The young cinder cone field to its east includes Sunset Crater, that erupted in 1064 and buried Native American homes. This ASTER perspective was created by draping ASTER image data over topographic data from the U.S. Geological Survey National Elevation Data.

    With its 14 spectral bands from the visible to the thermal infrared wavelength region, and its high spatial resolution of 15 to 90 meters (about 50 to 300 feet), ASTER images Earth to map and monitor the changing surface of our planet.

    ASTER is one of five Earth-observing instruments launched December 18, 1999, on NASA's Terra satellite. The instrument was built by Japan's Ministry of Economy, Trade and Industry. A joint U.S./Japan science team is responsible for validation and calibration of the instrument and the data products.

    The broad spectral coverage and high spectral resolution of ASTER provides scientists in numerous disciplines with critical information for surface mapping, and monitoring of dynamic conditions and temporal change. Example applications are: monitoring glacial advances and retreats; monitoring potentially active volcanoes; identifying crop stress; determining cloud morphology and physical properties; wetlands evaluation; thermal pollution monitoring; coral reef degradation; surface temperature mapping of soils and geology; and measuring surface heat balance.

    The U.S. science team is located at NASA's Jet Propulsion Laboratory, Pasadena, Calif. The Terra mission is part of NASA's Science Mission Directorate.

    Size: 20.4 by 24.6 kilometers (12.6 by 15.2 miles) Location: 35.3 degrees North latitude, 111

  17. Detection of IgM antibodies against a recombinant nucleocapsid protein of canine distemper virus in dog sera using a dot-blot assay.

    PubMed

    Barben, G; Stettler, M; Jaggy, A; Vandevelde, M; Zurbriggen, A

    1999-03-01

    A dot-blot assay for the detection of IgM antibodies (ABs) against canine distemper virus (CDV) in canine serum is described. The diagnostic potential of this technique was evaluated by analysing sera from three test groups: (i) specific pathogen-free (SPF) beagle dogs experimentally infected with virulent CDV; (ii) SPF dogs immunized with a combined vaccine containing CDV, and (iii) SPF dogs immunized with a CDV-free vaccine. As antigen for the dot-blot assay we used the recombinant nucleocapsid protein (N protein) of the virulent A75/17 CDV strain. All 12 dogs of group 1, infected with virulent CDV, showed detectable CDV-specific IgM levels in their serum. All dogs of group 2 were also positive for anti-CDV IgM after the first immunization with the CDV-containing vaccine. The four dogs immunized with a CDV-free vaccine (group iii) remained negative throughout the course of the experiment. From these results, we conclude that the IgM detection test, which requires only a single serum sample, is a useful method for diagnosing current or recent CDV infection in CDV-infected or CDV-immunized dogs under experimental conditions. PMID:10216448

  18. Analysis of the insulin receptor gene in noninsulin-dependent diabetes mellitus by denaturing gradient gel blots: A clinical research center study

    SciTech Connect

    Magre, J.; Goldfine, A.B.; Warram, J.H.

    1995-06-01

    We have used a new technique of denaturing gradient gel blotting to determine the prevalence of alterations in the intracellular domain of the insulin receptor in normal individuals and subjects with non-insulin-dependent diabetes mellitus (NIDDM). This method detects DNA sequence differences as restriction fragment melting polymorphisms (RFMP) and is sensitive to changes in sequence at both restriction sites and within the fragments themselves. Using restriction digests with AluI, HaeIII, HinfI, RsaI, Sau3A, and Sau96, 12 RFMPs were found to localize to the region of the {beta}-subunit of the insulin receptor gene. Using exon-specific probes, these RFMPs could be localized to specific regions surrounding individual exons, including exons, 14, 15, 16, 18, 20, and 22. In general, linkage disequilibrium between polymorphisms was inversely related to their distance in the gene structure, although there was a {open_quotes}hot spot{close_quotes} for recombination between exons 19 and 20. No difference in melting temperatures or allele frequency was observed between NIDDM patients and controls. These data indicate that the region of the insulin receptor gene coding for the intracellular portion of the {beta}-subunit is highly polymorphic and that polymorphisms surrounding specific exons can be identified by denaturing gradient gel blotting, but there is no evidence that variation at this locus contributes to NIDDM susceptibility in most individuals. 36 refs., 3 figs., 3 tabs.

  19. Structural characterization of blotting membranes and the influence of membrane parameters for electroblotting and subsequent amino acid sequence analysis of proteins.

    PubMed

    Eckerskorn, C; Lottspeich, F

    1993-09-01

    Various blotting membranes were evaluated and correlated with the efficiency of electroblotting and the performance in the sequencing process. Structural parameters including specific surface area, pore size distribution, pore volumes, and permeabilities of different solvents lead to discrimination of the membranes relative to their accessible surfaces and membrane densities. Protein binding capacities as well as protein recoveries in electroblotting correlate with the specific surface areas. Almost quantitative retention of proteins during electroblotting from gels was obtained for membranes with a high specific surface area and narrow pores (Trans-Blot, Immobilon PSQ, Fluorotrans), whereas membranes with a relatively low specific surface area (Immobilon P, Glassybond) showed reduced recoveries of between 10-20% for the tested proteins. Initial yields and repetitive yields were compared for radioiodinated standard proteins that have been either electroblotted or loaded by direct adsorption. The results showed that the different permeabilities for solutions of the Edman chemistry have a major influence on initial yields. The glass fiber-based membranes with an extremely low flow restriction produce consistently high initial yields independent of the application mode of the protein (spotted or electroblotted) or the application of the membranes into the cartridge (discs or small pieces). In contrast, the polymeric membranes showed decreasing initial yields with increasing membrane density for spotted and electroblotted proteins. Yields varied considerably when the membranes were applied as discs into the cartridge. This effect could be minimized by cutting the membranes into pieces as small as possible, as demonstrated for electroblotted proteins. PMID:8223390

  20. An improved method for western blotting when extracting proteins from mammalian cells cultured on a collagen gel under serum-free conditions.

    PubMed

    Ishihara, Seiichiro; Mizutani, Takeomi; Kawabata, Kazushige; Haga, Hisashi

    2016-01-01

    Western blotting is a widely used method for detection and quantification of specific proteins extracted from mammalian cells. In the conventional method of protein extraction, we found that collagen-containing gels interfered with detection of the p65 protein (one of the subunits in the NF-κB family of proteins) in human lung adenocarcinoma A549 cells cultured on a collagen gel containing serum. In contrast, the collagen gels did not affect detection of the GAPDH protein. Then, we established an improved method for preparation of protein extracts (using trichloroacetic acid fixation and collagenase treatment) from the cells cultured on the collagen gel. Using the improved method, we were able to detect p65 proteins without loss in A549 cells cultured on a collagen gel under serum-free conditions, but we could not detect the proteins if serum was present in cell culture. Thus, using western blotting and serum-free culture conditions, we succeeded in comparing the p65 expression between the cells grown in a plastic dish and cells grown on a collagen gel. PMID:25005915

  1. A Comparison of Antibacterial Activity of Selected Thyme (Thymus) Species by Means of the Dot Blot Test with Direct Bioautographic Detection.

    PubMed

    Orłowska, Marta; Kowalska, Teresa; Sajewicz, Mieczysław; Jesionek, Wioleta; Choma, Irena M; Majer-Dziedzic, Barbara; Szymczak, Grażyna; Waksmundzka-Hajnos, Monika

    2015-01-01

    Bioautography carried out with the aid of thin-layer chromatographic adsorbents can be used to assess antibacterial activity in samples of different origin. It can either be used as a simple and cost-effective detection method applied to a developed chromatogram, or to the dot blot test performed on a chromatographic plate, where total antibacterial activity of a sample is scrutinized. It was an aim of this study to compare antibacterial activity of 18 thyme (Thymus) specimens and species (originating from the same gardening plot and harvested in the same period of time) by means of a dot blot test with direct bioautography. A two-step extraction of herbal material was applied, and at step two the polar fraction of secondary metabolites was obtained under the earlier optimized extraction conditions [methanol-water (27+73, v/v), 130°C]. This fraction was then tested for its antibacterial activity against Bacillus subtilis bacteria. It was established that all investigated extracts exhibited antibacterial activity, yet distinct differences were perceived in the size of the bacterial growth inhibition zones among the compared thyme species. Based on the results obtained, T. citriodorus "golden dwarf" (sample No. 5) and T. marschallianus (sample No. 6) were selected as promising targets for further investigations and possible inclusion in a herbal pharmacopeia, which is an essential scientific novelty of this study. PMID:26268965

  2. Quantifying nuclear p65 as a parameter for NF-κB activation: Correlation between ImageStream cytometry, microscopy, and Western blot.

    PubMed

    Maguire, Orla; Collins, Christine; O'Loughlin, Kieran; Miecznikowski, Jeffrey; Minderman, Hans

    2011-06-01

    The nuclear factor kappa B (NF-κB) pathway, which regulates many cellular processes including proliferation, apoptosis, and survival, has emerged as an important therapeutic target in cancer. Activation of the NF-κB transcription factor is associated with nuclear translocation of the p65 component of the complex. Conventional methods employed to determine nuclear translocation of NF-κB either lack statistical robustness (microscopy) or the ability to discern heterogeneity within the sampled populations (Western blotting and Gel Shift assays). The ImageStream platform combines the high image content information of microscopy with the high throughput and multiparameter analysis of flow cytometry which overcomes the aforementioned limitations of conventional assays. It is demonstrated that ImageStream assessment of receptor-mediated (TNFα) and drug (Daunorubicin, DNR)-induced NF-κB translocation in leukemic cell lines correlates well with microscopy analysis and Western blot analysis. It is further demonstrated that ImageStream cytometry enables quantitative assessment of p65 translocation in immunophenotypically defined subpopulations; and that this assessment is highly reproducible. It is also demonstrated that, quantitatively, the DNR-induced nuclear translocation of NF-κB correlates well with a biological response (apoptosis). We conclude that the ImageStream has the potential to be a powerful tool to evaluate NF-κB /p65 activity as a determinant of response to therapies designed to target aberrant NF-κB signaling activities. PMID:21520400

  3. Additive Manufacturing Infrared Inspection

    NASA Technical Reports Server (NTRS)

    Gaddy, Darrell

    2014-01-01

    Additive manufacturing is a rapid prototyping technology that allows parts to be built in a series of thin layers from plastic, ceramics, and metallics. Metallic additive manufacturing is an emerging form of rapid prototyping that allows complex structures to be built using various metallic powders. Significant time and cost savings have also been observed using the metallic additive manufacturing compared with traditional techniques. Development of the metallic additive manufacturing technology has advanced significantly over the last decade, although many of the techniques to inspect parts made from these processes have not advanced significantly or have limitations. Several external geometry inspection techniques exist such as Coordinate Measurement Machines (CMM), Laser Scanners, Structured Light Scanning Systems, or even traditional calipers and gages. All of the aforementioned techniques are limited to external geometry and contours or must use a contact probe to inspect limited internal dimensions. This presentation will document the development of a process for real-time dimensional inspection technique and digital quality record of the additive manufacturing process using Infrared camera imaging and processing techniques.

  4. Phenylethynyl Containing Reactive Additives

    NASA Technical Reports Server (NTRS)

    Connell, John W. (Inventor); Smith, Joseph G., Jr. (Inventor); Hergenrother, Paul M. (Inventor)

    2002-01-01

    Phenylethynyl containing reactive additives were prepared from aromatic diamine, containing phenylethvnvl groups and various ratios of phthalic anhydride and 4-phenylethynviphthalic anhydride in glacial acetic acid to form the imide in one step or in N-methyl-2-pvrrolidinone to form the amide acid intermediate. The reactive additives were mixed in various amounts (10% to 90%) with oligomers containing either terminal or pendent phenylethynyl groups (or both) to reduce the melt viscosity and thereby enhance processability. Upon thermal cure, the additives react and become chemically incorporated into the matrix and effect an increase in crosslink density relative to that of the host resin. This resultant increase in crosslink density has advantageous consequences on the cured resin properties such as higher glass transition temperature and higher modulus as compared to that of the host resin.

  5. Additives in plastics.

    PubMed Central

    Deanin, R D

    1975-01-01

    The polymers used in plastics are generally harmless. However, they are rarely used in pure form. In almost all commercial plastics, they are "compounded" with monomeric ingredients to improve their processing and end-use performance. In order of total volume used, these monomeric additives may be classified as follows: reinforcing fibers, fillers, and coupling agents; plasticizers; colorants; stabilizers (halogen stabilizers, antioxidants, ultraviolet absorbers, and biological preservatives); processing aids (lubricants, others, and flow controls); flame retardants, peroxides; and antistats. Some information is already available, and much more is needed, on potential toxicity and safe handling of these additives during processing and manufacture of plastics products. PMID:1175566

  6. Biobased lubricant additives

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fully biobased lubricants are those formulated using all biobased ingredients, i.e. biobased base oils and biobased additives. Such formulations provide the maximum environmental, safety, and economic benefits expected from a biobased product. Currently, there are a number of biobased base oils that...

  7. Multifunctional fuel additives

    SciTech Connect

    Baillargeon, D.J.; Cardis, A.B.; Heck, D.B.

    1991-03-26

    This paper discusses a composition comprising a major amount of a liquid hydrocarbyl fuel and a minor low-temperature flow properties improving amount of an additive product of the reaction of a suitable diol and product of a benzophenone tetracarboxylic dianhydride and a long-chain hydrocarbyl aminoalcohol.

  8. Trends in Northern Hemisphere surface cyclone frequency and intensity

    USGS Publications Warehouse

    McCabe, G.J.; Clark, M.P.; Serreze, M.C.

    2001-01-01

    One of the hypothesized effects of global warming from increasing concentrations of greenhouse gases is a change in the frequency and/or intensity of extratropical cyclones. In this study, winter frequencies and intensities of extratropical cyclones in the Northern Hemisphere for the period 1959-97 are examined to determine if identifiable trends are occurring. Results indicate a statistically significant decrease in midlatitude cyclone frequency and a significant increase in high-latitude cyclone frequency. In addition, storm intensity has increased in both the high and midlatitudes. The changes in storm frequency correlate with changes in winter Northern Hemisphere temperature and support hypotheses that global warming may result in a northward shift of storm tracks in the Northern Hemisphere.

  9. Carbon Cycling in Northern Peatlands

    NASA Astrophysics Data System (ADS)

    Schultz, Colin

    2010-11-01

    Northern peatlands span only 3 million square kilometers, about 3% of the terrestrial area of the globe, yet they represent a significant terrestrial sink for carbon dioxide. They are also important emitters of methane, an even more potent greenhouse gas. Despite their substantial role in the global carbon cycle, peatlands are not typically incorporated into global climate models. The AGU Monograph Carbon Cycling in Northern Peatlands, edited by Andrew J. Baird, Lisa R. Belyea, Xavier Comas, A. S. Reeve, and Lee D. Slater, looks at the disproportionate role peatlands play in the global carbon budget. In this interview, Eos talks with Andy Baird, University of Leeds, Leeds, United Kingdom.

  10. Northern Eurasian Heat Waves and Droughts

    NASA Technical Reports Server (NTRS)

    Schubert, Siegfried; Wang, Hailan; Koster, Randal; Suarez, Max; Groisman, Pavel

    2013-01-01

    This article reviews our understanding of the characteristics and causes of northern Eurasian summertime heat waves and droughts. Additional insights into the nature of temperature and precipitation variability in Eurasia on monthly to decadal time scales and into the causes and predictability of the most extreme events are gained from the latest generation of reanalyses and from supplemental simulations with the NASA GEOS-5 AGCM. Key new results are: 1) the identification of the important role of summertime stationary Rossby waves in the development of the leading patterns of monthly Eurasian surface temperature and precipitation variability (including the development of extreme events such as the 2010 Russian heat wave), 2) an assessment of the mean temperature and precipitation changes that have occurred over northern Eurasia in the last three decades and their connections to decadal variability and global trends in SST, and 3) the quantification (via a case study) of the predictability of the most extreme simulated heat wave/drought events, with some focus on the role of soil moisture in the development and maintenance of such events. A literature survey indicates a general consensus that the future holds an enhanced probability of heat waves across northern Eurasia, while there is less agreement regarding future drought, reflecting a greater uncertainty in soil moisture and precipitation projections. Substantial uncertainties remain in our understanding of heat waves and drought, including the nature of the interactions between the short-term atmospheric variability associated with such extremes and the longer-term variability and trends associated with soil moisture feedbacks, SST anomalies, and an overall warming world.

  11. Neotectonics in the northern equatorial Brazilian margin

    NASA Astrophysics Data System (ADS)

    Rossetti, Dilce F.; Souza, Lena S. B.; Prado, Renato; Elis, Vagner R.

    2012-08-01

    An increasing volume of publications has addressed the role of tectonics in inland areas of northern Brazil during the Neogene and Quaternary, despite its location in a passive margin. Hence, northern South America plate in this time interval might have not been as passive as usually regarded. This proposal needs further support, particularly including field data. In this work, we applied an integrated approach to reveal tectonic structures in Miocene and late Quaternary strata in a coastal area of the Amazonas lowland. The investigation, undertaken in Marajó Island, mouth of the Amazonas River, consisted of shallow sub-surface geophysical data including vertical electric sounding and ground penetrating radar. These methods were combined with morphostructural analysis and sedimentological/stratigraphic data from shallow cores and a few outcrops. The results revealed two stratigraphic units, a lower one with Miocene age, and an upper one of Late Pleistocene-Holocene age. An abundance of faults and folds were recorded in the Miocene deposits and, to a minor extent, in overlying Late Pleistocene-Holocene strata. In addition to characterize these structures, we discuss their origin, considering three potential mechanisms: Andean tectonics, gravity tectonics related to sediment loading in the Amazon Fan, and rifting at the continental margin. Amongst these hypotheses, the most likely is that the faults and folds recorded in Marajó Island reflect tectonics associated with the history of continental rifting that gave rise to the South Atlantic Ocean. This study supports sediment deposition influenced by transpression and transtension associated with strike-slip divergence along the northern Equatorial Brazilian margin in the Miocene and Late Pleistocene-Holocene. This work records tectonic evidence only for the uppermost few ten of meters of this sedimentary succession. However, available geological data indicate a thickness of up to 6 km, which is remarkably thick for

  12. A simple, inexpensive, robust and sensitive dot-blot assay for equal detection of the nonstructural-1 glycoprotein of all dengue virus serotypes

    PubMed Central

    2013-01-01

    Background Detection of dengue virus (DENV) soluble/excreted (s/e) form of the nonstructural-1 (NS1) glycoprotein in patient acute-phase sera is ideal for diagnosis. The commercially-available detection assays are, however, too expensive for routine use and have low specificity, particularly for the s/e NS1 glycoprotein of DENV-2 and DENV-4, which are important causes of lethal human disease worldwide. Methods Mouse monoclonal antibodies (MAbs) were generated and screened against s/e NS1 glycoprotein purified from each DENV serotype to obtain those that reacted equally with each serotype, but not with yellow fever virus (YFV) s/e NS1 glycoprotein or human serum proteins. One MAb, MAb 2C4.6, was further tested against these DENV glycoproteins in human sera using simple, peroxidase-labelled secondary antibody/substrate-developed dot-blot assays. Results Optimal quenching of endogenous human serum peroxidases was attained using 3% H2O2 in H20 for 5 min. MAb 2C4.6 showed an acceptable detection sensitivity of < 32 ng/ml for the s/e NS1 glycoprotein of each DENV serotype but did not cross-react with the YFV s/e NS1 glycoprotein or human serum proteins. By contrast, the LX1 epitope-specific MAb, 3D1.4, showed similar detection sensitivity against only the DENV-1 NS1 glycoprotein, consistent with results from commercial DENV s/e NS1 glycoprotein detection assays. DENV s/e NS1 glycoproteins were stable in human sera after drying on the nitrocellulose membranes and storage for one month at ambient temperature (28°C) before being processed. The total assay time was reduced to 3 h without any loss of detection sensitivity. This dot-blot format was ideal for the circulating immune complex disruption step, which is required for increased DENV s/e NS1 glycoprotein detection. Conclusions This is the first study to determine the detection sensitivity of MAbs against known concentrations of s/e NS1 glycoprotein from each DENV serotype. The preparation of patient serum samples for

  13. Titan's Stratospheric Condensibles at High Northern Latitudes During Northern Winter

    NASA Technical Reports Server (NTRS)

    Anderson, Carrie; Samuelson, R.; Achterberg, R.

    2012-01-01

    The Infrared Interferometer Spectrometer (IRIS) instrument on board Voyager 1 caught the first glimpse of an unidentified particulate feature in Titan's stratosphere that spectrally peaks at 221 per centimeter. Until recently, this feature that we have termed 'the haystack,' has been seen persistently at high northern latitudes with the Composite Infrared Spectrometer (CIRS) instrument onboard Cassini, The strength of the haystack emission feature diminishes rapidly with season, becoming drastically reduced at high northern latitudes, as Titan transitions from northern winter into spring, In contrast to IRIS whose shortest wavenumber was 200 per centimeter, CIRS extends down to 10 per centimeter, thus revealing an entirely unexplored spectral region in which nitrile ices have numerous broad lattice vibration features, Unlike the haystack, which is only found at high northern latitudes during northern winter/early northern spring, this geometrically thin nitrile cloud pervades Titan's lower stratosphere, spectrally peaking at 160 per centimeter, and is almost global in extent spanning latitudes 85 N to 600 S, The inference of nitrile ices are consistent with the highly restricted altitude ranges over which these features are observed, and appear to be dominated by a mixture of HCN and HC3N, The narrow range in altitude over which the nitrile ices extend is unlike the haystack, whose vertical distribution is significantly broader, spanning roughly 70 kilometers in altitude in Titan's lower stratosphere, The nitrile clouds that CIRS observes are located in a dynamically stable region of Titan's atmosphere, whereas CH4 clouds, which ordinarily form in the troposphere, form in a more dynamically unstable region, where convective cloud systems tend to occur. In the unusual situation where Titan's tropopause cools significantly from the HASI 70.5K temperature minimum, CH4 should condense in Titan's lower stratosphere, just like the aforementioned nitrile clouds, although

  14. Boron addition to alloys

    SciTech Connect

    Coad, B. C.

    1985-08-20

    A process for addition of boron to an alloy which involves forming a melt of the alloy and a reactive metal, selected from the group consisting of aluminum, titanium, zirconium and mixtures thereof to the melt, maintaining the resulting reactive mixture in the molten state and reacting the boric oxide with the reactive metal to convert at least a portion of the boric oxide to boron which dissolves in the resulting melt, and to convert at least portion of the reactive metal to the reactive metal oxide, which oxide remains with the resulting melt, and pouring the resulting melt into a gas stream to form a first atomized powder which is subsequently remelted with further addition of boric oxide, re-atomized, and thus reprocessed to convert essentially all the reactive metal to metal oxide to produce a powdered alloy containing specified amounts of boron.

  15. Tackifier for addition polyimides

    NASA Technical Reports Server (NTRS)

    Butler, J. M.; St.clair, T. L.

    1980-01-01

    A modification to the addition polyimide, LaRC-160, was prepared to improve tack and drape and increase prepeg out-time. The essentially solventless, high viscosity laminating resin is synthesized from low cost liquid monomers. The modified version takes advantage of a reactive, liquid plasticizer which is used in place of solvent and helps solve a major problem of maintaining good prepeg tack and drape, or the ability of the prepeg to adhere to adjacent plies and conform to a desired shape during the lay up process. This alternate solventless approach allows both longer life of the polymer prepeg and the processing of low void laminates. This approach appears to be applicable to all addition polyimide systems.

  16. Vinyl capped addition polyimides

    NASA Technical Reports Server (NTRS)

    Vannucci, Raymond D. (Inventor); Malarik, Diane C. (Inventor); Delvigs, Peter (Inventor)

    1991-01-01

    Polyimide resins (PMR) are generally useful where high strength and temperature capabilities are required (at temperatures up to about 700 F). Polyimide resins are particularly useful in applications such as jet engine compressor components, for example, blades, vanes, air seals, air splitters, and engine casing parts. Aromatic vinyl capped addition polyimides are obtained by reacting a diamine, an ester of tetracarboxylic acid, and an aromatic vinyl compound. Low void materials with improved oxidative stability when exposed to 700 F air may be fabricated as fiber reinforced high molecular weight capped polyimide composites. The aromatic vinyl capped polyimides are provided with a more aromatic nature and are more thermally stable than highly aliphatic, norbornenyl-type end-capped polyimides employed in PMR resins. The substitution of aromatic vinyl end-caps for norbornenyl end-caps in addition polyimides results in polymers with improved oxidative stability.

  17. [Biologically active food additives].

    PubMed

    Velichko, M A; Shevchenko, V P

    1998-07-01

    More than half out of 40 projects for the medical science development by the year of 2000 have been connected with the bio-active edible additives that are called "the food of XXI century", non-pharmacological means for many diseases. Most of these additives--nutricevtics and parapharmacevtics--are intended for the enrichment of food rations for the sick or healthy people. The ecologicaly safest and most effective are combined domestic adaptogens with immuno-modulating and antioxidating action that give anabolic and stimulating effect,--"leveton", "phytoton" and "adapton". The MKTs-229 tablets are residue discharge means. For atherosclerosis and general adiposis they recommend "tsar tablets" and "aiconol (ikhtien)"--on the base of cod-liver oil or "splat" made out of seaweed (algae). All these preparations have been clinically tested and received hygiene certificates from the Institute of Dietology of the Russian Academy of Medical Science. PMID:9752776

  18. Electrophilic addition of astatine

    SciTech Connect

    Norseev, Yu.V.; Vasaros, L.; Nhan, D.D.; Huan, N.K.

    1988-03-01

    It has been shown for the first time that astatine is capable of undergoing addition reactions to unsaturated hydrocarbons. A new compound of astatine, viz., ethylene astatohydrin, has been obtained, and its retention numbers of squalane, Apiezon, and tricresyl phosphate have been found. The influence of various factors on the formation of ethylene astatohydrin has been studied. It has been concluded on the basis of the results obtained that the univalent cations of astatine in an acidic medium is protonated hypoastatous acid.

  19. Hydrocarbon fuel additive

    SciTech Connect

    Ambrogio, S.

    1989-02-28

    This patent describes the method of fuel storage or combustion, wherein the fuel supply contains small amounts of water, the step of adding to the fuel supply an additive comprising a blend of a hydrophilic agent chosen from the group of ethylene glycol, n-butyl alcohol, and cellosolve in the range of 22-37% by weight; ethoxylated nonylphenol in the range of 26-35% by weight; nonylphenol polyethylene glycol ether in the range of 32-43% by weight.

  20. Functional Generalized Additive Models.

    PubMed

    McLean, Mathew W; Hooker, Giles; Staicu, Ana-Maria; Scheipl, Fabian; Ruppert, David

    2014-01-01

    We introduce the functional generalized additive model (FGAM), a novel regression model for association studies between a scalar response and a functional predictor. We model the link-transformed mean response as the integral with respect to t of F{X(t), t} where F(·,·) is an unknown regression function and X(t) is a functional covariate. Rather than having an additive model in a finite number of principal components as in Müller and Yao (2008), our model incorporates the functional predictor directly and thus our model can be viewed as the natural functional extension of generalized additive models. We estimate F(·,·) using tensor-product B-splines with roughness penalties. A pointwise quantile transformation of the functional predictor is also considered to ensure each tensor-product B-spline has observed data on its support. The methods are evaluated using simulated data and their predictive performance is compared with other competing scalar-on-function regression alternatives. We illustrate the usefulness of our approach through an application to brain tractography, where X(t) is a signal from diffusion tensor imaging at position, t, along a tract in the brain. In one example, the response is disease-status (case or control) and in a second example, it is the score on a cognitive test. R code for performing the simulations and fitting the FGAM can be found in supplemental materials available online. PMID:24729671

  1. High rate of missed HIV infections in individuals with indeterminate or negative HIV western blots based on current HIV testing algorithm in China.

    PubMed

    Liu, Man-Qing; Zhu, Ze-Rong; Kong, Wen-Hua; Tang, Li; Peng, Jin-Song; Wang, Xia; Xu, Jun; Schilling, Robert F; Cai, Thomas; Zhou, Wang

    2016-08-01

    It remains unclear if China's current HIV antibody testing algorithm misses a substantial number of HIV infected individuals. Of 196 specimens with indeterminate or negative results on HIV western blot (WB) retrospectively examined by HIV-1 nucleic acid test (NAT), 67.57% (75/111) of indeterminate WB samples, and 16.47% (14/85) of negative WB samples were identified as NAT positive. HIV-1 loads in negative WB samples were significantly higher than those in indeterminate WB samples. Notably, 86.67% (13/15) of samples with negative WB and double positive immunoassay results were NAT positive. The rate of HIV-1 infections missed by China's current HIV testing algorithm is unacceptably high. Thus, China should consider using NAT or integrating fourth generation ELISA into current only antibodies-based HIV confirmation. J. Med. Virol. 88:1462-1466, 2016. © 2016 Wiley Periodicals, Inc. PMID:26856240

  2. From field to gel blot: teaching a holistic view of developmental phenomena to undergraduate biology students at the University of Tokyo.

    PubMed

    Ariizumi, Takashi; Asashima, Makoto

    2003-01-01

    We present here an outline of the lectures and laboratory exercises for undergraduate developmental biology students at the University of Tokyo. The main aim of our course is to help students fill the gap between natural history, classical embryology and molecular developmental biology. To achieve this aim, we take up various topics in the lectures, from fertilization and early development to developmental engineering. Our laboratory exercises begin with an introduction to the natural history of the organism. The entire class and the instructors collect newts in the field and discuss features of their mating behavior and so on. In the laboratory, students are absorbed by exercises such as a lampbrush chromosome preparation and an in vitro beating heart induction. After that, students choose their own research projects for which they will employ both classical embryological and modern molecular biological techniques. At the end of our course, the connectivity principle from field to gel blot will be part of the students' understanding. PMID:12705655

  3. Detection of Ca(2+)-binding proteins by electrophoretic migration in the presence of Ca2+ combined with 45Ca2+ overlay of protein blots

    SciTech Connect

    Garrigos, M.; Deschamps, S.; Viel, A.; Lund, S.; Champeil, P.; Moller, J.V.; le Maire, M. , Gif-sur-Yvette )

    1991-04-01

    When high affinity Ca(2+)-binding proteins like calmodulin, or proteins with a high Ca(2+)-binding capacity like calsequestrin, underwent sodium dodecyl sulfate-gel electrophoresis in Laemmli systems, their electrophoretic migration rates were much higher in gels containing 1 mM Ca2+ than in gels containing ethylene glycol bis(beta-aminoethyl ether) N,N{prime}-tetraacetic acid (EGTA). Replacement of EGTA by Ca2+ in the gel, combined with the blotting of electrophoretically separated proteins on polyvinylidene difluoride membranes and subsequent 45Ca2+ overlay, proved a very effective means of detecting Ca(2+)-binding proteins. This combined approach is important since artifacts occur in both techniques when used separately. We found that the usual procedure of adding Ca2+ to the sample before electrophoresis without including it in the gel itself permitted the detection of only very high affinity Ca(2+)-binding proteins.

  4. Investigation of cross-reactions against Trichinella spiralis antigens by enzyme-linked immunosorbent assay and enzyme-linked immunoelectrotransfer blot assay in patients with various diseases.

    PubMed Central

    De-la-Rosa, J L; Alcantara, P; Correa, D

    1995-01-01

    Data regarding cross-reactions against Trichinella spiralis in humans are scarce and controversial. For this reason, we tested serum samples from patients with typhoid fever, brucellosis, toxoplasmosis, amoebiasis, cysticercosis, trichocephaliasis, ascariasis, and onchocerciasis against an antigenic extract of T. spiralis infective larvae in an enzyme-linked immunosorbent assay (ELISA) and an enzyme-linked immunoelectrotransfer blot (EITB) assay. All except one serum sample from the group of patients with onchocerciasis were negative in the ELISA; in the EITB assay, only faint bands were observed with the samples from patients with onchocerciasis and ascariasis and negative results were obtained with the samples from patients with other diseases. In conclusion, cross-reactions were found only in the groups of patients with other nematode infections and were of very low magnitude, most of them virtually negative. PMID:7719905

  5. Application of glyco-blotting for identification of structures of polysaccharides causing membrane fouling in a pilot-scale membrane bioreactor treating municipal wastewater.

    PubMed

    Kimura, Katsuki; Nishimura, Shin-Ichiro; Miyoshi, Risho; Hoque, Asiful; Miyoshi, Taro; Watanabe, Yoshimasa

    2015-03-01

    A new approach for the analysis of polysaccharides in membrane bioreactor (MBR) is proposed in this study. Enrichment of polysaccharides by glyco-blotting, in which polysaccharides are specifically collected via interactions between the aldehydes in the polysaccharides and aminooxy groups on glycoblotting beads, enabled MALDI-TOF/MS analysis at a high resolution. Structures of polysaccharides extracted from fouled membranes used in a pilot-scale MBR treating municipal wastewater and those in the supernatant of the mixed liquor suspension in the MBR were investigated. It was found that the overlap between polysaccharides found in the supernatants and those extracted from the fouled membrane was rather limited, suggesting that polysaccharides that dominate in supernatants may not be important in membrane fouling in MBRs. Analysis using a bacterial carbohydrate database suggested that capsular polysaccharides (CPS) and/or lipo-polysaccharides (LPS) produced by gram-negative bacteria are key players in the evolution of membrane fouling in MBRs. PMID:25544495

  6. Serum detection of IgG antibodies against Demodex canis by western blot in healthy dogs and dogs with juvenile generalized demodicosis.

    PubMed

    Ravera, Ivan; Ferreira, Diana; Gallego, Laia Solano; Bardagí, Mar; Ferrer, Lluís

    2015-08-01

    The aim of this study was to investigate the presence of canine immunoglobulins (Ig) G against Demodex proteins in the sera of healthy dogs and of dogs with juvenile generalized demodicosis (CanJGD) with or without secondary pyoderma. Demodex mites were collected from dogs with CanJGD. Protein concentration was measured and a western blot technique was performed. Pooled sera from healthy dogs reacted mainly with antigen bands ranging from 55 to 72 kDa. Pooled sera from dogs with CanJGD without secondary pyoderma reacted either with 10 kDa antigen band or 55 to 72 kDa bands. Pooled sera from dogs with CanJGD with secondary pyoderma reacted only with a 10 kDa antigen band. The results of this study suggest that both healthy dogs and dogs with CanJGD develop a humoral response against different proteins of Demodex canis. PMID:26267107

  7. Localization and Distribution of 'Candidatus Liberibacter asiaticus' in Citrus and Periwinkle by Direct Tissue Blot Immuno Assay with an Anti-OmpA Polyclonal Antibody.

    PubMed

    Ding, Fang; Duan, Yongping; Paul, Cristina; Brlansky, Ronald H; Hartung, John S

    2015-01-01

    'Candidatus Liberibacter asiaticus' (CaLas), a non-cultured member of the α-proteobacteria, is the causal agent of citrus Huanglongbing (HLB). Due to the difficulties of in vitro culture, antibodies against CaLas have not been widely used in studies of this pathogen. We have used an anti-OmpA polyclonal antibody based direct tissue blot immunoassay to localize CaLas in different citrus tissues and in periwinkle leaves. In citrus petioles, CaLas was unevenly distributed in the phloem sieve tubes, and tended to colonize in phloem sieve tubes on the underside of petioles in preference to the upper side of petioles. Both the leaf abscission zone and the junction of the petiole and leaf midrib had fewer CaLas bacteria compared to the main portions of the petiole and the midribs. Colonies of CaLas in phloem sieve tubes were more frequently found in stems with symptomatic leaves than in stems with asymptomatic leaves with an uneven distribution pattern. In serial sections taken from the receptacle to the peduncle, more CaLas were observed in the peduncle sections adjacent to the stem. In seed, CaLas was located in the seed coat. Many fewer CaLas were found in the roots, as compared to the seeds and petioles when samples were collected from trees with obvious foliar symptoms. The direct tissue blot immuno assay was adapted to whole periwinkle leaves infected by CaLas. The pathogen was distributed throughout the lateral veins and the results were correlated with results of qPCR. Our data provide direct spatial and anatomical information for CaLas in planta. This simple and scalable method may facilitate the future research on the interaction of CaLas and host plant. PMID:25946013

  8. A Secondary Antibody-Detecting Molecular Weight Marker with Mouse and Rabbit IgG Fc Linear Epitopes for Western Blot Analysis

    PubMed Central

    Cheng, Ta-Chun; Tung, Yi-Ching; Chu, Pei-Yu; Chuang, Chih-Hung; Hsieh, Yuan-Chin; Huang, Chien-Chiao; Wang, Yeng-Tseng; Kao, Chien-Han; Roffler, Steve R.; Cheng, Tian-Lu

    2016-01-01

    Molecular weight markers that can tolerate denaturing conditions and be auto-detected by secondary antibodies offer great efficacy and convenience for Western Blotting. Here, we describe M&R LE protein markers which contain linear epitopes derived from the heavy chain constant regions of mouse and rabbit immunoglobulin G (IgG Fc LE). These markers can be directly recognized and stained by a wide range of anti-mouse and anti-rabbit secondary antibodies. We selected three mouse (M1, M2 and M3) linear IgG1 and three rabbit (R1, R2 and R3) linear IgG heavy chain epitope candidates based on their respective crystal structures. Western blot analysis indicated that M2 and R2 linear epitopes are effectively recognized by anti-mouse and anti-rabbit secondary antibodies, respectively. We fused the M2 and R2 epitopes (M&R LE) and incorporated the polypeptide in a range of 15–120 kDa auto-detecting markers (M&R LE protein marker). The M&R LE protein marker can be auto-detected by anti-mouse and anti-rabbit IgG secondary antibodies in standard immunoblots. Linear regression analysis of the M&R LE protein marker plotted as gel mobility versus the log of the marker molecular weights revealed good linearity with a correlation coefficient R2 value of 0.9965, indicating that the M&R LE protein marker displays high accuracy for determining protein molecular weights. This accurate, regular and auto-detected M&R LE protein marker may provide a simple, efficient and economical tool for protein analysis. PMID:27494183

  9. Western blot analysis of BK channel β1‐subunit expression should be interpreted cautiously when using commercially available antibodies

    PubMed Central

    Bhattarai, Yogesh; Fernandes, Roxanne; Kadrofske, Mark M.; Lockwood, Lizbeth R.; Galligan, James J.; Xu, Hui

    2014-01-01

    Abstract Large conductance Ca2+‐activated K+ (BK) channels consist of pore‐forming α‐ and accessory β‐subunits. There are four β‐subunit subtypes (β1–β4), BK β1‐subunit is specific for smooth muscle cells (SMC). Reduced BK β1‐subunit expression is associated with SMC dysfunction in animal models of human disease, because downregulation of BK β1‐subunit reduces channel activity and increases SMC contractility. Several anti‐BK β1‐subunit antibodies are commercially available; however, the specificity of most antibodies has not been tested or confirmed in the tissues from BK β1‐subunit knockout (KO) mice. In this study, we tested the specificity and sensitivity of six commercially available antibodies from five manufacturers. We performed western blot analysis on BK β1‐subunit enriched tissues (mesenteric arteries and colons) and non‐SM tissue (cortex of kidney) from wild‐type (WT) and BK β1‐KO mice. We found that antibodies either detected protein bands of the appropriate molecular weight in tissues from both WT and BK β1‐KO mice or failed to detect protein bands at the appropriate molecular weight in tissues from WT mice, suggesting that these antibodies may lack specificity for the BK β1‐subunit. The absence of BK β1‐subunit mRNA expression in arteries, colons, and kidneys from BK β1‐KO mice was confirmed by RT‐PCR analysis. We conclude that these commercially available antibodies might not be reliable tools for studying BK β1‐subunit expression in murine tissues under the denaturing conditions that we have used. Data obtained using commercially available antibodies should be interpreted cautiously. Our studies underscore the importance of proper negative controls in western blot analyses. PMID:25355855

  10. Localization and Distribution of 'Candidatus Liberibacter asiaticus’ in Citrus and Periwinkle by Direct Tissue Blot Immuno Assay with an Anti-OmpA Polyclonal Antibody

    PubMed Central

    Ding, Fang; Duan, Yongping; Paul, Cristina; Brlansky, Ronald H.; Hartung, John S.

    2015-01-01

    ‘Candidatus Liberibacter asiaticus’ (CaLas), a non-cultured member of the α-proteobacteria, is the causal agent of citrus Huanglongbing (HLB). Due to the difficulties of in vitro culture, antibodies against CaLas have not been widely used in studies of this pathogen. We have used an anti-OmpA polyclonal antibody based direct tissue blot immunoassay to localize CaLas in different citrus tissues and in periwinkle leaves. In citrus petioles, CaLas was unevenly distributed in the phloem sieve tubes, and tended to colonize in phloem sieve tubes on the underside of petioles in preference to the upper side of petioles. Both the leaf abscission zone and the junction of the petiole and leaf midrib had fewer CaLas bacteria compared to the main portions of the petiole and the midribs. Colonies of CaLas in phloem sieve tubes were more frequently found in stems with symptomatic leaves than in stems with asymptomatic leaves with an uneven distribution pattern. In serial sections taken from the receptacle to the peduncle, more CaLas were observed in the peduncle sections adjacent to the stem. In seed, CaLas was located in the seed coat. Many fewer CaLas were found in the roots, as compared to the seeds and petioles when samples were collected from trees with obvious foliar symptoms. The direct tissue blot immuno assay was adapted to whole periwinkle leaves infected by CaLas. The pathogen was distributed throughout the lateral veins and the results were correlated with results of qPCR. Our data provide direct spatial and anatomical information for CaLas in planta. This simple and scalable method may facilitate the future research on the interaction of CaLas and host plant. PMID:25946013

  11. Northern New Mexico regional airport market feasibility

    SciTech Connect

    Drake, R.H.; Williams, D.S.

    1998-06-01

    This report is about the market for airline travel in northern New Mexico. Interest in developing a northern New Mexico regional airport has periodically surfaced for a number of years. The New Mexico State Legislature passed a memorial during the 1998 Second Session calling for the conduct of a study to determine the feasibility of building a new regional airport in NNM. This report is a study of the passenger market feasibility of such an airport. In addition to commercial passenger market feasibility, there are other feasibility issues dealing with siting, environmental impact, noise, economic impact, intermodal transportation integration, region-wide transportation services, airport engineering requirements, and others. These other feasibility issues are not analyzed in any depth in this report although none were discovered to be show-stoppers as a by-product of the authors doing research on the passenger market itself. Preceding the need for a detailed study of these other issues is the determination of the basic market need for an airport with regular commercial airline service in the first place. This report is restricted to an in-depth look at the market for commercial passenger air service in NNM. 20 figs., 8 tabs.

  12. Opsin gene repertoires in northern archaic hominids.

    PubMed

    Taylor, John S; Reimchen, Thomas E

    2016-08-01

    The Neanderthals' northern distribution, hunting techniques, and orbit breadths suggest that they were more active in dim light than modern humans. We surveyed visual opsin genes from four Neanderthals and two other archaic hominids to see if they provided additional support for this hypothesis. This analysis was motivated by the observation that alleles responsible for anomalous trichromacy in humans are more common in northern latitudes, by data suggesting that these variants might enhance vision in mesopic conditions, and by the observation that dim light active species often have fewer opsin genes than diurnal relatives. We also looked for evidence of convergent amino acid substitutions in Neanderthal opsins and orthologs from crepuscular or nocturnal species. The Altai Neanderthal, the Denisovan, and the Ust'-Ishim early modern human had opsin genes that encoded proteins identical to orthologs in the human reference genome. Opsins from the Vindija Cave Neanderthals (three females) had many nonsynonymous substitutions, including several predicted to influence colour vision (e.g., stop codons). However, the functional implications of these observations were difficult to assess, given that "control" loci, where no substitutions were expected, differed from humans to the same extent. This left unresolved the test for colour vision deficiencies in Vindija Cave Neanderthals. PMID:27463216

  13. NORTHERN PUGET SOUND MARINE MAMMALS

    EPA Science Inventory

    A baseline study of the marine mammals of northern Puget Sound and the Strait of Juan de Fuca was undertaken from November 1977 to September 1979 emphasizing certain aspects of the biology of the harbor seal, which is the most abundant marine mammal in this area. The local abunda...

  14. "Subtractive" Bilingualism in Northern Belize.

    ERIC Educational Resources Information Center

    Rubinstein, Robert A.

    "Subtractive" bilingualism in Northern Belize is analyzed based on an extension of a model by Wallace Lambert. The impact of English language instruction on Spanish speaking children in Corozal Town, the northernmost urban center in the British colony of Belize, Central America, is described. This description extends an earlier account of…

  15. NUMA: A Northern Paiute History.

    ERIC Educational Resources Information Center

    Inter-Tribal Council of Nevada, Reno.

    One in a series of four histories of native Nevadans, this volume presents the story of the Northern Paiute people, or Numa, who lived, hunted, and travelled in the Great Basin area which occupies one-third of present day Nevada and parts of Oregon, Idaho, and California. Based on interviews with tribal elders and research conducted at numerous…

  16. Spatial profiles of hsp70 proteins in Asian clam (Potamocorbula amurensis) in northern San Francisco Bay may be linked to natural rather than anthropogenic stressors.

    PubMed

    Werner, I; Hinton, D E

    2000-01-01

    Multi-year investigations in northern San Francisco Bay by United States Geological Survey have linked reduced condition indices in populations of Asian clam (Potamocorbula amurensis) with elevated cadmium tissue concentrations. Our study seeks to determine whether levels of hsp70 proteins in P. amurensis can be correlated with these findings, and/or are related to histopathologic alterations and concentrations of metallothionein-like proteins. Here we present our results on stress proteins in clams collected monthly from four field stations between July 1996 and January 1998. In addition, animals were exposed in the laboratory to a range of salinities. Stress proteins were analyzed by Western blotting using monoclonal antibodies. Hsp70 protein levels in field-collected clams were significantly higher at the seaward (high salinity/low cadmium) stations (12.5, 8.1) than at the landward (low salinity/high cadmium) stations (6.1, 4.1). Laboratory studies showed that clams exposed to 0.1 ppt salinity had markedly lower hsp70 levels than clams exposed to higher salinities. In view of our previous laboratory studies showing that cadmium induces hsp70 in P. amurensis, our present results indicate that reduced hsp70 protein levels in field-collected clams may be linked to salinity effects rather than cadmium tissue concentrations. PMID:11460722

  17. Siloxane containing addition polyimides

    NASA Technical Reports Server (NTRS)

    Maudgal, S.; St. Clair, T. L.

    1984-01-01

    Addition polyimide oligomers have been synthesized from bis(gamma-aminopropyl) tetramethyldisiloxane and 3, 3', 4, 4'-benzophenonetetracarboxylic dianhydride using a variety of latent crosslinking groups as endcappers. The prepolymers were isolated and characterized for solubility (in amide, chlorinated and ether solvents), melt flow and cure properties. The most promising systems, maleimide and acetylene terminated prepolymers, were selected for detailed study. Graphite cloth reinforced composites were prepared and properties compared with those of graphite/Kerimid 601, a commercially available bismaleimide. Mixtures of the maleimide terminated system with Kerimid 601, in varying proportions, were also studied.

  18. Oil additive process

    SciTech Connect

    Bishop, H.

    1988-10-18

    This patent describes a method of making an additive comprising: (a) adding 2 parts by volume of 3% sodium hypochlorite to 45 parts by volume of diesel oil fuel to form a sulphur free fuel, (b) removing all water and foreign matter formed by the sodium hypochlorite, (c) blending 30 parts by volume of 24% lead naphthanate with 15 parts by volume of the sulphur free fuel, 15 parts by volume of light-weight material oil to form a blended mixture, and (d) heating the blended mixture slowly and uniformly to 152F.

  19. Performance Boosting Additive

    NASA Technical Reports Server (NTRS)

    1999-01-01

    Mainstream Engineering Corporation was awarded Phase I and Phase II contracts from Goddard Space Flight Center's Small Business Innovation Research (SBIR) program in early 1990. With support from the SBIR program, Mainstream Engineering Corporation has developed a unique low cost additive, QwikBoost (TM), that increases the performance of air conditioners, heat pumps, refrigerators, and freezers. Because of the energy and environmental benefits of QwikBoost, Mainstream received the Tibbetts Award at a White House Ceremony on October 16, 1997. QwikBoost was introduced at the 1998 International Air Conditioning, Heating, and Refrigeration Exposition. QwikBoost is packaged in a handy 3-ounce can (pressurized with R-134a) and will be available for automotive air conditioning systems in summer 1998.

  20. Sewage sludge additive

    NASA Technical Reports Server (NTRS)

    Kalvinskas, J. J.; Mueller, W. A.; Ingham, J. D. (Inventor)

    1980-01-01

    The additive is for a raw sewage treatment process of the type where settling tanks are used for the purpose of permitting the suspended matter in the raw sewage to be settled as well as to permit adsorption of the dissolved contaminants in the water of the sewage. The sludge, which settles down to the bottom of the settling tank is extracted, pyrolyzed and activated to form activated carbon and ash which is mixed with the sewage prior to its introduction into the settling tank. The sludge does not provide all of the activated carbon and ash required for adequate treatment of the raw sewage. It is necessary to add carbon to the process and instead of expensive commercial carbon, coal is used to provide the carbon supplement.

  1. Perspectives on Additive Manufacturing

    NASA Astrophysics Data System (ADS)

    Bourell, David L.

    2016-07-01

    Additive manufacturing (AM) has skyrocketed in visibility commercially and in the public sector. This article describes the development of this field from early layered manufacturing approaches of photosculpture, topography, and material deposition. Certain precursors to modern AM processes are also briefly described. The growth of the field over the last 30 years is presented. Included is the standard delineation of AM technologies into seven broad categories. The economics of AM part generation is considered, and the impacts of the economics on application sectors are described. On the basis of current trends, the future outlook will include a convergence of AM fabricators, mass-produced AM fabricators, enabling of topology optimization designs, and specialization in the AM legal arena. Long-term developments with huge impact are organ printing and volume-based printing.

  2. New addition curing polyimides

    NASA Technical Reports Server (NTRS)

    Frimer, Aryeh A.; Cavano, Paul

    1991-01-01

    In an attempt to improve the thermal-oxidative stability (TOS) of PMR-type polymers, the use of 1,4-phenylenebis (phenylmaleic anhydride) PPMA, was evaluated. Two series of nadic end-capped addition curing polyimides were prepared by imidizing PPMA with either 4,4'-methylene dianiline or p-phenylenediamine. The first resulted in improved solubility and increased resin flow while the latter yielded a compression molded neat resin sample with a T(sub g) of 408 C, close to 70 C higher than PME-15. The performance of these materials in long term weight loss studies was below that of PMR-15, independent of post-cure conditions. These results can be rationalized in terms of the thermal lability of the pendant phenyl groups and the incomplete imidization of the sterically congested PPMA. The preparation of model compounds as well as future research directions are discussed.

  3. Land use of northern megalopolis

    NASA Technical Reports Server (NTRS)

    Simpson, R. B.; Lindgren, D. T.

    1973-01-01

    The major objective is to map and digitize the land use of northern megalopolis, the states of Massachusetts, Connecticut, and Rhode Island, and to evaluate ERTS as a planning tool for megalopolitan areas. The southern New England region provides a good test ERTS's capabilities because of its complex landscape. Not only are there great differences in the degree of urban development, but in relief and vegetative cover as well.

  4. The northern Egyptian continental margin

    NASA Astrophysics Data System (ADS)

    Badawy, Ahmed; Mohamed, Gad; Omar, Khaled; Farid, Walid

    2015-01-01

    Africa displays a variety of continental margin structures, tectonics and sedimentary records. The northern Egyptian continental margin represents the NE portion of the North African passive continental margin. Economically, this region is of great importance as a very rich and productive hydrocarbon zone in Egypt. Moreover, it is characterized by remarkable tectonic setting accompanied by active tectonic processes from the old Tethys to recent Mediterranean. In this article, seismicity of the northern Egyptian continental margin has been re-evaluated for more than 100-years and the source parameters of three recent earthquakes (October 2012, January 2013 and July 2013) have been estimated. Moment tensor inversions of 19th October 2012 and 17th January 2013 earthquakes reveal normal faulting mechanism with strike-slip component having seismic moment of 3.5E16 N m and 4.3E15 N m respectively. The operation of the Egyptian National Seismic Network (ENSN) since the end of 1997 has significantly enhanced the old picture of earthquake activity across northern Egyptian continental margin whereas; the record-ability (annual rate) has changed from 2-events/year to 54-event/year before and after ENSN respectively. The spatial distribution of earthquakes foci indicated that the activity tends to cluster at three zones: Mediterranean Ridge (MR), Nile Cone (NC) and Eratosthenes Seamount (ERS). However, two seismic gaps are reported along Levant Basin (LEV) and Herodotus Basin (HER).

  5. History of transcontinental railroads and coal mining on the Northern Plains to 1920

    SciTech Connect

    Bryans, W.S.

    1987-01-01

    This history examines the symbiotic relationship between three transcontinental railroads-the Union Pacific, Northern Pacific, and Great Northern-and coal mining in Montana, North Dakota, and Wyoming through 1920. Throughout their dual existence, American railroads and the coal industry enjoyed a mutually beneficial association. On the Northern Plains, however, this partnership assumed new dimensions. There, the coal and rails exerted unique influences upon one another. The location of deposits determined many of the transcontinentals' early decisions, especially route selection. The native fuel also was used to promote settlement on railroad lands. Two of the roads, the Union Pacific and Northern Pacific, held land grants containing valuable deposits. The Great Northern, having no such subsidy, acquired coal lands in northern Montana. On these properties, the three railroads pioneered the region's commercial coal mining industry. Eventually, each formed subsidiaries to direct their coal operations. While much of their production supplied steam locomotives, some was sold to the public. Furthermore, the policies of the Northern Pacific and Great Northern especially enabled their coal to stimulate non-railroad enterprises. In addition, all three provided the transportation which made exploitation by others economically feasible.

  6. The AMeX method: a multipurpose tissue-processing and paraffin-embedding method. II. Extraction of spooled DNA and its application to Southern blot hybridization analysis.

    PubMed Central

    Sato, Y.; Mukai, K.; Matsuno, Y.; Furuya, S.; Kagami, Y.; Miwa, M.; Shimosato, Y.

    1990-01-01

    In our previous report, we described a new fixation and paraffin-embedding method (the AMeX method) that preserves many of the antigens that are normally destroyed by routine formalin fixation. The current study was conducted to examine the preservation of high-molecular-weight DNA in tissues processed by this method. DNA was extracted from AMeX-processed tissue sections after deparaffinization by the same method as that used to extract DNA from fresh tissues. The total amounts of DNA extracted from 10 mg each in wet weight of AMeX-processed and fresh mouse liver tissues were identical. In tissues of malignant lymphoma, the total amount of spooled DNA extracted from 50 sections, each 20 microns thick, was about 8 micrograms/mm2. The electrophoretic pattern of DNA digested with restriction endonucleases on agarose gel from AMeX-processed tissue sections did not differ from that of fresh materials. Southern blot hybridization analysis also revealed that the mobility of specific DNA fragments was identical for AMeX-processed and fresh tissues. The AMeX method was thus proved to be a versatile multipurpose tissue-processing procedure, which is expected to provide important information regarding the correlation between morphology, phenotypic expression, and gene alteration. Images Figure 3 Figure 4 Figure 5A Figure 5B PMID:2407122

  7. Inverse PCR and Quantitative PCR as Alternative Methods to Southern Blotting Analysis to Assess Transgene Copy Number and Characterize the Integration Site in Transgenic Woody Plants.

    PubMed

    Stefano, Biricolti; Patrizia, Bogani; Matteo, Cerboneschi; Massimo, Gori

    2016-06-01

    One of the major unanswered questions with respect to the commercial use of genetic transformation in woody plants is the stability of the transgene expression over several decades within the same individual. Gene expression is strongly affected by the copy number which has been integrated into the plant genome and by the local DNA features close to the integration sites. Because woody plants cannot be subjected to selfing or backcrossing to modify the transgenic allelic structure without affecting the valuable traits of the cultivar, molecular characterization of the transformation event is therefore crucial. After assessing the transgene copy number of a set of apple transgenic clones with Southern blotting, we describe two alternative methods: the first is based on inverse PCR (i-PCR) and the second on the quantitative PCR (q-PCR). The methods produced comparable results with the exception of the data regarding a high copy number clone, but while the q-PCR-based system is rapid and easily adaptable to high throughput systems, the i-PCR-based method can provide information regarding the transformation event and the characteristics of the sequences flanking the transgenic construct. PMID:26895172

  8. IgG western blot for confirmatory diagnosis of equivocal cases of toxoplasmosis by EIA-IgG and fluorescent antibody test.

    PubMed

    Khammari, Imen; Saghrouni, Fatma; Yaacoub, Alia; Gaied Meksi, Sondoss; Ach, Hinda; Garma, Lamia; Fathallah, Akila; Ben Saïd, Moncef

    2013-08-01

    The performance values of available techniques used in serodiagnosis of toxoplasmosis are satisfactory but they raise problems of equivocal and discordant results for very low IgG titers. Recently marketed, LDBio-Toxo II IgG Western blot (IB) showed an excellent correlation with the dye test. We estimated the proportion of equivocal and discordant results between the enzyme immunoassay Platelia Toxo IgG (EIA-IgG) and fluorescent antibody test (FAT) and assessed the usefulness of the IB as a confirmatory test. Out of 2,136 sera collected from pregnant women, 1,644 (77.0%) tested unequivocally positive and 407 (19.0%) were negative in both EIA-IgG and FAT. The remaining 85 (4%) sera showed equivocal or discordant results. Among them, 73 (85.9%) were positive and 12 (14.1%) were negative in IB. Forty-one (89.1%) equivocal sera in EIA-IgG and 46 (86.8%) equivocal sera in FAT were positive in IB. Reducing the cut-off values of both screening techniques improved significantly their sensitivity in detecting very low IgG titers at the expense of their specificity. In conclusion, equivocal results in routine-used techniques and their discordance in determination of the immune status in pregnancy women were not uncommon. IB test appeard to be highly useful in these situations as a confirmatory technique. PMID:24039295

  9. IgG Western Blot for Confirmatory Diagnosis of Equivocal Cases of Toxoplasmosis by EIA-IgG and Fluorescent Antibody Test

    PubMed Central

    Saghrouni, Fatma; Yaacoub, Alia; Gaied Meksi, Sondoss; Ach, Hinda; Garma, Lamia; Fathallah, Akila; Ben Saïd, Moncef

    2013-01-01

    The performance values of available techniques used in serodiagnosis of toxoplasmosis are satisfactory but they raise problems of equivocal and discordant results for very low IgG titers. Recently marketed, LDBio-Toxo II IgG Western blot (IB) showed an excellent correlation with the dye test. We estimated the proportion of equivocal and discordant results between the enzyme immunoassay Platelia Toxo IgG (EIA-IgG) and fluorescent antibody test (FAT) and assessed the usefulness of the IB as a confirmatory test. Out of 2,136 sera collected from pregnant women, 1,644 (77.0%) tested unequivocally positive and 407 (19.0%) were negative in both EIA-IgG and FAT. The remaining 85 (4%) sera showed equivocal or discordant results. Among them, 73 (85.9%) were positive and 12 (14.1%) were negative in IB. Forty-one (89.1%) equivocal sera in EIA-IgG and 46 (86.8%) equivocal sera in FAT were positive in IB. Reducing the cut-off values of both screening techniques improved significantly their sensitivity in detecting very low IgG titers at the expense of their specificity. In conclusion, equivocal results in routine-used techniques and their discordance in determination of the immune status in pregnancy women were not uncommon. IB test appeard to be highly useful in these situations as a confirmatory technique. PMID:24039295

  10. Geologic map of the northern plains of Mars

    USGS Publications Warehouse

    Tanaka, Kenneth L.; Skinner, James A.; Hare, Trent M.

    2005-01-01

    The northern plains of Mars cover nearly a third of the planet and constitute the planet's broadest region of lowlands. Apparently formed early in Mars' history, the northern lowlands served as a repository both for sediments shed from the adjacent ancient highlands and for volcanic flows and deposits from sources within and near the lowlands. Geomorphic evidence for extensive tectonic deformation and reworking of surface materials through release of volatiles occurs throughout the northern plains. In the polar region, Planum Boreum contains evidence for the accumulation of ice and dust, and surrounding dune fields suggest widespread aeolian transport and erosion. The most recent regional- and global-scale maps describing the geology of the northern plains are largely based on Viking Orbiter image data (Dial, 1984; Witbeck and Underwood, 1984; Scott and Tanaka, 1986; Greeley and Guest, 1987; Tanaka and Scott, 1987; Tanaka and others, 1992a; Rotto and Tanaka, 1995; Crumpler and others, 2001; McGill, 2002). These maps reveal highland, plains, volcanic, and polar units based on morphologic character, albedo, and relative ages using local stratigraphic relations and crater counts. This geologic map of the northern plains is the first published map that covers a significant part of Mars using topography and image data from both the Mars Global Surveyor and Mars Odyssey missions. The new data provide a fresh perspective on the geology of the region that reveals many previously unrecognizable units, features, and temporal relations. In addition, we adapted and instituted terrestrial mapping methods and stratigraphic conventions that we think result in a clearer and more objective map. We focus on mapping with the intent of reconstructing the history of geologic activity within the northern plains, including deposition, volcanism, erosion, tectonism, impact cratering, and other processes with the aid of comprehensive crater-density determinations. Mapped areas include all

  11. Cortinarius sect. Armillati in northern Europe.

    PubMed

    Niskanen, Tuula; Kytövuori, Ilkka; Liimatainen, Kare

    2011-01-01

    Cortinarius sect. Armillati (subgenus Telamonia) was studied extensively based on morphology and molecular data. A total of about 1000 specimens, mostly from Fennoscandia, were revised. The nomenclature of the species was confirmed by sequencing the type material. Phylogenetic relationships were inferred by analyses of ITS, and the results were compared with the morphological and ecological data. Based on macro- and micromorphological characters, as well as molecular data, section Armillati contains only the medium to large species with slightly hygrophanous pileus and ± reddish or in some species yellowish brown to rose brown universal veils. The other red-brown-veiled species, previously included in Armillati, seem to belong to at least seven different sections or clades: sect. Anthracini, sect. Boulderenses, sect. Brunneotincti p.p., sect. Cinnabarini, sect. Fulvescentes, /Fuscoperonatus, and /Praestigiosus. Our study recognized six Armillati species from northern Europe: C. armillatus, C. luteo-ornatus, C. paragaudis, and three species described as new, C. pinigaudis, C. roseoarmillatus, and C. suboenochelis. The former three also occur in North America. Two additional species, C. subarmillatus (Japan) and C. quercoarmillatus (Costa Rica), are known outside the area. Based on the phylogenetic analysis, the species associated with deciduous trees, C. armillatus, C. quercoarmillatus, and C. roseoarmillatus, all with dextrinoid, thick-walled spores, formed a separate group from the mainly conifer-associated species, C. luteo-ornatus, C. paragaudis, C. pinigaudis and C. suboenochelis, all with fairly thin to moderately thick-walled, indextrinoid to moderately dextrinoid spores. Descriptions of the northern European species are provided, the distribution is mapped and their taxonomy, ecology, distribution, and relationships are discussed. A total of 64 new sequences of 12 species are reported including 17 sequences from type material. Our study also suggests that

  12. Low Proviral Load is Associated with Indeterminate Western Blot Patterns in Human T-Cell Lymphotropic Virus Type 1 Infected Individuals: Could Punctual Mutations be Related?

    PubMed Central

    Cánepa, Camila; Salido, Jimena; Ruggieri, Matías; Fraile, Sindy; Pataccini, Gabriela; Berini, Carolina; Biglione, Mirna

    2015-01-01

    Background: indeterminate Western blot (WB) patterns are a major concern for diagnosis of human T-cell lymphotropic virus type 1 (HTLV-1) infection, even in non-endemic areas. Objectives: (a) to define the prevalence of indeterminate WB among different populations from Argentina; (b) to evaluate if low proviral load (PVL) is associated with indeterminate WB profiles; and (c) to describe mutations in LTR and tax sequence of these cases. Results: Among 2031 samples, 294 were reactive by screening. Of them, 48 (16.3%) were WB indeterminate and of those 15 (31.3%) were PCR+. Quantitative real-time PCR (qPCR) was performed to 52 HTLV-1+ samples, classified as Group 1 (G1): 25 WB+ samples from individuals with pathologies; Group 2 (G2): 18 WB+ samples from asymptomatic carriers (AC); and Group 3 (G3): 9 seroindeterminate samples from AC. Median PVL was 4.78, 2.38, and 0.15 HTLV-1 copies/100 PBMCs, respectively; a significant difference (p=0.003) was observed. Age and sex were associated with PVL in G1 and G2, respectively. Mutations in the distal and central regions of Tax Responsive Elements (TRE) 1 and 2 of G3 were observed, though not associated with PVL.The 8403A>G mutation of the distal region, previously related to high PVL, was absent in G3 but present in 50% of WB+ samples (p = 0.03). Conclusions: indeterminate WB results confirmed later as HTLV-1 positive may be associated with low PVL levels. Mutations in LTR and tax are described;  their functional relevance remains to be determined. PMID:26516904

  13. Evaluation of the Punch-it™ NA-Sample kit for detecting microbial DNA in blood culture bottles using PCR-reverse blot hybridization assay.

    PubMed

    Kim, Jungho; Wang, Hye-Young; Kim, Seoyong; Park, Soon Deok; Yu, Kwangmin; Kim, Hyo Youl; Uh, Young; Lee, Hyeyoung

    2016-09-01

    DNA extraction efficiency affects the success of PCR-based method applications. The Punch-it™ NA-Sample kit for extracting DNA by using paper chromatography is technically easy to use and requires just two reagents and only 10min to complete. The Punch-it™ NA-Sample kit could be offered as a rapid, accurate, and convenient method for extracting bacterial and fungal DNA from blood culture bottles. We compared the efficiencies of the commercial kit (Punch-it™ NA-Sample kit) and an in-house conventional boiling method with Chelex-100 resin for DNA extraction from blood culture bottles. The efficiency of the two DNA extraction methods was assessed by PCR-reverse blot hybridization assay (PCR-REBA, REBA Sepsis-ID) for detecting Gram positive (GP) bacteria, Gram negative (GN) bacteria, and Candida species with 196 positive and 200 negative blood culture bottles. The detection limits of the two DNA extraction methods were 10(3)CFU/mL for GP bacteria, 10(3)CFU/mL for GN bacteria, and 10(4)CFU/mL for Candida. The sensitivity and specificity of the Punch-it™ NA-Sample kit by REBA Sepsis-ID were 95.4% (187/196) and 100% (200/200), respectively. The overall agreement of the two DNA extraction methods was 98.9% (392/396). Three of four samples showing discrepant results between the two extraction methods were more accurately matched up with the Punch-it™ NA-Sample kit based on conventional culture methods. The results indicated that the Punch-it™ NA-Sample kit extracted bacterial and fungal DNA in blood culture bottles and allowed extracted DNA to be used in molecular assay. PMID:27263831

  14. Identification of reference proteins for Western blot analyses in mouse model systems of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) toxicity.

    PubMed

    Prokopec, Stephenie D; Watson, John D; Pohjanvirta, Raimo; Boutros, Paul C

    2014-01-01

    Western blotting is a well-established, inexpensive and accurate way of measuring protein content. Because of technical variation between wells, normalization is required for valid interpretation of results across multiple samples. Typically this involves the use of one or more endogenous controls to adjust the measured levels of experimental molecules. Although some endogenous controls are widely used, validation is required for each experimental system. This is critical when studying transcriptional-modulators, such as toxicants like 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD).To address this issue, we examined hepatic tissue from 192 mice representing 47 unique combinations of strain, sex, Ahr-genotype, TCDD dose and treatment time. We examined 7 candidate reference proteins in each animal and assessed consistency of protein abundance through: 1) TCDD-induced fold-difference in protein content from basal levels, 2) inter- and intra- animal stability, and 3) the ability of each candidate to reduce instability of the other candidates. Univariate analyses identified HPRT as the most stable protein. Multivariate analysis indicated that stability generally increased with the number of proteins used, but gains from using >3 proteins were small. Lastly, by comparing these new data to our previous studies of mRNA controls on the same animals, we were able to show that the ideal mRNA and protein control-genes are distinct, and use of only 2-3 proteins provides strong stability, unlike in mRNA studies in the same cohort, where larger control-gene batteries were needed. PMID:25329058

  15. Dogs vaccinated with gentamicin-attenuated Leishmania infantum or infected with wild-type parasite can be distinguished by Western blotting.

    PubMed

    Daneshvar, H; Mahmmodi, Z; Kamiabi, H; Phillips, R S; Burchmore, R

    2014-05-01

    An attenuated line of Leishmania infantum (the H-line), developed through exposure to gentamicin, has been shown to protect dogs against canine visceral leishmaniasis. A specific diagnostic test to differentiate dogs vaccinated with the attenuated line from dogs infected with L. infantum wild-type (L. infantum WT) could be a valuable tool in evaluating the effectiveness of canine vaccination. In this study, 28 healthy dogs were allocated into four groups. In Group I and Group II (eight dogs per group), dogs were immunized subcutaneously (s.c.) with L. infantum H-line, and the dogs of Group II challenged s.c. with L. infantum WT, at 2 months post-immunization. In Group III, eight animals were challenged s.c. with L. infantum WT, and four dogs of Group IV were injected s.c. with PBS. We found that sera from vaccinated dogs recognize a 21 kDa antigen of promastigotes of L. infantum H-line but not of L. infantum WT, whereas sera from unvaccinated dogs challenged with L. infantum WT, recognized a 21 kDa antigen of promastigotes of L. infantum WT but not of L. infantum H-line. Sera from dogs challenged with L. infantum WT with prior vaccination with L. infantum H-line, recognized a 21 kDa antigen of both L. infantum WT and L. infantum H-line. These results suggest that the Western blot analysis of antibodies against 21 kDa antigens of L. infantum H-line and WT may be a useful technique for distinguishing between dogs vaccinated with L. infantum H-line and dogs naturally infected with L. infantum WT. PMID:24611833

  16. Evaluation of a New Dot Blot Enzyme Immunoassay (Directigen Flu A+B) for Simultaneous and Differential Detection of Influenza A and B Virus Antigens from Respiratory Samples

    PubMed Central

    Reina, Jordi; Padilla, Emma; Alonso, Fermin; Ruiz de Gopegui, Enrique; Munar, Maria; Mari, Margarita

    2002-01-01

    We report a prospective evaluation of a new dot blot enzyme immunoassay (EIA) method for the direct, rapid, qualitative, simultaneous, and differential detection of the influenza A (IA) and B (IB) virus antigen in different respiratory samples. The EIA method was compared with the shell vial culture system (MDCK cell line) used with the same samples. We studied 160 samples from 93 (58.1%) pediatric patients (hospital emergency room) and from 67 (41.9%) adult patients (sentinel network). Seventy-four(46.2%) samples were considered positive; of them, 46 (62.2%) were from pediatric patients and 28 (37.8%) were from an adult group (P < 0.05), with overall positive values of 49.9% and 41.7%, respectively. All 74 (100%) of the positive samples were isolated in cell culture versus the 68.9% that were detected as positive by the new EIA method (P < 0.05). Of the 41 samples positive for the IA virus, the EIA detected 34 (82.9%) positive samples; of the 33 samples positive for the IB virus, the EIA detected 17 (51.5%) positive samples (P < 0.05). No false-positive reaction was detected with the EIA method (specificity and positive predictive value of 100%). The overall results obtained in the comparison between the new EIA and the shell vial culture had a sensibility of 82.9% and predictive negative values of 92.4% for the IA virus and 51.5% and 84.3%, respectively, for the IB virus. This evaluation shows sensitivity and specificity percentages for the new EIA method that is acceptable for routine use in IA virus detection. The results obtained were worse for IB virus detection, but this new EIA method is actually the only one with the capacity to differentiate between the two influenza viruses. PMID:12202608

  17. Identification of Reference Proteins for Western Blot Analyses in Mouse Model Systems of 2,3,7,8-Tetrachlorodibenzo-P-Dioxin (TCDD) Toxicity

    PubMed Central

    Prokopec, Stephenie D.; Watson, John D.; Pohjanvirta, Raimo; Boutros, Paul C.

    2014-01-01

    Western blotting is a well-established, inexpensive and accurate way of measuring protein content. Because of technical variation between wells, normalization is required for valid interpretation of results across multiple samples. Typically this involves the use of one or more endogenous controls to adjust the measured levels of experimental molecules. Although some endogenous controls are widely used, validation is required for each experimental system. This is critical when studying transcriptional-modulators, such as toxicants like 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD).To address this issue, we examined hepatic tissue from 192 mice representing 47 unique combinations of strain, sex, Ahr-genotype, TCDD dose and treatment time. We examined 7 candidate reference proteins in each animal and assessed consistency of protein abundance through: 1) TCDD-induced fold-difference in protein content from basal levels, 2) inter- and intra- animal stability, and 3) the ability of each candidate to reduce instability of the other candidates. Univariate analyses identified HPRT as the most stable protein. Multivariate analysis indicated that stability generally increased with the number of proteins used, but gains from using >3 proteins were small. Lastly, by comparing these new data to our previous studies of mRNA controls on the same animals, we were able to show that the ideal mRNA and protein control-genes are distinct, and use of only 2–3 proteins provides strong stability, unlike in mRNA studies in the same cohort, where larger control-gene batteries were needed. PMID:25329058

  18. Geothermal systems of northern Nevada

    USGS Publications Warehouse

    Hose, Richard Kenneth; Taylor, Bruce Edward

    1974-01-01

    Hot springs are numerous and nearly uniformly distributed in northern Nevada. Most occur on the flanks of basins, along Basin and Range (late Miocene to Holocene) faults, while some occur in the inner parts of the basins. Surface temperatures of the springs range from slightly above ambient to, boiling; some springs are superheated. Maximum subsurface water temperatures calculated on the basis of quartz solubility range as high as 252?C, although most are below 190?C. Flows range from a trickle to several hundred liters per minute. The Nevada geothermal systems differ markedly from the power-producing system at The Geysers, Calif., and from those areas with a high potential, for power production (e.g., Yellowstone Park, Wyo.; Jemez Mountains, N. Mex.). These other systems are associated with Quaternary felsic volcanic rocks and probably derive their heat from cooling magma rather high in the crust. In northern Nevada, however, felsic volcanic rocks are virtually all older than 10 million years, and. analogous magmatic heat sources are, therefore, probably lacking. Nevada is part of an area of much higher average heat flow than the rest of the United States. In north-central Nevada, geothermal gradients are as great as 64?C per kilometer in bedrock and even higher in basin fill. The high gradients probably result from a combination of thin crust and high temperature upper mantle. We suggest that the geothermal systems of northern Nevada result from circulation of meteoric waters along Basin and Range faults and that their temperature chiefly depends upon (1) depth of circulation and (2) the geothermal gradient near the faults.

  19. Crustal structure of northern California

    NASA Astrophysics Data System (ADS)

    Godfrey, Nicola Jane

    This thesis presents work from two regions in northern California, using seismic data collected during the Mendocino Triple Junction Experiment (1993 and 1994), and USGS data collected in 1977. Much of California geology records subduction processes active during much of the Mesozoic. About 29 Ma ago, the East Pacific Rise began interacting with the California subduction margin, and two triple junctions formed. One moved north and one moved south, with the San Andreas transform system between them. The northern triple junction, the Mendocino triple junction, is currently situated close to Cape Mendocino, northern California. Northern California geology now has the added complication of processes associated with the northward moving triple junction and lengthening San Andreas fault system. Chapters 1 and 2 focus on the Great Valley, a forearc basin associated with Mesozoic subduction. The three major parts of the subduction system are the magmatic arc (Sierra Nevada batholith), forearc basin (Great Valley basin) and accretionary prism (Franciscan complex). Chapter 1 presents evidence from seismic data, for a complete ophiolitic sequence, including an unserpentinized mantle section (velocities of 8.1 km.ssp{-1} at 5-18 km depth) beneath the northernmost Great Valley. A combination velocity/density model shows the ophiolite is underlain by low-density material associated with the Sierra Nevada. Chapter 2 presents geophysical models, (seismic reflection, refraction, gravity and magnetic models) published by different authors over the last 14 years from the entire Great Valley, to explore the nature of the Great Valley ophiolite along the length of the forearc basin. Chapter 3 focuses offshore, west of the San Andreas fault, on an anomalous piece of crust, the Vizcaino block, situated immediately southwest of the Mendocino triple junction. This chapter discusses the crustal thickness of the Vizcaino block, the nature of its accretionary prism basement, and deformation

  20. Geothermal development plan: northern Arizona

    SciTech Connect

    White, D.H.; Goldstone, L.A.

    1981-01-01

    Much of the northern counties (Apache, Coconino, Gila, Mohave, Navajo and Yavapai) is located in the Colorado Plateau province, a region of low geothermal potential. Two areas that do show some potential are the Flagstaff - San Francisco Peaks area and the Springerville area. Flagstaff is rapidly becoming the manufacturing center of Arizona and will have many opportunities to use geothermal energy to satisfy part of its increasing need for energy. Using a computer simulation model, projections of geothermal energy on line as a function of time are made for both private and city-owned utility development of a resource.

  1. General Practice in Northern Norway

    PubMed Central

    Black, D. P.

    1975-01-01

    A study was made of general practice in northern Norway where conditions are similar to parts of rural Canada. The Norwegian general practitioner has developed expertise in the preventive and psychosocial aspects of practice and the team concept is highly developed. Since the general practitioner is separated from the hospital, his facilities for procedures and diagnostic workups are primitive. Involvement of general practitioners in medical education is not yet well developed although all new graduates spend a compulsory period in rural practice. PMID:20469187

  2. Clinical performance of the Multispot HIV-1/HIV-2 rapid test to correctly differentiate HIV-2 from HIV-1 infection in screening algorithms using third and fourth generation assays and to identify cross reactivity with the HIV-1 Western Blot

    PubMed Central

    Ramos, Eric M.; Harb, Socorro; Dragavon, Joan; Coombs, Robert W.

    2014-01-01

    Background An accurate and rapid serologic method to differentiate HIV-2 from HIV-1 infection is required since the confirmatory HIV-1 Western Blot (WB) may demonstrate cross-reactivity with HIV-2 antibodies. Objectives To evaluate the performance of the Bio-Rad Multispot HIV-1/HIV-2 rapid assay as a supplemental test to correctly identify HIV-2 infection and identify HIV-1 WB cross-reactivity with HIV-2 in clinical samples tested at an academic medical center. Study design Between August 2008 and July 2012, clinical samples were screened for HIV using either 3rd-or 4th-generation HIV-1/2 antibody or combination antibody and HIV-1 p24 antigen assays, respectively. All repeatedly reactive samples were reflexed for Multispot rapid testing. Multispot HIV-2 and HIV-1 and HIV-2-reactive samples were further tested using an HIV-2 immunoblot assay and HIV-1 or HIV-2 RNA assays when possible. The HIV-1 WB was performed routinely for additional confirmation and to assess for HIV-2 antibody cross-reactivity. Results Of 46,061 samples screened, 890 (89.6%) of 993 repeatedly reactive samples were also Multispot-reactive: 882 for HIV-1; three for only HIV-2; and five for both HIV-1 and HIV-2. All three HIV-2-only Multispot-positives along with a single dually reactive HIV-1/2 Multispot-positive were also HIV-2 immunoblot-positive; the latter was HIV-1 RNA negative and HIV-2 RNA positive. Conclusions The Multispot rapid test performed well as a supplemental test for HIV-1/2 diagnostic testing. Four new HIV-2 infections (0.45%) were identified from among 890 Multispot-reactive tests. The use of HIV-1 WB alone to confirm HIV-1/2 screening assays may underestimate the true prevalence of HIV-2 infection in the United States. PMID:24342468

  3. Response of sugar maple to calcium addition to northern hardwood forest.

    PubMed

    Juice, Stephanie M; Fahey, Timothy J; Siccama, Thomas G; Driscoll, Charles T; Denny, Ellen G; Eagar, Christopher; Cleavitt, Natalie L; Minocha, Rakesh; Richardson, Andrew D

    2006-05-01

    Watershed budget studies at the Hubbard Brook Experimental Forest (HBEF), New Hampshire, USA, have demonstrated high calcium depletion of soil during the 20th century due, in part, to acid deposition. Over the past 25 years, tree growth (especially for sugar maple) has declined on the experimental watersheds at the HBEF. In October 1999, 0.85 Mg Ca/ha was added to Watershed 1 (W1) at the HBEF in the form of wollastonite (CaSiO3), a treatment that, by summer 2002, had raised the pH in the Oie horizon from 3.8 to 5.0 and, in the Oa horizon, from 3.9 to 4.2. We measured the response of sugar maple to the calcium fertilization treatment on W1. Foliar calcium concentration of canopy sugar maples in W1 increased markedly beginning the second year after treatment, and foliar manganese declined in years four and five. By 2005, the crown condition of sugar maple was much healthier in the treated watershed as compared with the untreated reference watershed (W6). Following high seed production in 2000 and 2002, the density of sugar maple seedlings increased significantly on W1 in comparison with W6 in 2001 and 2003. Survivorship of the 2003 cohort through July 2005 was much higher on W1 (36.6%) than W6 (10.2%). In 2003, sugar maple germinants on W1 were approximately 50% larger than those in reference plots, and foliar chlorophyll concentrations were significantly greater (0.27 g/m2 vs. 0.23 g/m2 leaf area). Foliage and fine-root calcium concentrations were roughly twice as high, and manganese concentrations twice as low in the treated than the reference seedlings in 2003 and 2004. Mycorrhizal colonization of seedlings was also much greater in the treated (22.4% of root length) than the reference sites (4.4%). A similar, though less dramatic, difference was observed for mycorrhizal colonization of mature sugar maples (56% vs. 35%). These results reinforce and extend other regional observations that sugar maple decline in the northeastern United States and southern Canada is caused in part by anthropogenic effects on soil calcium status, but the causal interactions among inorganic nutrition, physiological stress, mycorrhizal colonization, and seedling growth and health remain to be established. PMID:16761605

  4. Comparative ability of northern pintails, gadwalls, and northern shovelers to metabolize foods

    USGS Publications Warehouse

    Miller, M.R.

    1984-01-01

    Feeding trials were used to compare the ability of northern pintails (Anas acuta), gadwalls (A. strepera), and northern shovelers (A. clypeata) to metabolize energy from a turkey starter ration, alfalfa pellets, and common barnyardgrass (Echinochloa crusgalli) seeds. No differences (P > 0.05) were detected among the three species for any of the three foods (kg body weight/day basis), for dry matter intake (DMI), body weight gain (BWG), apparent metabolizable energy (AME), or metabolized energy (MEE) on any given diet consumed in quantities large enough to promote body weight gain. The AME content of alfalfa was 57% less than the value for turkey starter and 50% less than for barnyardgrass seeds. All three species metabolized more energy and gained weight faster when fed turkey starter. Energy modeling may be facilitated if additional research verifies that all species of dabbling ducks have equal ability to obtain energy from foods available to them in the wild. Behavioral and morphological factors may be more important in defining feeding niches than digestive physiology, at least for the three duck species tested, at the time of year of the experiments, and within the limits of the quality of foods used.

  5. Variability of Carbon Exchanges Between Two Contrasting Northern Peatlands

    NASA Astrophysics Data System (ADS)

    Roulet, N. T.; Nilsson, M.

    2008-12-01

    Northern peatland contain about one quarter of the world's terrestrial carbon. It appears that many peatlands still remains a small, but persistent sinks of carbon dioxide and sources of methane. The sink strength is small compared to actively growing boreal forests but equal to the Holocene average peatland carbon accumulation. This suggests that the function of northern peatlands, with regard to C sequestration, has not change relative to the Holocene average uptake. In contrast to forested ecosystems there have been few long-term continuous measurements of the components of the C balance of peatlands ecosystems. In addition to measurements of net ecosystem exchange and net methane emission (or uptake), the C balance of peatlands requires accurate estimates of the loss of carbon dissolved in runoff. Multi-year measurements of these three major exchanges have been made in contrasting northern peatlands: Mer Bleue, a raised ombrotrophic bog located at the boreal - temperate boundary in eastern Canada, and Degero Stormyr, a mineral poor, oligotrophic fen located in northern Sweden. Despite very different plant communities and moisture regimes the long-term average NEE, methane exchange and net loss of carbon dissolved in water are surprisingly similar in these two systems. However, Mer Bleue has a much greater inter-annual variability in the exchanges than does Degero Stormyr peatland. The difference in exchanges appears related to differences in the variability in moisture supply to the vegetation layer and water storage in the peat. In the early 1990s, the eminent peatland ecologist, Eville Gorham estimated, with few observations, the relative importance of the C balance components of northern peatlands. The multi-year records indicate that these early estimates with reasonable good within an order of magnitude.

  6. Water Ice Albedo Variations on the Martian Northern Polar Cap

    NASA Technical Reports Server (NTRS)

    Hale, A. S.; Bass, D. S.; Tamppari, L. K.

    2003-01-01

    The Viking Orbiters determined that the surface of Mars northern residual cap is water ice. Many researchers have related observed atmospheric water vapor abundances to seasonal exchange between reservoirs such as the polar caps, but the extent to which the exchange between the surface and the atmosphere remains uncertain. Early studies of the ice coverage and albedo of the northern residual Martian polar cap using Mariner 9 and Viking images reported that there were substantial internannual differences in ice deposition on the polar cap, a result which suggested a highly variable Martian climate. However, some of the data used in these studies were obtained at differing values of heliocentric solar longitude (L(sub s)). Reevaluation of this dataset indicated that the residual cap undergoes seasonal brightening throughout the summer, and indicated that this process repeats from year to year. In this study we continue to compare Mariner 9 and Viking Orbiter imaging observations and thermal data of the north residual polar cap to data acquired with Mars Global Surveyor s Mars Orbiter Camera (MOC) instrument. In the current study, our goal is to examine all released data from MGS MOC in the northern summer season, along with applicable TES data in order to better understand the albedo variations in the northern summer and their implications on water transport. To date, work has focused primarily on the MOC dataset. In 1999, data acquisition of the northern polar regions began at L(sub s) = 107, although there was little north polar data acquired from L(sub s)= 107 to L(sub s) = 109. We examined a total of 409 images from L(sub s) = 107 to L(sub s)=148. We have also examined data from 2000 from L(sub s)= 93 to L(sub s)= 110; additional progress is ongoing. Here we present a progress report of our observations, and continue to determine their implications for the Martian water cycle.

  7. Phylogeographic Analysis of Mitochondrial DNA in Northern Asian Populations

    PubMed Central

    Derenko, Miroslava ; Malyarchuk, Boris ; Grzybowski, Tomasz ; Denisova, Galina ; Dambueva, Irina ; Perkova, Maria ; Dorzhu, Choduraa ; Luzina, Faina ; Lee, Hong Kyu ; Vanecek, Tomas ; Villems, Richard ; Zakharov, Ilia 

    2007-01-01

    To elucidate the human colonization process of northern Asia and human dispersals to the Americas, a diverse subset of 71 mitochondrial DNA (mtDNA) lineages was chosen for complete genome sequencing from the collection of 1,432 control-region sequences sampled from 18 autochthonous populations of northern, central, eastern, and southwestern Asia. On the basis of complete mtDNA sequencing, we have revised the classification of haplogroups A, D2, G1, M7, and I; identified six new subhaplogroups (I4, N1e, G1c, M7d, M7e, and J1b2a); and fully characterized haplogroups N1a and G1b, which were previously described only by the first hypervariable segment (HVS1) sequencing and coding-region restriction-fragment–length polymorphism analysis. Our findings indicate that the southern Siberian mtDNA pool harbors several lineages associated with the Late Upper Paleolithic and/or early Neolithic dispersals from both eastern Asia and southwestern Asia/southern Caucasus. Moreover, the phylogeography of the D2 lineages suggests that southern Siberia is likely to be a geographical source for the last postglacial maximum spread of this subhaplogroup to northern Siberia and that the expansion of the D2b branch occurred in Beringia ∼7,000 years ago. In general, a detailed analysis of mtDNA gene pools of northern Asians provides the additional evidence to rule out the existence of a northern Asian route for the initial human colonization of Asia. PMID:17924343

  8. Illuminating Northern California's Active Faults

    NASA Astrophysics Data System (ADS)

    Prentice, Carol S.; Crosby, Christopher J.; Whitehill, Caroline S.; Arrowsmith, J. Ramón; Furlong, Kevin P.; Phillips, David A.

    2009-02-01

    Newly acquired light detection and ranging (lidar) topographic data provide a powerful community resource for the study of landforms associated with the plate boundary faults of northern California (Figure 1). In the spring of 2007, GeoEarthScope, a component of the EarthScope Facility construction project funded by the U.S. National Science Foundation, acquired approximately 2000 square kilometers of airborne lidar topographic data along major active fault zones of northern California. These data are now freely available in point cloud (x, y, z coordinate data for every laser return), digital elevation model (DEM), and KMZ (zipped Keyhole Markup Language, for use in Google Earth™ and other similar software) formats through the GEON OpenTopography Portal (http://www.OpenTopography.org/data). Importantly, vegetation can be digitally removed from lidar data, producing high-resolution images (0.5- or 1.0-meter DEMs) of the ground surface beneath forested regions that reveal landforms typically obscured by vegetation canopy (Figure 2).

  9. Groundwater management in northern Iraq

    NASA Astrophysics Data System (ADS)

    Stevanovic, Zoran; Iurkiewicz, Adrian

    2009-03-01

    Groundwater is vital and the sole resource in most of the studied region of northern Iraq. It has a significant role in agriculture, water supply and health, and the elimination of poverty in rural areas. Although Iraq is currently dramatically disturbed by complex political and socio-economic problems, in its northern part, i.e. the Kurdish-inhabited region, fast urbanization and economic expansion are visible everywhere. Monitoring and water management schemes are necessary to prevent aquifer over-exploitation in the region. Artificial recharge with temporary runoff water, construction of subsurface dams and several other aquifer management and regulation measures have been designed, and some implemented, in order to improve the water situation. Recommendations, presented to the local professionals and decision-makers in water management, include creation of Water Master Plans and Water User Associations, synchronization of drilling programmes, rehabilitation of the existing well fields, opening of new well fields, and the incorporation of new spring intakes in some areas with large groundwater reserves, as well as construction of numerous small-scale schemes for initial in situ water treatment where saline groundwater is present.

  10. Modeling the above and below ground carbon and nitrogen stocks in northern high latitude terrestrial ecosystems

    NASA Astrophysics Data System (ADS)

    ElMasri, B.; Jain, A. K.

    2012-12-01

    Climate change is expected to cause warming in the northern high latitudes, but it is still uncertain what the respond of the northern high latitudes ecosystem will be to such warming. One of the biggest scientific questions is to determine whether northern high latitude ecosystem are or will act as a terrestrial carbon sink or source. Therefore, it is essential to understand and quantify the biogeochemical cycle of the northern high latitude ecosystems in order to predict their respond to climate change. Using a land surface model, the Integrated Science Assessment Model (ISAM) with its coupled carbon-nitrogen cycle, we provide a detail quantification of the carbon and nitrogen in the vegetation pools and the soil carbon for the northern high latitude ecosystems. We focus on soil carbon and vegetation carbon and nitrogen, though we provide results for gross primary production (GPP), autotrophic respiration (Ra), net primary production (NPP), net ecosystem exchange (NEE), and heterotrophic respiration (Rh). In addition, we examine the effect of nitrogen limitation on the carbon fluxes and soil carbon. We present the results for several flux tower sites representative of the tundra and the boreal ecosystems as well as for the northern high latitude region. Our results provide a comprehensive assessment of below and above ground carbon and nitrogen pools in the northern high latitude and the model calibrated parameters can be used to improve the results of other land surface models.

  11. A Right to Be Heard: Learning from Learners with Additional Needs in Literacy

    ERIC Educational Resources Information Center

    McPhillips, Therese; Shevlin, Michael; Long, Louise

    2012-01-01

    This study aimed to explore the consultation experiences of pupils who have additional needs in literacy. An opportunistic sample of eight schools--four in Northern Ireland and four in the Republic of Ireland--were chosen by the researchers; selected pupils were receiving additional literacy support. Focus group discussions and arts-based creative…

  12. Localization of eight additional genes in the human major histocompatibility complex, including the gene encoding the casein kinase II {beta} subunit (CSNK2B)

    SciTech Connect

    Albertella, M.R.; Jones, H.; Thomson, W.

    1996-09-01

    A wide range of autoimmune and other diseases are known to be associated with the major histocompatibility complex. Many of these diseases are linked to the genes encoding the polymorphic histocompatibility complex. Many of these diseases are linked to the genes encoding the polymorphic histocompatibility antigens in the class I and class II regions, but some appear to be more strongly associated with genes in the central 1100-kb class III region, making it important to characterize this region fully for the presence of novel genes. An {approximately}220-kb segment of DNA in the class III region separating the Hsp70 (HSPA1L) and BAT1 (D6S8IE) genes, which was previously known to contain 14 genes. Genomic DNA fragments spanning the gaps between the known genes were used as probes to isolate cDNAs corresponding to five new genes within this region. Evidence from Northern blot analysis and exon trapping experiments that suggested the presence of at least two more new genes was also obtained. Partial cDNA and complete exonic genomic sequencing of one of the new genes has identified it as the casein kinase II{beta} subunit (CSNK2B). Two of the other novel genes lie within a region syntenic to that implicated in susceptibility to experimental allergic orchitis in the mouse, an autoimmune disease of the testis, and represent additional candidates for the Orch-1 locus associated with this disease. In addition, characterization of the 13-kb intergenic gap separating the RD (D6545) and G11 (D6S60E) genes has revealed the presence of a gene encoding a 1246-amino-acid polypeptide that shows significant sequence similarity to the yeast anti-viral Ski2p gene product. 49 refs., 8 figs.

  13. Stratospheric influence on Northern Hemisphere winter climate variability

    NASA Astrophysics Data System (ADS)

    Ouzeau, Gaelle; Douville, Herve; Saint Martin, David

    2010-05-01

    Despite significant improvements in observing and data assimilation systems, long-range dynamical forecasting remains a difficult challenge for the climate modelling community. The skill of operational seasonal forecasting systems is particularly poor in the northern extratropics where seas surface temperature (SST) has a weaker influence than in the Tropics. It is therefore relevant to look for additional potential sources of long-range climate predictability in the stratosphere using ensembles of global atmospheric simulations. Besides a control experiment where the ARPEGE-Climat model is only driven by SST, parallel simulations have been performed in which an additional control on climate variability has been accounted for through the nudging of the northern extratropical stratosphere towards the ERA40 reanalysis. Though idealized, this original experiment design allows us to compare the relative contribution of the lower and upper boundary forcings on the simulated tropospheric variability. Results show that the stratospheric nudging improves the climatology and interannual variability of the mid-latitude troposphere, especially in winter in the Northern Hemisphere. Major impacts are found in particular on the simulation of the Arctic and North Atlantic oscillations (AO and NAO). Case studies were carried out for the 1976-1977 and 1988-1989 winters, corresponding to extreme phases of the AO. Results confirm the robustness of the positive impact of the nudging, especially for winter 1976-1977 corresponding to relatively weak SST anomalies in the tropical Pacific. A sensitivity study to the model resolution shows that a well-resolved stratosphere is not necessary for the nudging to be efficient. Besides seasonal mean results, analysis of the day-to-day variability in winter allowed us to better understand the stratospheric polar vortex influence on the tropospheric circulation in the Northern Hemisphere mid-latitudes.

  14. Nitrogen Inputs via Nitrogen Fixation in Northern Plants and Soils

    NASA Astrophysics Data System (ADS)

    Thorp, N. R.; Wieder, R. K.; Vile, M. A.

    2015-12-01

    Dominated by cold and often acidic water logged environments, mineralization of organic matter is slow in the majority of northern ecosystems. Measures of extractable ammonium and nitrate are generally low and can be undetectable in peat pore waters. Despite this apparent nitrogen limitation, many of these environments produce deep deposits of soil organic matter. Biological nitrogen fixation carried out by autotrophic and heterotrophic diazotrophs associated with cryptograms provides the majority of known nitrogen inputs in these northern ecosystems. Nitrogen fixation was assessed in a variety of northern soils within rhizospheres of dominant plant communities. We investigated the availability of this newly fixed nitrogen to the vascular plant community in nitrogen limited northern plant communities. We tracked nitrogen flow from 15N2 gas fixed in Sphagnum mosses into tissues of two native vascular plant species, boreal cranberry (Vaccinium oxycoccus) and black spruce (Picea mariana). 15N-labeled Sphagnum microcosms were grown within variable mesh size exclusion/inclusion fabrics in a nitrogen addition experiment in situ in order to investigate the role of mycorrhizal fungi in the uptake of newly fixed nitrogen. Up to 24% of daily fixed 15N label was transferred to vascular plant tissues during 2 months. Nitrogen addition resulted in decreased N2 fixation rates; however, with higher nitrogen availability there was a higher rate of 15N label uptake into the vascular plants, likely the result of increased production of dissolved organic nitrogen. Reliance on mycorrhizal networks for nitrogen acquisition was indicated by nitrogen isotope fractionation patterns. Moreover, N2 fixation activities in mosses were stimulated when vascular plants were grown in moss microcosms versus "moss only" treatments. Results indicate that bog vascular plants may derive considerable nitrogen from atmospheric N2 biologically fixed within Sphagnum mosses. This work demonstrates that

  15. The Alcoholism Situation in a Northern City

    ERIC Educational Resources Information Center

    Martynov, M. Iu.; Martynova, D. Iu.

    2012-01-01

    Alcohol abuse in Russia has been increasing in recent years, especially in northern regions, as has the incidence of alcohol-related disease rates. A survey was conducted in Surgut (the Khanty-Mansi autonomous okrug) that determined the factors lending to the prevalence of alcohol abuse among the population of the northern city and assessed the…

  16. Northern Desegregation: A Tale of Two Cities

    ERIC Educational Resources Information Center

    Danns, Dionne

    2011-01-01

    Studies on northern desegregation have focused on political strategies, the role of the courts, the responsibility of the federal government (HEW), and barriers to northern desegregation. Some have conducted individual case studies and comparative studies, and others have examined a number of cities. This article examines the way school…

  17. Northern Parkway PTA Makes Health a Habit

    ERIC Educational Resources Information Center

    Ferdinand, Marilyn

    2011-01-01

    Health and fitness have been on the agenda of Northern Parkway Elementary School for quite some time, thanks to the concerted efforts of its involved and active PTA officers and members. For the past five years, the Northern Parkway PTA has held a popular and well-attended Family Fun and Fitness Night and has complemented the activities and…

  18. Political Strategies in Northern School Desegregation.

    ERIC Educational Resources Information Center

    Kirby, David J.; And Others

    This is a study of school desegregation as it was fought in 91 Northern and Western Cities between 1963 and 1969. In the summer of 1963, the de facto school segregation issue exploded in dozens of Northern cities; in the next 6 years school integration was front page news in most cities. What this book attempts to do in effect is to record a…

  19. Environmental overview of geothermal development: northern Nevada

    SciTech Connect

    Slemmons, D.B.; Stroh, J.M.; Whitney, R.A.

    1980-08-01

    Regional environmental problems and issues associated with geothermal development in northern Nevada are studied to facilitate environmental assessment of potential geothermal resources. The various issues discussed are: environmental geology, seismicity of northern Nevada, hydrology and water quality, air quality, Nevada ecosystems, noise effects, socio-economic impacts, and cultural resources and archeological values. (MHR)

  20. Meeting Northern Arizona's Supported Employment Training Needs.

    ERIC Educational Resources Information Center

    Martin, William E., Jr.; And Others

    In 1989 Northern Arizona University established a Supported Employment Training Center (SETC) to increase the number of trained job coaches in northern Arizona and provide knowledge and skills in supported employment to personnel from cooperating schools and agencies. First-year SETC activities focused on assessment of the training needs of…

  1. Ultraviolet resources over Northern Eurasia.

    PubMed

    Chubarova, Natalia; Zhdanova, Yekaterina

    2013-10-01

    We propose a new climatology of UV resources over Northern Eurasia, which includes the assessments of both detrimental (erythema) and positive (vitamin D synthesis) effects of ultraviolet radiation on human health. The UV resources are defined by using several classes and subclasses - UV deficiency, UV optimum, and UV excess - for 6 different skin types. To better quantifying the vitamin D irradiance threshold we accounted for an open body fraction S as a function of effective air temperature. The spatial and temporal distribution of UV resources was estimated by radiative transfer (RT) modeling (8 stream DISORT RT code) with 1×1° grid and monthly resolution. For this purpose special datasets of main input geophysical parameters (total ozone content, aerosol characteristics, surface UV albedo, UV cloud modification factor) have been created over the territory of Northern Eurasia. The new approaches were used to retrieve aerosol parameters and cloud modification factor in the UV spectral region. As a result, the UV resources were obtained for clear-sky and mean cloudy conditions for different skin types. We show that the distribution of UV deficiency, UV optimum and UV excess is regulated by various geophysical parameters (mainly, total ozone, cloudiness and open body fraction) and can significantly deviate from latitudinal dependence. We also show that the UV optimum conditions can be simultaneously observed for people with different skin types (for example, for 4-5 skin types at the same time in spring over Western Europe). These UV optimum conditions for different skin types occupy a much larger territory over Europe than that over Asia. PMID:23933245

  2. Differential Proteomic Analysis of Noncardia Gastric Cancer from Individuals of Northern Brazil

    PubMed Central

    Leal, Mariana Ferreira; Chung, Janete; Calcagno, Danielle Queiroz; Assumpção, Paulo Pimentel; Demachki, Samia; da Silva, Ismael Dale Cotrim Guerreiro; Chammas, Roger; Burbano, Rommel Rodríguez; de Arruda Cardoso Smith, Marília

    2012-01-01

    Gastric cancer is the second leading cause of cancer-related death worldwide. The identification of new cancer biomarkers is necessary to reduce the mortality rates through the development of new screening assays and early diagnosis, as well as new target therapies. In this study, we performed a proteomic analysis of noncardia gastric neoplasias of individuals from Northern Brazil. The proteins were analyzed by two-dimensional electrophoresis and mass spectrometry. For the identification of differentially expressed proteins, we used statistical tests with bootstrapping resampling to control the type I error in the multiple comparison analyses. We identified 111 proteins involved in gastric carcinogenesis. The computational analysis revealed several proteins involved in the energy production processes and reinforced the Warburg effect in gastric cancer. ENO1 and HSPB1 expression were further evaluated. ENO1 was selected due to its role in aerobic glycolysis that may contribute to the Warburg effect. Although we observed two up-regulated spots of ENO1 in the proteomic analysis, the mean expression of ENO1 was reduced in gastric tumors by western blot. However, mean ENO1 expression seems to increase in more invasive tumors. This lack of correlation between proteomic and western blot analyses may be due to the presence of other ENO1 spots that present a slightly reduced expression, but with a high impact in the mean protein expression. In neoplasias, HSPB1 is induced by cellular stress to protect cells against apoptosis. In the present study, HSPB1 presented an elevated protein and mRNA expression in a subset of gastric cancer samples. However, no association was observed between HSPB1 expression and clinicopathological characteristics. Here, we identified several possible biomarkers of gastric cancer in individuals from Northern Brazil. These biomarkers may be useful for the assessment of prognosis and stratification for therapy if validated in larger clinical study

  3. Europa's Northern Trailing Hemisphere: Lineament Stratigraphic Framework

    NASA Technical Reports Server (NTRS)

    Figueredo, P. H.; Hare, T.; Ricq, E.; Strom, K.; Greeley, R.; Tanaka, K.; Senske, D.

    2004-01-01

    Knowledge of the global distribution of Europan geologic units in time and space is a necessary step for the synthesis of the results of the Galileo mission and in preparation for future exploration (namely, by JIMO) of the satellite. We have initiated the production of the first Global Geological Map of Europa. As a base map, we use the recently published global photomosaic of Europa (U.S.G.S. Map I-2757) and additional Galileo SSI images at their original resolution. The map is being produced entirely on GIS format for analysis and combination with other datasets [1]. One of the main objectives of this project is to establish a global stratigraphic framework for Europa. In the absence of a well-developed cratering record, this goal will be achieved using the satellite s global network of lineaments (ridges, ridge complexes and bands; cf. [2]). Here we present the preliminary stratigraphic framework synthesized from the sequence of lineaments derived for the northern trailing hemisphere of Europa (Figure 1, below), and we discuss its significance and some emerging implications.

  4. Middle Cretaceous dinosaur assemblages from northern Brazil and northern Africa and their implications for northern Gondwanan composition

    NASA Astrophysics Data System (ADS)

    Candeiro, Carlos Roberto A.

    2015-08-01

    Dinosaurs are one of the most dominant groups in Cretaceous reptilian faunas. A summary of their record in northern Brazil and northern Africa during the middle of the Cretaceous Period (Aptian-Cenomanian) is presented here. Dinosaurs are represented by 32 species (three ornithischians, six sauropods and 23 theropods) from Brazil, Egypt, Lybia, Morocco, Niger, Sudan and Tunisia. These dinosaur assemblages provide fundamental data about distribution and composition of sauropods and theropods in northern Gondwana during the middle of the Cretaceous Period and confirm these assemblages to be among the most important dinosaur faunas in the north Gondwana areas.

  5. Oak ecosystem succession of the Northern Caucasus

    NASA Astrophysics Data System (ADS)

    Chalaya, Elena; Slepykh, Victor; Efimenko, Natalia; Povolotskaya, Nina

    2014-05-01

    English oak (Quercus robur L.) along with its well-known good properties has a high sanitary-hygienic and curative potential. Its volatile metabolites (VM) influence bacteriostatically Staphylococcus aureus 209r, oppressing it in vitro by 85% compared with the control, and Escherichia coli by 45%. There is the least amount of epiphytic microorganisms on the leaves of Q. robur L. compared with some tree species [1]. In addition, VM of Q. robur L. have direct milieu (hypotensive) effects on the organism under its canopy, lowering blood pressure by 20-25 mm Hg [2]. A.P. Kazankin (1993) [4] calculated the prehistoric formula of forest species composition of Caucasian Mineral Waters region (Northern Caucasus): 6Qr3Crp1Fr which was based on the theory of calcium-magnesium absorption complex. According to the theory, underground mineral water, soil, forest litter and the leaves of ground vegetation of the area have the same ratio of the cations Ca and Mg - calcium-magnesium index [3]. Hence oldgrowth in the region consisted of oak (Qr) by 60%, hornbeam (Carpinus-Crp) by 30% and ash (Fraxinus-Fr) only by 10%. Currently, the formula of the forests of the region has been changed by man: 5Fr3Crp2Qr. The proportion of oak forests has decreased to 20%, the proportion of ash has increased by 50%, but the proportion of hornbeam hasn't changed. So it is relevant to restore oak forests of the region in the former ratio to other forest-forming species - ash and hornbeam. Taking into consideration the change of economic formation of society in Russia, it is extremely important to restore natural seed oak forests. Therein the luminance of surface areal is a limiting factor. We have calculated that the natural recovery of oak forests is possible providing observation of optimal moisture standards and soil fertility in combination with solar light within 10-24% from the light of open space. Measures for promotion of oak natural regeneration in mountain resorts of the Northern Caucasus

  6. Demographic response of northern spotted owls to barred owl removal

    USGS Publications Warehouse

    Diller, V. Lowell; Hamm, Keith A; Early, Desiree A; Lamphear, David W; Katie Dugger; Yackulic, Charles B.; Schwarz, Carl J.; Carlson, Peter C.; McDonald, Trent L.

    2016-01-01

    the treated areas and 0.822 for the untreated areas. Derived estimates of population change on both study areas showed the same general decline before removal with an estimated slope of –0.0036 per year. Following removal, the rate of population change on the treated areas increased to an average of 1.029 but decreased to an average of 0.870 on the untreated areas. The results from this first experiment demonstrated that lethal removal of barred owls allowed the recovery of northern spotted owl populations in the treated portions of our study area. If additional federally funded barred owl removal experiments provide similar results, this could be the foundation for development of a long-term conservation strategy for northern spotted owls.

  7. Incorporation of additives into polymers

    DOEpatents

    McCleskey, T. Mark; Yates, Matthew Z.

    2003-07-29

    There has been invented a method for incorporating additives into polymers comprising: (a) forming an aqueous or alcohol-based colloidal system of the polymer; (b) emulsifying the colloidal system with a compressed fluid; and (c) contacting the colloidal polymer with the additive in the presence of the compressed fluid. The colloidal polymer can be contacted with the additive by having the additive in the compressed fluid used for emulsification or by adding the additive to the colloidal system before or after emulsification with the compressed fluid. The invention process can be carried out either as a batch process or as a continuous on-line process.

  8. Deciphering the roles of multiple additives in organocatalyzed Michael additions.

    PubMed

    Günler, Z Inci; Companyó, Xavier; Alfonso, Ignacio; Burés, Jordi; Jimeno, Ciril; Pericàs, Miquel A

    2016-05-21

    The synergistic effects of multiple additives (water and acetic acid) on the asymmetric Michael addition of acetone to nitrostyrene catalyzed by primary amine-thioureas (PAT) were precisely determined. Acetic acid facilitates hydrolysis of the imine intermediates, thus leading to catalytic behavior, and minimizes the formation of the double addition side product. In contrast, water slows down the reaction but minimizes catalyst deactivation, eventually leading to higher final yields. PMID:27128165

  9. The puzzling origin of the Martian Northern Lowlands.

    NASA Astrophysics Data System (ADS)

    Altieri, F.; Carrozzo, G.; Carli, C.; Geminale, A.; Bellucci, G.

    Surface studies of the northern lowlands of Mars have shown that this region has undergone a complex history including volcanism, sedimentary deposition and secondary modification by climate change. Despite these analyses, the origin and the evolution of this region are still debated. No clear and definitive evidences have been found so far to conclude whether these plains were formed by a giant impact, were once covered by an ocean or were filled by a large quantity of lavas. In the visible and infrared spectral range, the northern lowlands differ from southern terrains in the NIR negative slope while they exhibit VNIR spectra similar to the southern pyroxene-rich areas (Carrozzo et al., 2012). These observations, combined with both recent detection of mafic minerals at higher spatial resolution by CRISM (Salvatore et al., 2010) and recent results of Horgan and Bell (2012), supports that their mineralogy is linked to weathered basalts with a glassy component. In addition to this, the spectral similarity of Acidalia area with the northern circumpolar sand dunes, apart from the hydration features, suggests that the weathering processes that took place there could be related to past glacial activity, in agreement with superficial morphology showing glacial structures. Aim of this work is to combine the OMEGA mineralogical maps with morphological features (Tanaka et al., 2011) and investigate possible terrestrial analogues in order to give some constrains on the composition and origin of these puzzling Martian terrains.

  10. Tourism, biodiversity and protected areas--Review from northern Fennoscandia.

    PubMed

    Tolvanen, Anne; Kangas, Katja

    2016-03-15

    Tourist numbers in northern Fennoscandia outweigh those in other northern boreal - arctic regions, which creates a specific need to evaluate the impacts of tourism. This review 1) identifies patterns and trends in the vegetation and wildlife of northern Fennoscandian terrestrial ecosystems as a consequence of tourism and recreation, 2) discusses the implications of findings in terms of the intensity, area and magnitude of impacts, changing climate and management needs under increasing tourist pressure, and 3) identifies research gaps. The reviewed studies show negative environmental and biodiversity impacts that are most pronounced near tourist resorts. The most sensitive plants, birds and mammals decline or disappear from the disturbed sites, and the species composition shifts from 'wild' species to cultural and human associated species. There is little research on the spread of alien species, but the few examples show that alien species can be promoted by tourism activities. Impacts of the use of motorized vehicles have not been widely studied either, despite the extensive track network which can cause disturbance to wildlife. The integrated impacts of tourism and climate change on the vegetation and wildlife was not addressed directly in any of the reviewed studies. In addition, little research has been done on carrying out restoration at tourist areas. Scientific research on these topics is needed to prevent, minimize or restore the most negative ecological impacts of tourism and recreation. PMID:26720330

  11. Comparison of Two Widely Used Human Papillomavirus Detection and Genotyping Methods, GP5+/6+-Based PCR Followed by Reverse Line Blot Hybridization and Multiplex Type-Specific E7-Based PCR.

    PubMed

    Clifford, Gary M; Vaccarella, Salvatore; Franceschi, Silvia; Tenet, Vanessa; Umulisa, M Chantal; Tshomo, Ugyen; Dondog, Bolormaa; Vorsters, Alex; Tommasino, Massimo; Heideman, Daniëlle A M; Snijders, Peter J F; Gheit, Tarik

    2016-08-01

    GP5+/6+-based PCR followed by reverse line blot hybridization (GP5+/6+RLB) and multiplex type-specific PCR (E7-MPG) are two human papillomavirus (HPV) genotyping methodologies widely applied in epidemiological research. We investigated their relative analytical performance in 4,662 samples derived from five studies in Bhutan, Rwanda, and Mongolia coordinated by the International Agency for Research on Cancer (IARC). A total of 630 samples were positive by E7-MPG only (13.5%), 24 were positive by GP5+/6+RLB only (0.5%), and 1,014 were positive (21.8%) by both methods. Ratios of HPV type-specific positivity of the two tests (E7-MPG:GP5+/6+RLB ratio) were calculated among 1,668 samples that were HPV positive by one or both tests. E7-MPG:GP5+/6+RLB ratios were >1 for all types and highly reproducible across populations and sample types. E7-MPG:GP5+/6+RLB ratios were highest for HPV53 (7.5) and HPV68 (7.1). HPV16 (1.6) and HPV18 (1.7) had lower than average E7-MPG:GP5+/6+RLB ratios. Among E7-MPG positive infections, median mean fluorescence intensity (MFI; a semiquantitative measure of viral load) tended to be higher among samples positive for the same virus type by GP5+/6+RLB than for those negative for the same type by GP5+/6+RLB. Exceptions, however, included HPV53, -59, and -82, for which the chances of being undetected by GP5+/6+RLB appeared to be MFI independent. Furthermore, the probability of detecting an additional type by E7-MPG was higher when another type was already detected by GP5+/6+RLB, suggesting the existence of masking effects due to competition for GP5+/6+ PCR primers. In conclusion, this analysis is not an evaluation of clinical performance but may inform choices for HPV genotyping methods in epidemiological studies, when the relative merits and dangers of sensitivity versus specificity for individual types should be considered, as well as the potential to unmask nonvaccine types following HPV vaccination. PMID:27225411

  12. Northern Terra Meridiani's 'Monument Valley'

    NASA Technical Reports Server (NTRS)

    2002-01-01

    Northern Terra Meridiani, near the intersection of the martian equator and prime meridian, is a region of vast exposures of layered rock. A thermal image from the Phobos 2 orbiter in 1989 showed these materials to be anomalously cool during the daytime, an observation very suggestive of dense, hardened materials like rock. Mars Global Surveyor (MGS) Mars Orbiter Camera (MOC) images of this region show layered material exposed in cliffs, buttes, and mesas that in some ways resemble the rock outcrops of northern Arizona and southeastern Utah in North America (e.g., Monument Valley, Canyonlands, Zion National Park, Four Corners). MGS MOC Extended Mission operations have included several hundred opportunities for the spacecraft to be rolled off-nadir (i.e., at an angle other than 'straight down') to take pictures that repeat earlier MOC coverage. These repeat images, because they are taken from a different angle, can be combined with the original picture to produce a stereoscopic ('3-D') view. The image shown here is a composite of two pictures, the first taken October 23, 2000, the second acquired by pointing the spacecraft off-nadir on May 15, 2001. This view shows four buttes and a pinnacle (near left-center) composed of eroded, layered rock. The four buttes are each capped by the remains of a single layer of rock that is harder than the materials beneath it. It is the presence of this cap rock that has permitted these buttes to remain standing after surrounding materials were eroded away. Like the buttes of Monument Valley in the Navajo Nation on the Arizona/Utah border, these are believed to consist of sedimentary rocks, perhaps deposited in water or by wind, though some scientists have speculated that they could be made of thick accumulations of volcanic ash. The image covers an area approximately 3 km (1.9 mi) across and is illuminated by sunlight from the left. To see the image in 3-D, red (left-eye) and blue (right-eye) '3-D' glasses are required.

  13. Bioactive compounds from northern plants.

    PubMed

    Hohtola, Anja

    2010-01-01

    Northern conditions are characterised by long days with much light and low temperatures during the growing season. It has been chimed that herbs and berries grown in the north are stronger tasting compared to those of southern origin. The compounds imparting aroma and color to berries and herbs are secondary metabolites which in plants mostly act as chemical means of defense. Recently, the production of secondary metabolites using plant cells has been the subject of expanding research. Light intensity, photoperiod and temperature have been reported to influence the biosynthesis of many secondary metabolites. Native wild aromatic and medicinal plant species of different families are being studied to meet the needs of raw material for the expanding industry of e.g., health-promoting food products known as nutraceutics. There are already a large number of known secondary compounds produced by plants, but the recent advances in modern extraction and analysis should enable many more as yet unknown compounds to be found, characterised and utilised. Rose root (Rhodiola rosea) is a perennial herbaceous plant which inhabits mountain regions throughout Europe, Asia and east coastal regions of North America. The extract made from the rhizomes acts as a stimulant like the Ginseng root. Roseroot has been categorized as an adaptogen and is reported to have many pharmacological properties. The biologically active components of the extract are salitroside tyrosol and cinnamic acid glycosides (rosavin, rosarin, rosin). Round-leaved sundew (Drosera rotundifolia L.) has circumboreal distribution. It inhabits nutrient-poor, moist and sunny areas such as peat bogs and wetlands. Sundew leaves are collected from the wild-type for various medicinal preparations and can be utilized in treating e.g., as an important "cough-medicine" for different respiratory diseases. The antimicrobial activity of extracts of aerial parts against various bacteria has been investigated. Drosera produces

  14. Energetics and growth rate of northern Shrike (Lanius excubitor) nestlings

    SciTech Connect

    Degen, A.A.; Kam, M. ); Pinshow, B.; Yosef, R. ); Nagy, K.A. )

    1992-12-01

    Northern Shrikes (Lanius excubitor) breed in a variety of habitats, including deserts. Deserts are characterized by unpredictable food supplies, which can lead to a slow growth rate of nestlings. However, given that Northern Shrike males use prey from their caches to augment freshly caught prey in providing food for their mates and nestlings, we hypothesized that their nestlings do not have a slow growth rate, but one that is equivalent to that in other passerine nestlings from temperate areas. To test this hypothesis, we measured growth rates and energy use in Northern Shrike nestlings and fledglings. We also measured energy expenditure in two adult males that were attending nests. Growth rate of Northern Shrike nestlings was similar to that predicted for passerines in temperate areas and therefore our hypothesis was supported. However, metabolizable energy available in the cache amounted to only [approx] 7.2% of the total energy requirements of the nestlings or 4.2% of the total energy requirements of parents and nestlings during the nestling period. This suggested that other factors in addition to the cache were important in determining growth rate. These included (1) an extremely low maintenance energy requirement of the nestling; 30% of that predicted for a bird of its body mass when it weighed 10 g, which gradually increased to 70% at 50 g. This allowed for more of the energy intake to be used for growth and also reduced foraging costs of males; (2) the relatively low amount of body energy retained as a fraction of metabolizable energy intake, 0.15 to 0.16, indicating that more water per unit growth was incorporated than in other passerines. 47 refs., 5 figs., 2 tabs.

  15. The Northern Bering Sea: An Arctic Ecosystem in Change

    NASA Astrophysics Data System (ADS)

    Grebmeier, J. M.; Cooper, L. W.

    2004-12-01

    Arctic systems can be rich and diverse habitats for marine life in spite of the extreme cold environment. Benthic faunal populations and associated biogeochemical cycling processes are influenced by sea-ice extent, seawater hydrography (nutrients, salinity, temperature, currents), and water column production. Benthic organisms on the Arctic shelves and margins are long-term integrators of overlying water column processes. Because these organisms have adapted to living at cold extremes, it is reasonable to expect that these communities will be among the most susceptible to climate warming. Recent observations show that Arctic sea ice in the North American Arctic is melting and retreating northward earlier in the season and the timing of these events can have dramatic impacts on the biological system. Changes in overlying primary production, pelagic-benthic coupling, and benthic production and community structure can have cascading effects to higher trophic levels, particularly benthic feeders such as walruses, gray whales, and diving seaducks. Recent indicators of contemporary Arctic change in the northern Bering Sea include seawater warming and reduction in ice extent that coincide with our time-series studies of benthic clam population declines in the shallow northern Bering shelf in the 1990's. In addition, declines in benthic amphipod populations have also likely influenced the movement of feeding gray whales to areas north of Bering Strait during this same time period. Finally a potential consequence of seawater warming and reduced ice extent in the northern Bering Sea could be the northward movement of bottom feeding fish currently in the southern Bering Sea that prey on benthic fauna. This would increase the feeding pressure on the benthic prey base and enhance competition for this food source for benthic-feeding marine mammals and seabirds. This presentation will outline recent biological changes observed in the northern Bering Sea ecosystem as documented in

  16. Genetically distinct coelacanth population off the northern Tanzanian coast

    PubMed Central

    Nikaido, Masato; Sasaki, Takeshi; Emerson, J. J.; Aibara, Mitsuto; Mzighani, Semvua I.; Budeba, Yohana L.; Ngatunga, Benjamin P.; Iwata, Masamitsu; Abe, Yoshitaka; Li, Wen-Hsiung; Okada, Norihiro

    2011-01-01

    Since the sensational discovery of a living coelacanth off the east coast of South Africa, the geographic distribution of viable coelacanth populations has been a subject of debate. In the past, the coelacanths off the African mainland were thought to be strays from the Comoros because most coelacanths captured were caught in the waters surrounding the Comoros archipelagos. However, in recent years, a large number of coelacanths were captured off the coast of Tanzania, including nine living specimens observed in a remotely operated vehicles survey. Thus, it is possible that there is a reproducing population inhabiting waters off the Tanzania coast. We have sequenced the complete mitochondrial genomes of 21 Tanzanian and 2 Comoran coelacanths and analyzed these sequences together with two additional full mitochondrial genomes and 47 d-loop sequences from the literature. We found that the coelacanth population off the northern Tanzanian coast is genetically differentiated from those of the southern Tanzania coast and the Comoros, whereas no significant genetic differentiation occurs between the latter two localities. The differentiation between the northern and southern Tanzanian coast populations is consistent with the hypothesis that the existence of northward-flowing ocean current along the Tanzanian coast may reduce or prevent gene flow from the northern to the southern population. Finally, we estimated that the population localized to the southern Tanzanian coast and the Comoros diverged from other coelacanths at least 200,000 y ago. These results indicate that the coelacanths off the northern Tanzania coast are not strays but a genetically distinct group. Our study provides important information for the conservation of this threatened “living fossil.” PMID:22025696

  17. Decomposition in northern Minnesota peatlands

    SciTech Connect

    Farrish, K.W.

    1985-01-01

    Decomposition in peatlands was investigated in northern Minnesota. Four sites, an ombrotrophic raised bog, an ombrotrophic perched bog and two groundwater minerotrophic fens, were studied. Decomposition rates of peat and paper were estimated using mass-loss techniques. Environmental and substrate factors that were most likely to be responsible for limiting decomposition were monitored. Laboratory incubation experiments complemented the field work. Mass-loss over one year in one of the bogs, ranged from 11 percent in the upper 10 cm of hummocks to 1 percent at 60 to 100 cm depth in hollows. Regression analysis of the data for that bog predicted no mass-loss below 87 cm. Decomposition estimates on an area basis were 2720 and 6460 km/ha yr for the two bogs; 17,000 and 5900 kg/ha yr for the two fens. Environmental factors found to limit decomposition in these peatlands were reducing/anaerobic conditions below the water table and cool peat temperatures. Substrate factors found to limit decomposition were low pH, high content of resistant organics such as lignin, and shortages of available N and K. Greater groundwater influence was found to favor decomposition through raising the pH and perhaps by introducing limited amounts of dissolved oxygen.

  18. Northern California near San Francisco

    NASA Technical Reports Server (NTRS)

    1974-01-01

    A part of northern California centered near San Francisco Bay (38.0N, 122.0W) photographed at 3 p.m. January 1, 1974, from the Skylab space station in Earth orbit. This near vertical view encompasses the coastline from Monteray Bay (right) to about 50 miles north of Point Reyes (left) and includes, from bottom to top, San Francisco Bay (center), Sacramento Valley (left center), San Joaquin Valley (right center), and the snow-covered Sierra Nevada. Afternoon shadows sharply delineate a valley which parallels San Francisco Bay, crosses Point Reyes, and lies between the Bay and the Pacific coastline. This valley marks the location of the San Andreas Fault. Forces acting on the crust are causing the land west (bottom) of the fault line to move north relative to land on the east side. Agricultural areas in the Sacramento and San Joaquin Valleys are indicated by the tan areas which are easily discerned in contrast to the green-gray background.

  19. Dilution of the northern North Atlantic Ocean in recent decades.

    PubMed

    Curry, Ruth; Mauritzen, Cecilie

    2005-06-17

    Declining salinities signify that large amounts of fresh water have been added to the northern North Atlantic Ocean since the mid-1960s. We estimate that the Nordic Seas and Subpolar Basins were diluted by an extra 19,000 +/- 5000 cubic kilometers of freshwater input between 1965 and 1995. Fully half of that additional fresh water-about 10,000 cubic kilometers-infiltrated the system in the late 1960s at an approximate rate of 2000 cubic kilometers per year. Patterns of freshwater accumulation observed in the Nordic Seas suggest a century time scale to reach freshening thresholds critical to that portion of the Atlantic meridional overturning circulation. PMID:15961666

  20. Enantioselective Michael Addition of Water

    PubMed Central

    Chen, Bi-Shuang; Resch, Verena; Otten, Linda G; Hanefeld, Ulf

    2015-01-01

    The enantioselective Michael addition using water as both nucleophile and solvent has to date proved beyond the ability of synthetic chemists. Herein, the direct, enantioselective Michael addition of water in water to prepare important β-hydroxy carbonyl compounds using whole cells of Rhodococcus strains is described. Good yields and excellent enantioselectivities were achieved with this method. Deuterium labeling studies demonstrate that a Michael hydratase catalyzes the water addition exclusively with anti-stereochemistry. PMID:25529526

  1. Gasoline additives, emissions, and performance

    SciTech Connect

    1995-12-31

    The papers included in this publication deal with the influence of fuel, additive, and hardware changes on a variety of vehicle performance characteristics. Advanced techniques for measuring these performance parameters are also described. Contents include: Fleet test evaluation of gasoline additives for intake valve and combustion chamber deposit clean up; A technique for evaluating octane requirement additives in modern engines on dynamometer test stands; A fleet test of two additive technologies comparing their effects on tailpipe emissions; Investigation into the vehicle exhaust emissions of high percentage ethanol blends; Variability in hydrocarbon speciation measurements at low emission (ULEV) levels; and more.

  2. Long-term variability in Northern Hemisphere snow cover and associations with warmer winters

    USGS Publications Warehouse

    McCabe, G.J.; Wolock, D.M.

    2010-01-01

    A monthly snow accumulation and melt model is used with gridded monthly temperature and precipitation data for the Northern Hemisphere to generate time series of March snow-covered area (SCA) for the period 1905 through 2002. The time series of estimated SCA for March is verified by comparison with previously published time series of SCA for the Northern Hemisphere. The time series of estimated Northern Hemisphere March SCA shows a substantial decrease since about 1970, and this decrease corresponds to an increase in mean winter Northern Hemisphere temperature. The increase in winter temperature has caused a decrease in the fraction of precipitation that occurs as snow and an increase in snowmelt for some parts of the Northern Hemisphere, particularly the mid-latitudes, thus reducing snow packs and March SCA. In addition, the increase in winter temperature and the decreases in SCA appear to be associated with a contraction of the circumpolar vortex and a poleward movement of storm tracks, resulting in decreased precipitation (and snow) in the low- to mid-latitudes and an increase in precipitation (and snow) in high latitudes. If Northern Hemisphere winter temperatures continue to warm as they have since the 1970s, then March SCA will likely continue to decrease. ?? 2009 Springer Science+Business Media B.V.

  3. Long-term variability in Northern Hemisphere snow cover and associations with warmer winters

    USGS Publications Warehouse

    McCabe, Gregory J.; Wolock, David M.

    2010-01-01

    A monthly snow accumulation and melt model is used with gridded monthly temperature and precipitation data for the Northern Hemisphere to generate time series of March snow-covered area (SCA) for the period 1905 through 2002. The time series of estimated SCA for March is verified by comparison with previously published time series of SCA for the Northern Hemisphere. The time series of estimated Northern Hemisphere March SCA shows a substantial decrease since about 1970, and this decrease corresponds to an increase in mean winter Northern Hemisphere temperature. The increase in winter temperature has caused a decrease in the fraction of precipitation that occurs as snow and an increase in snowmelt for some parts of the Northern Hemisphere, particularly the mid-latitudes, thus reducing snow packs and March SCA. In addition, the increase in winter temperature and the decreases in SCA appear to be associated with a contraction of the circumpolar vortex and a poleward movement of storm tracks, resulting in decreased precipitation (and snow) in the low- to mid-latitudes and an increase in precipitation (and snow) in high latitudes. If Northern Hemisphere winter temperatures continue to warm as they have since the 1970s, then March SCA will likely continue to decrease.

  4. LEECHES (ANNELIDA: EUHIRUDINEA) OF NORTHERN ARKANSAS

    EPA Science Inventory

    Twenty-one lotic and lentic environments throughout central and northern Arkansas were surveyed for the presence of leeches during June 2004, and April, July - October, 2005. Fourteen species of leeches (Desserobdella cryptobranchii, Desserobdella phalera, Desserobdella picta, H...

  5. Huge Filament Rises From Sun's Northern Hemisphere

    NASA Video Gallery

    On August 1, 2010 following a C3-class solar flare from sunspot 1092, an enormous magnetic filament stretching across the sun's northern hemisphere erupted. This 304 angstrom video shows that filam...

  6. Growth Pattern Responses to Photoperiod across Latitudes in a Northern Damselfly

    PubMed Central

    Śniegula, Szymon; Nilsson-Örtman, Viktor; Johansson, Frank

    2012-01-01

    Background Latitudinal clines in temperature and seasonality impose strong seasonal constraints on ectotherms. Studies of population differentiation in phenotypic plasticity of life history traits along latitudinal gradients are important for understanding how organisms have adapted to seasonal environments and predict how they respond to climate changes. Such studies have been scarce for species with a northern distribution. Methodology/Principle Finding Larvae of the northern damselfly Coenagrion johanssoni originating from semivoltine central, partivoltine northern, and partivoltine northernmost Swedish populations were reared in the laboratory. To investigate whether larvae use photoperiodic cues to induce compensatory growth along this latitudinal gradient, larvae were reared under two different photoperiods corresponding to a northern and southern latitude. In addition, field adult size was assessed to test the strength of possible compensatory growth mechanisms under natural conditions and hatchling size was measured to test for maternal effects. We hypothesized that populations originating from lower latitudes would be more time constrained than high-latitude populations because they have a shorter life cycle. The results showed that low-latitude populations had higher growth rates in summer/fall. In general northern photoperiods induced higher growth rates, but this plastic response to photoperiod was strongest in the southernmost populations and negligible in the northernmost population. During spring, central populations grew faster under the southern rather than the northern photoperiod. On the other hand, northern and northernmost populations did not differ between each other and grew faster in the northern rather than in the southern photoperiod. Field sampled adults did not differ in size across the studied regions. Conclusion/Significance We found a significant differentiation in growth rate across latitudes and latitudinal difference in growth rate

  7. Patterns and Risks of Trichinella Infection in Humans and Pigs in Northern Laos

    PubMed Central

    Conlan, James V.; Vongxay, Khamphouth; Khamlome, Boualam; Gomez-Morales, Maria Angeles; Pozio, Edoardo; Blacksell, Stuart D.; Fenwick, Stanley; Thompson, R. C. A.

    2014-01-01

    Several outbreaks of trichinellosis associated with the consumption of raw pork have occurred in Laos since 2004. This cross-sectional study was conducted in four provinces of northern Laos to investigate the seroepidemiology of trichinellosis in the human population and determine the prevalence and species of Trichinella infection in the domestic pig population. Serum samples and questionnaire data were obtained from 1419 individuals. Serum samples were tested for Trichinella antibodies by ELISA using larval excretory–secretory (ES) antigens and a subset of 68 positive samples were tested by western blot. The seroprevalence of Trichinella antibodies was 19.1% (95% confidence interval (CI) = 17.1–21.1%). The risk of having antibodies detected by ELISA using ES antigens increased with age, being of Lao-Tai ethnicity, living in Oudomxay province and being male. Tongue and diaphragm muscle samples were collected from 728 pigs and tested for Trichinella larvae by the artificial digestion method. Trichinella larvae were isolated from 15 pigs (2.1%) of which 13 were identified as T. spiralis by molecular typing; the species of the two remaining isolates could not be determined due to DNA degradation. Trichinella spp. are endemic in the domestic environment of northern Laos and targeted preventative health measures should be initiated to reduce the risk of further outbreaks occurring. PMID:25078088

  8. Color Addition and Subtraction Apps

    ERIC Educational Resources Information Center

    Ruiz, Frances; Ruiz, Michael J.

    2015-01-01

    Color addition and subtraction apps in HTML5 have been developed for students as an online hands-on experience so that they can more easily master principles introduced through traditional classroom demonstrations. The evolution of the additive RGB color model is traced through the early IBM color adapters so that students can proceed step by step…

  9. 75 FR 27313 - Proposed Additions

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-05-14

    ... FROM PEOPLE WHO ARE BLIND OR SEVERELY DISABLED PROCUREMENT LIST Proposed Additions AGENCY: Committee for Purchase From People Who Are Blind or Severely Disabled. ACTION: Proposed additions to the... or Severely Disabled, Jefferson Plaza 2, Suite 10800, 1421 Jefferson Davis Highway,...

  10. Northern Taurids in the IAU MDC Database

    NASA Astrophysics Data System (ADS)

    Kaňuchová, Z.; Svoreň, J.

    2012-11-01

    The method of indices was used to study the northern branch of the autumn (night) part of the Taurid complex. The procedure based only on mathematical statistics was applied to select the Northern Taurid meteor records from the IAU Meteor Data Center Database. Because we wanted to study especially the structure of the inner part of the Northern Taurids, we were focused on the interval of the higher activity of the stream — from the end of the Perseids activity to the beginning of the Geminids activity. We did not take into account outlying parts of the complex, which is active, according to some authors, until January. 84 orbits of the Northern Taurids were selected. 63 of 84 Northern Taurids orbits (75%) were sorted into 11 associations found in the stream. One of the associations consisting of three orbits was identified as a previously unknown northern branch of τ Arietids shower. We also found an association with orbital characteristics equal to the characteristics of showers δ Psc N and χ Ori N. Meteors in these associations were observed up to three weeks earlier compared to currently cataloged data of the showers. The orientation of the mean orbit of a 5-member association of δ Psc N, different from the general trend, indicates that this stream may not be genetically related to other members of the Taurid complex.

  11. 40 CFR 81.354 - Northern Mariana Islands.

    Code of Federal Regulations, 2014 CFR

    2014-07-01

    ... 40 Protection of Environment 18 2014-07-01 2014-07-01 false Northern Mariana Islands. 81.354... § 81.354 Northern Mariana Islands. Northern Mariana Islands—TSP Designated area Does not meet primary... 1 X 1 EPA designation only. Northern Mariana Islands—1971 Sulfur Dioxide NAAQS (Primary...

  12. 75 FR 41842 - Northern Border Pipeline Company; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-07-19

    ... Energy Regulatory Commission Northern Border Pipeline Company; Notice of Application July 12, 2010. Take notice that on July 2, 2010, Northern Border Pipeline Company (Northern Border), 717 Texas Street... Northern Border's mainline system in Bureau County, Illinois, to a point of interconnection with...

  13. Identification of immunodominant VP1 linear epitope of enterovirus 71 (EV71) using synthetic peptides for detecting human anti-EV71 IgG antibodies in Western blots.

    PubMed

    Foo, D G W; Ang, R X; Alonso, S; Chow, V T K; Quak, S H; Poh, C L

    2008-03-01

    A major IgG-specific immunodominant VP1 linear epitope of enterovirus 71 (EV71) strain 41 (5865/SIN/00009), defined by the core sequence LEGTTNPNG, was identified by Pepscan analysis. Oligonucleotides corresponding to the amino-acid sequence of synthetic peptide SP32 were cloned and over-expressed in Escherichia coli as a recombinant glutathione-S-transferase (GST)-SP32 fusion protein. In ELISAs, this protein did not react with human anti-EV71 IgG antibodies, but there was significant immunoreactivity according to western blot analysis. The amino-acid sequence of SP32 was highly specific for detecting EV71 strains in western blot analysis, and showed no immunoreactivity with monoclonal antibodies raised against other enteroviruses, e.g., CA9 and Echo 6. PMID:18076666

  14. Seismic risk assessment of Navarre (Northern Spain)

    NASA Astrophysics Data System (ADS)

    Gaspar-Escribano, J. M.; Rivas-Medina, A.; García Rodríguez, M. J.; Benito, B.; Tsige, M.; Martínez-Díaz, J. J.; Murphy, P.

    2009-04-01

    The RISNA project, financed by the Emergency Agency of Navarre (Northern Spain), aims at assessing the seismic risk of the entire region. The final goal of the project is the definition of emergency plans for future earthquakes. With this purpose, four main topics are covered: seismic hazard characterization, geotechnical classification, vulnerability assessment and damage estimation to structures and exposed population. A geographic information system is used to integrate, analyze and represent all information colleted in the different phases of the study. Expected ground motions on rock conditions with a 90% probability of non-exceedance in an exposure time of 50 years are determined following a Probabilistic Seismic Hazard Assessment (PSHA) methodology that includes a logic tree with different ground motion and source zoning models. As the region under study is located in the boundary between Spain and France, an effort is required to collect and homogenise seismological data from different national and regional agencies. A new homogenised seismic catalogue, merging data from Spanish, French, Catalonian and international agencies and establishing correlations between different magnitude scales, is developed. In addition, a new seismic zoning model focused on the study area is proposed. Results show that the highest ground motions on rock conditions are expected in the northeastern part of the region, decreasing southwards. Seismic hazard can be expressed as low-to-moderate. A geotechnical classification of the entire region is developed based on surface geology, available borehole data and morphotectonic constraints. Frequency-dependent amplification factors, consistent with code values, are proposed. The northern and southern parts of the region are characterized by stiff and soft soils respectively, being the softest soils located along river valleys. Seismic hazard maps including soil effects are obtained by applying these factors to the seismic hazard maps

  15. Multiple Landslide-Hazard Scenarios Modeled for the Oakland-Berkeley Area, Northern California

    USGS Publications Warehouse

    Pike, Richard J.; Graymer, Russell W.

    2008-01-01

    With the exception of Los Angeles, perhaps no urban area in the United States is more at risk from landsliding, triggered by either precipitation or earthquake, than the San Francisco Bay region of northern California. By January each year, seasonal winter storms usually bring moisture levels of San Francisco Bay region hillsides to the point of saturation, after which additional heavy rainfall may induce landslides of various types and levels of severity. In addition, movement at any time along one of several active faults in the area may generate an earthquake large enough to trigger landslides. The danger to life and property rises each year as local populations continue to expand and more hillsides are graded for development of residential housing and its supporting infrastructure. The chapters in the text consist of: *Introduction by Russell W. Graymer *Chapter 1 Rainfall Thresholds for Landslide Activity, San Francisco Bay Region, Northern California by Raymond C. Wilson *Chapter 2 Susceptibility to Deep-Seated Landsliding Modeled for the Oakland-Berkeley Area, Northern California by Richard J. Pike and Steven Sobieszczyk *Chapter 3 Susceptibility to Shallow Landsliding Modeled for the Oakland-Berkeley Area, Northern California by Kevin M. Schmidt and Steven Sobieszczyk *Chapter 4 Landslide Hazard Modeled for the Cities of Oakland, Piedmont, and Berkeley, Northern California, from a M=7.1 Scenario Earthquake on the Hayward Fault Zone by Scott B. Miles and David K. Keefer *Chapter 5 Synthesis of Landslide-Hazard Scenarios Modeled for the Oakland-Berkeley Area, Northern California by Richard J. Pike The plates consist of: *Plate 1 Susceptibility to Deep-Seated Landsliding Modeled for the Oakland-Berkeley Area, Northern California by Richard J. Pike, Russell W. Graymer, Sebastian Roberts, Naomi B. Kalman, and Steven Sobieszczyk *Plate 2 Susceptibility to Shallow Landsliding Modeled for the Oakland-Berkeley Area, Northern California by Kevin M. Schmidt and Steven

  16. Color Addition and Subtraction Apps

    NASA Astrophysics Data System (ADS)

    Ruiz, Frances; Ruiz, Michael J.

    2015-10-01

    Color addition and subtraction apps in HTML5 have been developed for students as an online hands-on experience so that they can more easily master principles introduced through traditional classroom demonstrations. The evolution of the additive RGB color model is traced through the early IBM color adapters so that students can proceed step by step in understanding mathematical representations of RGB color. Finally, color addition and subtraction are presented for the X11 colors from web design to illustrate yet another real-life application of color mixing.

  17. Selliguea kachinensis (Polypodiaceae), a new fern species of uncertain affinity from Northern Myanmar.

    PubMed

    Khine, Phyo Kay; Lindsay, Stuart; Fraser-Jenkins, Christopher; Kluge, Jürgen; Kyaw, Myint; Hovenkamp, Peter

    2016-01-01

    We describe Selliguea kachinensis as a new species from Northern Myanmar and discuss its generic placement in either Selliguea or Arthromeris. The conservation status is assessed as Data Deficient. In addition, we make the new combination Selliguea erythrocarpa (Mett. ex Kuhn) Hovenkamp, S. Linds., Fraser-Jenk. PMID:27212883

  18. Advanced Technology Tech Prep Partnership for Northern Kane Regional Delivery System. Final Report.

    ERIC Educational Resources Information Center

    Elgin Community Coll., IL.

    A 1-year project was undertaken to continue implementation, evaluation, and revision of a model advanced technology partnership between Elgin Community College (ECC) and the Northern Kane Regional Delivery System in Illinois. The model program, which originally included three high schools, was expanded to include five additional high schools in…

  19. Selliguea kachinensis (Polypodiaceae), a new fern species of uncertain affinity from Northern Myanmar

    PubMed Central

    Khine, Phyo Kay; Lindsay, Stuart; Fraser-Jenkins, Christopher; Kluge, Jürgen; Kyaw, Myint; Hovenkamp, Peter

    2016-01-01

    Abstract We describe Selliguea kachinensis as a new species from Northern Myanmar and discuss its generic placement in either Selliguea or Arthromeris. The conservation status is assessed as Data Deficient. In addition, we make the new combination Selliguea erythrocarpa (Mett. ex Kuhn) Hovenkamp, S. Linds., Fraser-Jenk. PMID:27212883

  20. Calculators and Computers: Graphical Addition.

    ERIC Educational Resources Information Center

    Spero, Samuel W.

    1978-01-01

    A computer program is presented that generates problem sets involving sketching graphs of trigonometric functions using graphical addition. The students use calculators to sketch the graphs and a computer solution is used to check it. (MP)

  1. Polyolefins as additives in plastics

    SciTech Connect

    Deanin, R.D.

    1993-12-31

    Polyolefins are not only major commodity plastics - they are also very useful as additives, both in other polyolefins and also in other types of plastics. This review covers ethylene, propylene, butylene and isobutylene polymers, in blends with each other, and as additives to natural rubber, styrene/butadiene rubber, polystyrene, polyvinyl chloride, polymethyl methacrylate, polyphenylene oxide, polycarbonate, thermoplastic polyesters, polyurethanes, polyamides, and mixed automotive plastics recycling.

  2. Psoas abscess diagnosed at a Northern university hospital.

    PubMed

    Maagaard, Anne; Oktedalen, Olav

    2002-01-01

    Abscess of the psoas muscle is an infrequent diagnosis at hospitals in Northern countries. We report on 16 patients who had this diagnosis during the period 1991-2001. Eight patients were immigrants who had previously been healthy and most of them had experienced symptoms for approximately 1 y. MRI or CT scans revealed spondylodiscitis in 6 of these patients and Mycobacterium tuberculosis was identified as the causative agent. With the exception of 1 patient who was exclusively treated with antituberculous agents, all 8 immigrant patients were successfully treated with antituberculous agents in addition to percutaneous drainage. The other 8 patients were Norwegians, 4 of whom had underlying conditions such as diabetes mellitus or drug abuse. The causative microorganisms were Staphylococcus aureus or Streptococcus spp., with the exception of M. tuberculosis in 1 case. The Norwegian patients had a more acute history of symptoms than the immigrant patients and 2 of them were in a septic condition on admittance. Two of the Norwegians died of serious infection; 5 were successfully treated with percutaneous drainage in addition to antibiotics and 1 was treated exclusively with antibiotic agents. The clinical history and microorganism associated with psoas abscess seemed to depend on whether or not the patient was an immigrant. Owing to increasing immigration, diagnosis of psoas abscess should be taken into account in Northern countries. PMID:12578159

  3. Variations in the Teleconnection of ENSO and Summer Rainfall in Northern China: A Role of the Indian Summer Monsoon(.

    NASA Astrophysics Data System (ADS)

    Feng, Song; Hu, Qi

    2004-12-01

    Observational studies have created a dilemma on how El Niño Southern Oscillation (ENSO) may have affected interannual variations of summer rainfall in northern China; some suggested a consistent effect while others showed a complete lack of effect. This dilemma is resolved in this study, which shows that ENSO has affected the summer rainfall in northern China and the effect has varied at multidecadal scales. The question of how the ENSO teleconnection with northern China rainfall variation was established is addressed, and an answer pointing to the Indian summer monsoon as a “facilitator” connecting ENSO and northern China rainfall variation is examined. The Indian monsoon circulation interacted with the regional circulations in northern China in some epochs and such interaction was interrupted in other epochs. When the interaction was active, the Indian monsoon variations originating from ENSO, during El Niño or La Niña, was extended to affect the rainfall variation in northern China, creating a teleconnection of ENSO with northern China rainfall. When the interaction weakened or was inactive, the ENSO effect languished. Additional analyses were done to address the related question of why the interactions have alternated. The alternation was suggested to result from variations of the large-scale circulation in the Eurasian continent. The circulation anomalies showed lowering (rising) 500-hPa geopotential height centered at Mongolia and western China in some epochs, enhancing cyclonic (anticyclonic) rotation in mid- and low-level winds and creating (disrupting) a moisture convoy from the Indian monsoon region to northern China and synergetic convergence/divergence anomalies in the monsoon region and in northern China. Results of this study contribute to the understanding of interannual and multidecadal variations of the summer rainfall in the semiarid region of northern China.


  4. ADDITIVITY ASSESSMENT OF TRIHALOMETHANE MIXTURES BY PROPORTIONAL RESPONSE ADDITION

    EPA Science Inventory

    If additivity is known or assumed, the toxicity of a chemical mixture may be predicted from the dose response curves of the individual chemicals comprising the mixture. As single chemical data are abundant and mixture data sparse, mixture risk methods that utilize single chemical...

  5. [INVITED] Lasers in additive manufacturing

    NASA Astrophysics Data System (ADS)

    Pinkerton, Andrew J.

    2016-04-01

    Additive manufacturing is a topic of considerable ongoing interest, with forecasts predicting it to have major impact on industry in the future. This paper focusses on the current status and potential future development of the technology, with particular reference to the role of lasers within it. It begins by making clear the types and roles of lasers in the different categories of additive manufacturing. This is followed by concise reviews of the economic benefits and disadvantages of the technology, current state of the market and use of additive manufacturing in different industries. Details of these fields are referenced rather than expanded in detail. The paper continues, focusing on current indicators to the future of additive manufacturing. Barriers to its development, trends and opportunities in major industrial sectors, and wider opportunities for its development are covered. Evidence indicates that additive manufacturing may not become the dominant manufacturing technology in all industries, but represents an excellent opportunity for lasers to increase their influence in manufacturing as a whole.

  6. Evaluation of certain food additives.

    PubMed

    2015-01-01

    This report represents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of various food additives, including flavouring agents, and to prepare specifications for identity and purity. The first part of the report contains a general discussion of the principles governing the toxicological evaluation of and assessment of dietary exposure to food additives, including flavouring agents. A summary follows of the Committee's evaluations of technical, toxicological and dietary exposure data for eight food additives (Benzoe tonkinensis; carrageenan; citric and fatty acid esters of glycerol; gardenia yellow; lutein esters from Tagetes erecta; octenyl succinic acid-modified gum arabic; octenyl succinic acid-modified starch; paprika extract; and pectin) and eight groups of flavouring agents (aliphatic and alicyclic hydrocarbons; aliphatic and aromatic ethers; ionones and structurally related substances; miscellaneous nitrogen-containing substances; monocyclic and bicyclic secondary alcohols, ketones and related esters; phenol and phenol derivatives; phenyl-substituted aliphatic alcohols and related aldehydes and esters; and sulfur-containing heterocyclic compounds). Specifications for the following food additives were revised: citric acid; gellan gum; polyoxyethylene (20) sorbitan monostearate; potassium aluminium silicate; and Quillaia extract (Type 2). Annexed to the report are tables summarizing the Committee's recommendations for dietary exposures to and toxicological evaluations of all of the food additives and flavouring agents considered at this meeting. PMID:26118220

  7. Climatic variability, plant phenology, and northern ungulates

    SciTech Connect

    Post, E.; Stenseth, N.C.

    1999-06-01

    Models of climate change predict that global temperatures and precipitation will increase within the next century, with the most pronounced changes occurring in northern latitudes and during winter. A large-scale atmospheric phenomenon, the North Atlantic Oscillation (NAO), is a strong determinant of both interannual variation and decadal trends in temperatures and precipitation during winter in northern latitudes, and its recent persistence in one extreme phase may be a substantial component of increases in global temperatures. Hence, the authors investigated the influences of large-scale climatic variability on plant phenology and ungulate population ecology by incorporating the NAO in statistical analyses of previously published data on: (1) the timing of flowering by plants in Norway, and (2) phenotypic and demographic variation in populations of northern ungulates. The authors analyzed 137 time series on plant phenology for 13 species of plants in Norway spanning up to 50 yr and 39 time series on phenotypic and demographic traits of 7 species of northern ungulates from 16 populations in North America and northern Europe spanning up to 30 yr.

  8. Seasonal distribution of zooplankton in the northern basin of Lake Chad

    USGS Publications Warehouse

    Robinson, A.H.; Robinson, Patricia K.

    1971-01-01

    More than 300 pairs of fine and coarse mesh plankton net samples were collected in the northern basin of Lake Chad during an 18-month period, June 1967 to November 1968. The seasonal distribution and abundance of the dominant species of Rotifera and Crustacea are given in addition to a general description of the hydrology and circulation of the northern basin of the lake. The composition and abundance of the zooplankton varied considerably over the sampling period; a generalized seasonal cycle is suggested. Synoptic estimates of absolute abundance are presented and compared to those in the southeastern portion of the lake.

  9. Manipulating crystallization with molecular additives.

    PubMed

    Shtukenberg, Alexander G; Lee, Stephanie S; Kahr, Bart; Ward, Michael D

    2014-01-01

    Given the importance of organic crystals in a wide range of industrial applications, the chemistry, biology, materials science, and chemical engineering communities have focused considerable attention on developing methods to control crystal structure, size, shape, and orientation. Tailored additives have been used to control crystallization to great effect, presumably by selectively binding to particular crystallographic surfaces and sites. However, substantial knowledge gaps still exist in the fundamental mechanisms that govern the formation and growth of organic crystals in both the absence and presence of additives. In this review, we highlight research discoveries that reveal the role of additives, either introduced by design or present adventitiously, on various stages of formation and growth of organic crystals, including nucleation, dislocation spiral growth mechanisms, growth inhibition, and nonclassical crystal morphologies. The insights from these investigations and others of their kind are likely to guide the development of innovative methods to manipulate crystallization for a wide range of materials and applications. PMID:24579880

  10. Additive Manufacturing of Hybrid Circuits

    NASA Astrophysics Data System (ADS)

    Sarobol, Pylin; Cook, Adam; Clem, Paul G.; Keicher, David; Hirschfeld, Deidre; Hall, Aaron C.; Bell, Nelson S.

    2016-07-01

    There is a rising interest in developing functional electronics using additively manufactured components. Considerations in materials selection and pathways to forming hybrid circuits and devices must demonstrate useful electronic function; must enable integration; and must complement the complex shape, low cost, high volume, and high functionality of structural but generally electronically passive additively manufactured components. This article reviews several emerging technologies being used in industry and research/development to provide integration advantages of fabricating multilayer hybrid circuits or devices. First, we review a maskless, noncontact, direct write (DW) technology that excels in the deposition of metallic colloid inks for electrical interconnects. Second, we review a complementary technology, aerosol deposition (AD), which excels in the deposition of metallic and ceramic powder as consolidated, thick conformal coatings and is additionally patternable through masking. Finally, we show examples of hybrid circuits/devices integrated beyond 2-D planes, using combinations of DW or AD processes and conventional, established processes.

  11. Tougher Addition Polyimides Containing Siloxane

    NASA Technical Reports Server (NTRS)

    St. Clair, T. L.; Maudgal, S.

    1986-01-01

    Laminates show increased impact resistances and other desirable mechanical properties. Bismaleamic acid extended by reaction of diaminosiloxane with maleic anhydride in 1:1 molar ratio, followed by reaction with half this molar ratio of aromatic dianhydride. Bismaleamic acid also extended by reaction of diaminosiloxane with maleic anhydride in 1:2 molar ratio, followed by reaction with half this molar ratio of aromatic diamine (Michael-addition reaction). Impact resistances improved over those of unmodified bismaleimide, showing significant increase in toughness. Aromatic addition polyimides developed as both matrix and adhesive resins for applications on future aircraft and spacecraft.

  12. Stratigraphy of the Martian northern plains

    NASA Technical Reports Server (NTRS)

    Tanaka, K. L.

    1993-01-01

    The northern plains of Mars are roughly defined as the large continuous region of lowlands that lies below Martian datum, plus higher areas within the region that were built up by volcanism, sedimentation, tectonism, and impacts. These northern lowlands span about 50 x 10(exp 6) km(sup 2) or 35 percent of the planet's surface. The age and origin of the lowlands continue to be debated by proponents of impact and tectonic explanations. Geologic mapping and topical studies indicate that volcanic, fluvial, and eolian deposition have played major roles in the infilling of this vast depression. Periglacial, glacial, fluvial, eolian, tectonic, and impact processes have locally modified the surface. Because of the northern plains' complex history of sedimentation and modification, much of their stratigraphy was obscured. Thus the stratigraphy developed is necessarily vague and provisional: it is based on various clues from within the lowlands as well as from highland areas within and bordering the plains. The results are summarized.

  13. Promoting Additive Acculturation in Schools.

    ERIC Educational Resources Information Center

    Gibson, Margaret A.

    1995-01-01

    A study focusing on 113 ninth graders of Mexican descent indicates that most students and their parents adhere to a strategy of additive acculturation (incorporating skills of the new culture and language), but that the school curriculum and general school climate devalue Mexican culture. (SLD)

  14. Individualized Additional Instruction for Calculus

    ERIC Educational Resources Information Center

    Takata, Ken

    2010-01-01

    College students enrolling in the calculus sequence have a wide variance in their preparation and abilities, yet they are usually taught from the same lecture. We describe another pedagogical model of Individualized Additional Instruction (IAI) that assesses each student frequently and prescribes further instruction and homework based on the…

  15. Out of bounds additive manufacturing

    SciTech Connect

    Holshouser, Chris; Newell, Clint; Palas, Sid; Love, Lonnie J.; Kunc, Vlastimil; Lind, Randall F.; Lloyd, Peter D.; Rowe, John C.; Blue, Craig A.; Duty, Chad E.; Peter, William H.; Dehoff, Ryan R.

    2013-03-01

    Lockheed Martin and Oak Ridge National Laboratory are working on an additive manufacturing system capable of manufacturing components measured not in terms of inches or feet, but multiple yards in all dimensions with the potential to manufacture parts that are completely unbounded in size.

  16. The Additive Property of Energy.

    ERIC Educational Resources Information Center

    Tsaoussis, Dimitris S.

    1995-01-01

    Presents exercises that analyze the additive property of energy. Concludes that if a body has more than one component of energy depending on the same physical quantity, the body's total energy will be the algebraic sum of the components if a linear relationship exists between the energy components and that physical quantity. (JRH)

  17. Tinkertoy Color-Addition Device.

    ERIC Educational Resources Information Center

    Ferguson, Joe L.

    1995-01-01

    Describes construction and use of a simple home-built device, using an overhead projector, for use in demonstrations of the addition of various combinations of red, green, and blue light. Useful in connection with discussions of color, color vision, or color television. (JRH)

  18. Silage Additives and Management Issues

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Inoculants are the most common silage additives in the United States. These products contain lactic acid bacteria to supplement the lactic acid bacteria naturally on the crop and help insure a consistent fermentation in the silo. There are three types of inoculants: homofermentative lactic acid bact...

  19. Tetrasulfide extreme pressure lubricant additives

    SciTech Connect

    Gast, L.E.; Kenney, H.E.; Schwab, A.W.

    1980-08-19

    A novel class of compounds has been prepared comprising the tetrasulfides of /sup 18/C hydrocarbons, /sup 18/C fatty acids, and /sup 18/C fatty and alkyl and triglyceride esters. These tetrasulfides are useful as extreme pressure lubricant additives and show potential as replacements for sulfurized sperm whale oil.

  20. Evaluation of certain food additives.

    PubMed

    2012-01-01

    This report represents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of various food additives, including flavouring agents, with a view to concluding as to safety concerns and to preparing specifications for identity and purity. The first part of the report contains a general discussion of the principles governing the toxicological evaluation of and assessment of dietary exposure to food additives, including flavouring agents. A summary follows of the Committee's evaluations of technical, toxicological and dietary exposure data for five food additives (magnesium dihydrogen diphosphate; mineral oil (medium and low viscosity) classes II and III; 3-phytase from Aspergillus niger expressed in Aspergillus niger; serine protease (chymotrypsin) from Nocardiopsis prasina expressed in Bacillus licheniformis; and serine protease (trypsin) from Fusarium oxysporum expressed in Fusarium venenatum) and 16 groups of flavouring agents (aliphatic and aromatic amines and amides; aliphatic and aromatic ethers; aliphatic hydrocarbons, alcohols, aldehydes, ketones, carboxylic acids and related esters, sulfides, disulfides and ethers containing furan substitution; aliphatic linear alpha,beta-unsaturated aldehydes, acids and related alcohols, acetals and esters; amino acids and related substances; epoxides; furfuryl alcohol and related substances; linear and branched-chain aliphatic, unsaturated, unconjugated alcohols, aldehydes, acids and related esters; miscellaneous nitrogen-containing substances; phenol and phenol derivatives; pyrazine derivatives; pyridine, pyrrole and quinoline derivatives; saturated aliphatic acyclic branched-chain primary alcohols, aldehydes and acids; simple aliphatic and aromatic sulfides and thiols; sulfur-containing heterocyclic compounds; and sulfur-substituted furan derivatives). Specifications for the following food additives were revised: ethyl cellulose, mineral oil (medium viscosity), modified starches and titanium

  1. Climate impacts on northern Canada: regional background.

    PubMed

    Prowse, Terry D; Furgal, Chris; Bonsal, Barrie R; Peters, Daniel L

    2009-07-01

    Understanding the implications of climate change on northern Canada requires a background about the size and diversity of its human and biogeophysical systems. Occupying an area of almost 40% of Canada, with one-third of this contained in Arctic islands, Canada's northern territories consist of a diversity of physical environments unrivaled around the circumpolar north. Major ecozones composed of a range of landforms, climate, vegetation, and wildlife include: Arctic, boreal and taiga cordillera; boreal and taiga plains; taiga shield; and northern and southern Arctic. Although generally characterized by a cold climate, there is an enormous range in air temperature with mean annual values being as high as -5 degrees C in the south to as low as -20 degrees C in the high Arctic islands. A similar contrast characterizes precipitation, which can be > 700 mm y(-1) in some southern alpine regions to as low as 50 mm y(-1) over islands of the high Arctic. Major freshwater resources are found within most northern ecozones, varying from large glaciers or ice caps and lakes to extensive wetlands and peat lands. Most of the North's renewable water, however, is found within its major river networks and originates in more southerly headwaters. Ice covers characterize the freshwater systems for multiple months of the year while permafrost prevails in various forms, dominating the terrestrial landscape. The marine environment, which envelops the Canadian Arctic Archipelago, is dominated by seasonal to multiyear sea ice often several meters thick that plays a key role in the regional climate. Almost two-thirds of northern Canadian communities are located along coastlines with the entire population being just over 100 000. Most recent population growth has been dominated by an expansion of nonaboriginals, primarily the result of resource development and the growth of public administration. The economies of northern communities, however, remain quite mixed with traditional land

  2. ABCGheritage project - promoting geotourism in northern Finland, northern Norway and the Kola Peninsula

    NASA Astrophysics Data System (ADS)

    Pihlaja, Jouni; Johansson, Peter; Lauri, Laura S.

    2014-05-01

    Nature tourism has been a growing business sector in the Barents area during the recent decades. With the purpose to develop nature tourism in a sustainable way, a cooperation project ABCGheritage - Arctic Biological, Cultural and Geological Heritage has been carried out. Project has received partial funding from the EU Kolarctic ENPI program. In the geoheritage part of the project the main activities were aimed to develop pro-environmental ways of geotourism in the area. The three main participants in the geoheritage part of the project are the Geological Survey of Finland, Northern Finland Office, the Geological Institute of the Kola Science Centre of the Russian Academy of Sciences and Bioforsk Soil and Environment from northeastern Norway. The duration of the project is 2012-2014 and most of the work has already been completed even if most of the results are not published yet. Totally ten different tasks have been implemented in the geological part of the project. The largest task has been the preparation of a geological outdoor map and guide book of the Khibiny Tundra locating in the central part of the Kola Peninsula. In Finland already 11 such maps have been published, and the experiences gained during their production have been used in this project, too. Geological heritage trails to the Khibiny Tundra have also been created and they will be drawn on the map. The second concrete result is the Barents Tour for Geotourist -guide, which will be published as a guide book, web pages and an exhibition. The route comprises ca 35 best geological demonstration sites along the circle route from northern Finland to northeastern Norway, from there to Kola Peninsula and then back to Finland. Information of the route will be available for all interested travelers. In addition to the geological outdoor map of the Khibiny Tundra and "Barents Tour for Geotourists"-guide, the primary outputs of the project are the geological nature trails on the field, geological

  3. Rural telemedicine project in northern New Mexico

    SciTech Connect

    Zink, S.; Hahn, H.; Rudnick, J.; Snell, J.; Forslund, D.; Martinez, P.

    1998-12-31

    A virtual electronic medical record system is being deployed over the Internet with security in northern New Mexico using TeleMed, a multimedia medical records management system that uses CORBA-based client-server technology and distributed database architecture. The goal of the NNM Rural Telemedicine Project is to implement TeleMed into fifteen rural clinics and two hospitals within a 25,000 square mile area of northern New Mexico. Evaluation of the project consists of three components: job task analysis, audit of immunized children, and time motion studies. Preliminary results of the evaluation components are presented.

  4. Coastal geomorphology of the Martian northern plains

    NASA Technical Reports Server (NTRS)

    Parker, Timothy J.; Gorsline, Donn S.; Saunders, Stephen R.; Pieri, David C.; Schneeberger, Dale M.

    1993-01-01

    The paper considers the question of the formation of the outflow channels and valley networks discovered on the Martian northern plains during the Mariner 9 mission. Parker and Saunders (1987) and Parker et al. (1987, 1989) data are used to describe key features common both in the lower reaches of the outflow channels and within and along the margins of the entire northern plains. It is suggested, that of the geological processes capable of producing similar morphologies on earth, lacustrine or marine deposition and subsequent periglacial modification offer the simplest and most consistent explanation for the suit of features found on Mars.

  5. Geoenvironmental Investigations of the Humboldt River Basin, Northern Nevada

    USGS Publications Warehouse

    Stillings, Lisa L.

    2003-01-01

    Northern Nevada is one of the world's foremost regions of gold production. The Humboldt River Basin (HRB) covers 43,500 km2 in northern Nevada (Crompton, 1995), and it is home to approximately 18 active gold and silver mines (Driesner and Coyner, 2001) among at least 55 significant metallic mineral deposits (Long and others, 1998). Many of the gold mines are along the Carlin trend in the east-central portion of the HRB, and together they have produced 50 million ounces of gold from 1962 (when the Carlin mine first opened) through April 2002 (Nevada Mining Association, 2002). Mining is not new to the region, however. Beginning in 1849, mining has taken place in numerous districts that cover 39 percent of the land area in the HRB (Tingley, 1998). In addition to gold and silver, As, Ba, Cu, Fe, Hg, Li, Mn, Mo, Pb, S, Sb, V, W, Zn, and industrial commodities such as barite, limestone, fluorite, sand and gravel, gypsum, gemstones, pumice, zeolites, and building stone, have been extracted from the HRB (McFaul and others, 2000). All papers within this series of investigations can be found as lettered chapters of USGS Bulletin 2210, Geoenvironmental Investigations of the Humboldt River Basin, Northern Nevada. Each chapter is available separately online. The data and software utilized in this product (Chapter F) permit the user to view and analyze the geographic relationships among chemistry of stream sediments and surface waters, geology, and various cartographic base information such as but not limited to cities, county boundaries, and land ownership. Data for this product were compiled and or produced as part of a mineral and environmental assessment of the Humboldt River basin conducted by the U.S. Geological Survey between 1995 - 2000.

  6. Development of a dot blot assay with antibodies to recombinant “core” 14-3-3 protein: Evaluation of its usefulness in diagnosis of Creutzfeldt–Jakob disease

    PubMed Central

    Subramanian, Sarada; Mahadevan, Anita; Satishchandra, Parthasarathy; Shankar, Susarla Krishna

    2016-01-01

    Background and Purpose: Definitive diagnosis of Creutzfeldt–Jakob disease (CJD) requires demonstration of infective prion protein (PrPSc) in brain tissues by immunohistochemistry or immunoblot, making antemortem diagnosis of CJD difficult. The World Health Organization (WHO) recommends detection of 14-3-3 protein in cerebrospinal fluid (CSF) in cases of dementia, with clinical correlation, as a useful diagnostic marker for CJD, obviating the need for brain biopsy. This facility is currently available in only a few specialized centers in the West and no commercial kit is available for clinical diagnostic use in India. Hence the objective of this study was to develop an in-house sensitive assay for quantitation of 14-3-3 protein and to evaluate its diagnostic potential to detect 14-3-3 proteins in CSF as a biomarker in suspected cases of CJD. Materials and Methods: A minigene expressing the “core” 14-3-3 protein was synthesized by overlapping polymerase chain reaction (PCR) and the recombinant protein was produced by employing a bacterial expression system. Polyclonal antibodies raised in rabbit against the purified recombinant protein were used for developing a dot blot assay with avidin-biotin technology for signal amplification and quantitation of 14-3-3 protein in CSF. Results: The results in the present study suggest the diagnostic potential of the dot blot method with about 10-fold difference (P< 0.001) in the CSF levels of 14-3-3 protein between the CJD cases (N= 50) and disease controls (N= 70). The receiver operating characteristic (ROC) analysis of the results suggested an optimal cutoff value of 2 ng/mL. Conclusions: We have developed an indigenous, economical, and sensitive dot blot method for the quantitation of 14-3-3 protein in CSF. PMID:27293331

  7. The Influence of European Pollution on Ozone in the Near East and Northern Africa

    NASA Technical Reports Server (NTRS)

    Duncan, B. N.; West, J. J.; Yoshida, Y.; Fiore, A. M.; Ziemke, J. R.

    2008-01-01

    We present a modeling study of the long-range transport of pollution from Europe, showing that European emissions regularly elevate surface ozone by as much as 20 ppbv in summer in northern Africa and the Near East. European emissions cause 50-150 additional violations per year (i.e. above those that would occur without European pollution) of the European health standard for ozone (8-h average greater than 120 micrograms per cubic meters or approximately 60 ppbv) in northern Africa and the Near East. We estimate that European ozone pollution is responsible for 50 000 premature mortalities globally each year, of which the majority occurs outside of Europe itself, including 37% (19 000) in northern Africa and the Near East. Much of the pollution from Europe is exported southward at low altitudes in summer to the Mediterranean Sea, northern Africa and the Near East, regions with favorable photochemical environments for ozone production. Our results suggest that assessments of the human health benefits of reducing ozone precursor emissions in Europe should include effects outside of Europe, and that comprehensive planning to improve air quality in northern Africa and the Near East likely needs to address European emissions.

  8. Genetics Show Current Decline and Pleistocene Expansion in Northern Spotted Owls

    USGS Publications Warehouse

    Funk, W. Chris; Forsman, Eric D.; Mullins, Thomas D.; Haig, Susan M.

    2008-01-01

    The northern spotted owl (Strix occidentalis caurina) is one of the most controversial threatened subspecies ever listed under the U.S. Endangered Species Act. Because of concern for persistence of the subspecies, logging on Federal lands in the U.S. Pacific Northwest was dramatically reduced under the Northwest Forest Plan in 1994. Despite protection of its remaining forest habitat, recent field studies show continued demographic declines of northern spotted owls. One potential threat to northern spotted owls that has not yet been shown is loss of genetic variation from population bottlenecks that can increase inbreeding depression and decrease adaptive potential. Here, we show recent genetic bottlenecks in northern spotted owls using a large genetic dataset (352 individuals from across the subspecies' range and 11 microsatellite loci). The signature of bottlenecks was strongest in Washington State, in agreement with field data. Interestingly, we also found a genetic signature of Pleistocene expansion in the same study areas where recent bottlenecks were shown. Our results provide independent evidence that northern spotted owls have recently declined, and suggest that loss of genetic variation is an emerging threat to the subspecies' persistence. Reduced effective population size (Ne), shown here in addition to field evidence for demographic decline, highlights the increasing vulnerability of this bird to extinction.

  9. Decontamination formulation with sorbent additive

    DOEpatents

    Tucker; Mark D. , Comstock; Robert H.

    2007-10-16

    A decontamination formulation and method of making that neutralizes the adverse health effects of both chemical and biological compounds, especially chemical warfare (CW) and biological warfare (BW) agents, and toxic industrial chemicals. The formulation provides solubilizing compounds that serve to effectively render the chemical and biological compounds, particularly CW and BW compounds, susceptible to attack, and at least one reactive compound that serves to attack (and detoxify or kill) the compound. The formulation includes at least one solubilizing agent, a reactive compound, a bleaching activator, a sorbent additive, and water. The highly adsorbent, water-soluble sorbent additive (e.g., sorbitol or mannitol) is used to "dry out" one or more liquid ingredients, such as the liquid bleaching activator (e.g., propylene glycol diacetate or glycerol diacetate) and convert the activator into a dry, free-flowing powder that has an extended shelf life, and is more convenient to handle and mix in the field.

  10. Robust stability under additive perturbations

    NASA Technical Reports Server (NTRS)

    Bhaya, A.; Desoer, C. A.

    1985-01-01

    A MIMO linear time-invariant feedback system 1S(P,C) is considered which is assumed to be U-stable. The plant P is subjected to an additive perturbation Delta P which is proper but not necessarily stable. It is proved that the perturbed system is U-stable if and only if Delta P(I + Q x Delta P) exp -1 is U-stable.

  11. Additive manufacturing of hybrid circuits

    DOE PAGESBeta

    Bell, Nelson S.; Sarobol, Pylin; Cook, Adam; Clem, Paul G.; Keicher, David M.; Hirschfeld, Deidre; Hall, Aaron Christopher

    2016-03-26

    There is a rising interest in developing functional electronics using additively manufactured components. Considerations in materials selection and pathways to forming hybrid circuits and devices must demonstrate useful electronic function; must enable integration; and must complement the complex shape, low cost, high volume, and high functionality of structural but generally electronically passive additively manufactured components. This article reviews several emerging technologies being used in industry and research/development to provide integration advantages of fabricating multilayer hybrid circuits or devices. First, we review a maskless, noncontact, direct write (DW) technology that excels in the deposition of metallic colloid inks for electrical interconnects.more » Second, we review a complementary technology, aerosol deposition (AD), which excels in the deposition of metallic and ceramic powder as consolidated, thick conformal coatings and is additionally patternable through masking. As a result, we show examples of hybrid circuits/devices integrated beyond 2-D planes, using combinations of DW or AD processes and conventional, established processes.« less

  12. Hydrocarbon Induced Seismicity in Northern Netherlands

    NASA Astrophysics Data System (ADS)

    Dost, B.; Spetzler, J.; Kraaijpoel, D.; Caccavale, M.

    2015-12-01

    The northern Netherlands has been regarded aseismic until the first earthquakes started in 1986, after more than 25 years of gas production from the one of the largest on-shore gas-fields in the World, the Groningen field, and accompanying smaller gas fields. Due to the shallow sources, at approximately 3 km depth, even small magnitude events cause considerable damage to buildings in the region. Since the largest recorded event in the Groningen field in 2012 with ML= 3,6, more than 30.000 damage claims were received by the mining company. Since 1995 a seismic monitoring network is operational in the region, consisting of 8 200m deep boreholes with 4 levels of 3C 4,5 Hz geophones. The network was designed for a location threshold of ML=1,5 over a 40x 80 km region. Average station separation was 20 km. At the end of 2014, 245 events have been recorded with ML ≥ 1,5, out of a total of 1100. Since 2003 a new mining law is in place in the Netherlands, which requires for each gas field in production a seismic risk analysis. Initially, due to the small number of events for specific fields, a general hazard (PSHA) was calculated for all gas-fields and a maximum magnitude was estimated at ML = 3,9. Since 2003 an increase in the activity rate is observed for the Groningen field, leading to the development of new models and a re-assessment of parameters like the maximum magnitude. More recently these models are extended to seismic risk, where also the fragility of the regional buildings is taken into account. Understanding the earthquake process is essential in taking mitigation measures. Continued research is focused on reducing the uncertainties in the hazard and risk models and is accompanied by an upgrade of the monitoring network. In 2014 a new dense network was designed to monitor the Groningen gas field in this region (30*40 km) with an average separation of 4 km. This allows an improved location threshold (M>0,5) and location accuracy (50-100m). A detailed P- and S

  13. Lithostratigraphical analysis in northern Sinus Meridiani, Mars.

    NASA Astrophysics Data System (ADS)

    Baioni, D.; Sgavetti, M.; Tampella, G.; Wezel, F. C.

    2012-04-01

    In the Terra Meridiani- Arabia Terra region of Mars, layered rock units with a range of stratal patterns have been widely reported in the literature, and the record of the spectral signatures indicative of hydrate minerals suggests the existence of aqueous activity in at least some of the stratigraphic units. . In this work we investigated the area in the northern region of Sinus Meridiani located between 2°N to 3°N latitude and 2°W to 1°E longitude, using multiple remote sensing data sets to identify, characterize in detail, and understand the origin and evolution of the selected units in this region of Mars. MOLA and HiRISE data were used in combination to classify and correlate surface units recorded in distinct stratigraphic logs, based on planimetric configuration, topography, morphological and litho-facies characteristics. In particular, detailed analyses were focused on the layered units discovered in the walls of two impact craters located at a distance of about 100 km to each other within the study area. The stratigraphic interval analysed here corresponds to the upper part of the well known "light-toned layered deposits" identified by several authors. Distinct competent layers are observed in the internal slopes of both craters. Our observations indicate that the lithostratigraphic sections can be divided in three units. The lower unit consists of very thick bodies of light-toned bouldery breccia deposits. In the HiRISE images these white-coloured beds are composed of irregularly-shaped, white nodules which are either isolated or coalescent, distributed between bedding planes. The middle unit mainly consists of alternating thin strong layers and thicker sequences of relatively weak layers, horizontally bedded. The strong, competent layers maintain steeper erosional profiles and play a major role in controlling the overall shape and geomorphology of the wall slopes. The upper unit can be traced laterally in the surrounding ground level, displaying mesa

  14. Subduction zone guided waves in Northern Chile

    NASA Astrophysics Data System (ADS)

    Garth, Thomas; Rietbrock, Andreas

    2016-04-01

    Guided wave dispersion is observed in subduction zones as high frequency energy is retained and delayed by low velocity structure in the subducting slab, while lower frequency energy is able to travel at the faster velocities associated with the surrounding mantle material. As subduction zone guided waves spend longer interacting with the low velocity structure of the slab than any other seismic phase, they have a unique capability to resolve these low velocity structures. In Northern Chile, guided wave arrivals are clearly observed on two stations in the Chilean fore-arc on permanent stations of the IPOC network. High frequency (> 5 Hz) P-wave arrivals are delayed by approximately 2 seconds compared to the low frequency (< 2 Hz) P-wave arrivals. Full waveform finite difference modelling is used to test the low velocity slab structure that cause this P-wave dispersion. The synthetic waveforms produced by these models are compared to the recorded waveforms. Spectrograms are used to compare the relative arrival times of different frequencies, while the velocity spectra is used to constrain the relative amplitude of the arrivals. Constraining the waveform in these two ways means that the full waveform is also matched, and the low pass filtered observed and synthetic waveforms can be compared. A combined misfit between synthetic and observed waveforms is then calculated following Garth & Rietbrock (2014). Based on this misfit criterion we constrain the velocity model by using a grid search approach. Modelling the guided wave arrivals suggest that the observed dispersion cannot be solely accounted for by a single low velocity layer as suggested by previous guided wave studies. Including dipping low velocity normal fault structures in the synthetic model not only accounts for the observed strong P-wave coda, but also produces a clear first motion dispersion. We therefore propose that the lithospheric mantle of the subducting Nazca plate is highly hydrated at intermediate

  15. Evaluation of certain food additives.

    PubMed

    2009-01-01

    This report represents the conclusions of a Joint FAO/WHO Expert Committee convened to evaluate the safety of various food additives, including flavouring agents, with a view to recommending acceptable daily intakes (ADIs) and to preparing specifications for identity and purity. The first part of the report contains a general discussion of the principles governing the toxicological evaluation and assessment of intake of food additives (in particular, flavouring agents). A summary follows of the Committee's evaluations of technical, toxicological and intake data for certain food additives (asparaginase from Aspergillus niger expressed in A. niger, calcium lignosulfonate (40-65), ethyl lauroyl arginate, paprika extract, phospholipase C expressed in Pichia pastoris, phytosterols, phytostanols and their esters, polydimethylsiloxane, steviol glycosides and sulfites [assessment of dietary exposure]) and 10 groups of related flavouring agents (aliphatic branched-chain saturated and unsaturated alcohols, aldehydes, acids and related esters; aliphatic linear alpha,beta-unsaturated aldehydes, acids and related alcohols, acetals and esters; aliphatic secondary alcohols, ketones and related esters; alkoxy-substituted allylbenzenes present in foods and essential oils and used as flavouring agents; esters of aliphatic acyclic primary alcohols with aliphatic linear saturated carboxylic acids; furan-substituted aliphatic hydrocarbons, alcohols, aldehydes, ketones, carboxylic acids and related esters, sulfides, disulfides and ethers; miscellaneous nitrogen-containing substances; monocyclic and bicyclic secondary alcohols, ketones and related esters; hydroxy- and alkoxy-substituted benzyl derivatives; and substances structurally related to menthol). Specifications for the following food additives were revised: canthaxanthin; carob bean gum and carob bean gum (clarified); chlorophyllin copper complexes, sodium and potassium salts; Fast Green FCF; guar gum and guar gum (clarified

  16. Fire-Retardant Polymeric Additives

    NASA Technical Reports Server (NTRS)

    Williams, Martha K.; Smith, Trent M.

    2011-01-01

    Polyhydroxyamide (PHA) and polymethoxyamide (PMeOA) are fire-retardant (FR) thermoplastic polymers and have been found to be useful as an additive for imparting fire retardant properties to other compatible, thermoplastic polymers (including some elastomers). Examples of compatible flammable polymers include nylons, polyesters, and acrylics. Unlike most prior additives, PHA and PMeOA do not appreciably degrade the mechanical properties of the matrix polymer; indeed, in some cases, mechanical properties are enhanced. Also, unlike some prior additives, PHA and PMeOA do not decompose into large amounts of corrosive or toxic compounds during combustion and can be processed at elevated temperatures. PMeOA derivative formulations were synthesized and used as an FR additive in the fabrication of polyamide (PA) and polystyrene (PS) composites with notable reduction (>30 percent for PS) in peak heat release rates compared to the neat polymer as measured by a Cone Calorimeter (ASTM E1354). Synergistic effects were noted with nanosilica composites. These nanosilica composites had more than 50-percent reduction in peak heat release rates. In a typical application, a flammable thermoplastic, thermoplastic blend, or elastomer that one seeks to render flame-retardant is first dry-mixed with PHA or PMeOA or derivative thereof. The proportion of PHA or PMeOA or derivative in the mixture is typically chosen to lie between 1 and 20 weight percent. The dry blend can then be melt-extruded. The extruded polymer blend can further be extruded and/or molded into fibers, pipes, or any other of a variety of objects that may be required to be fire-retardant. The physical and chemical mechanisms which impart flame retardancy of the additive include inhibiting free-radical oxidation in the vapor phase, preventing vaporization of fuel (the polymer), and cooling through the formation of chemical bonds in either the vapor or the condensed phase. Under thermal stress, the cyclic hydroxyl/ methoxy

  17. Complete Alpine reworking of the northern Menderes Massif, western Turkey

    NASA Astrophysics Data System (ADS)

    Cenki-Tok, B.; Expert, M.; Işık, V.; Candan, O.; Monié, P.; Bruguier, O.

    2016-07-01

    This study focuses on the petrology, geochronology and thermochronology of metamorphic rocks within the northern Menderes Massif in western Turkey. Metasediments belonging to the cover series of the Massif record pervasive amphibolite-facies metamorphism culminating at ca. 625-670 °C and 7-9 kbars. U-Th-Pb in situ ages on monazite and allanite from these metapelites record crustal thickening and nappe stacking associated with the internal imbrication of the Anatolide-Taurides platform during the Eocene. In addition, new 39Ar/40Ar single muscovite grain analyses on deformed rocks were performed in three localities within the northern Menderes Massif and ages range from 19.8 to 25.5 Ma. These mylonites may be related to both well-known detachments, Simav to the north and Alaşehir to the south, which accommodate Oligo-Miocene exhumation of the Menderes core complex. U-Th-Pb data on monazite grains (22.2 ± 0.2 Ma) from migmatites emplaced within the Simav detachment confirm these ages.

  18. Potential cretaceous play in the Rharb basin of northern Morocco

    SciTech Connect

    Jobidon, G.P. )

    1993-09-01

    The autochthonous Cretaceous in the Rharb basin of northern Morocco is located underneath a cover of neogene sediments and of the Prerif nappe olistostrome, which was emplaced during the Tortonian 7 m.y. The presence of infranappe Cretaceous sediments is documented in a few onshore wells in the Rharb basin and in the adjacent Prerif Rides area, as well as in the Rif Mountains. Their presence in the deeper portion of the Rharb basin is difficult to detail because of poor seismic resolution data beneath dispersive prerif nappe. A recent study of offshore seismic data acquired by PCIAC in 1987 indicates that the infranappe interval can be more than 1500 m thick in some of the offshore Kenitra area. These sediments have seismic signatures that would correspond to Middle Cretaceous transgressions, culminating with a Turonian highstand. Their deposition systems were located on the northern and western flanks of the Meseta and were followed by a hiatus lasting until the Miocene. Regional studies of gravity and magnetic data provide and additional understanding of the Rif province, its evolution, and the possible presence of autochthonous Cretaceous sediments below the prerif nappe cover. The infranappe of Rharb basin has a good potential to develop into a major hydrocarbon play with the presence of middle Cretaceous reservoir rocks, Turonian-Cenomanian black shale source rocks, as well as the timely combination of trap formation, source rock maturation, and hydrocarbon migration.

  19. Sexual behavior and attitudes of university students in Northern Ireland.

    PubMed

    Sneddon, I; Kremer, J

    1992-06-01

    Two surveys of a Northern Ireland student sample were conducted in 1987 and 1988. A total of 419 female and 201 male subjects completed self-administered anonymous questionnaires concerning their behavior, knowledge, and attitudes towards sex, AIDS, homosexuality, contraception, and relationships. Results indicated a relatively low level of sexual experience, and for those with experience, relatively few partners. The possible influences of gender and religiosity on sexual behavior and attitudes, in the context of Northern Ireland, are discussed. Subjects reported considerable variation in the amount of sex education, but the majority received little or none. This student sample held relatively conservative attitudes towards love, sex, and marriage and this was particularly true for females and for regular churchgoers. In addition, attitudes towards homosexuality were negative (particularly among regular churchgoers). Attitudes towards contraception were more positive than expected among Catholic subjects, and few indicated that they would refuse to use contraceptives on principle. Responses to items about AIDS were highly uniform, suggesting that much of the information made available to the public has been absorbed. However, the lack of uniformity of response to more general items about sex, relationships, and contraception may indicate that fundamental changes in sexual behavior are unlikely to be brought about by influencing a rather narrowly defined set of attitudes about AIDS. PMID:1610290

  20. Tracking rainfall in the northern Mediterranean borderlands during sapropel deposition

    NASA Astrophysics Data System (ADS)

    Toucanne, Samuel; Angue Minto'o, Charlie Morelle; Fontanier, Christophe; Bassetti, Maria-Angela; Jorry, Stephan J.; Jouet, Gwenaël

    2015-12-01

    The role of mid-latitude precipitation in the hydrological forcing leading to the deposition of sapropels in the Mediterranean Sea remains unclear. The new GDEC-4-2 borehole, East Corsica margin (northern Tyrrhenian Sea), provides the first precisely dated evidence for enhanced rainfall in the Western Mediterranean during warm intervals of interglacial periods over the last 547 kyr. Comparison of GDEC-4-2 proxy records with pollen sequences and speleothems from the central and eastern Mediterranean reveals that these pluvial events were regional in character and occurred probably in response to the intensification of the Mediterranean storm track along the northern Mediterranean borderlands in autumn/winter. Our dataset suggests that the timing of maxima of the Mediterranean autumn/winter storm track precipitation coincide with that of the North African summer monsoon and sapropel deposition. Besides highlighting a close coupling between mid- and low-latitude hydrological changes, our findings suggest that during warm intervals of interglacial periods the reduced sea-surface water salinities, together with the high flux of nutrient and organic matter, produced by the monsoonal Nile (and wadi-systems) floods, were maintained throughout the winter by the Mediterranean rainfall. This provides an important additional constraint on the hydrological perturbation causing sapropel formation.

  1. Jurassic extension and Alpine inversion of the northern Morocco

    SciTech Connect

    Zizi, M. )

    1993-09-01

    The lower Mesozoic half grabens of northern Morocco form part of an extensional system that is related to the opening of the western Tethys. They appear to be somewhat younger than the Triassic-Jurassic systems associated with the opening the Atlantic Ocean. During the Tertiary and as consequence of the Alpine collision of Africa with Europe, these half graben systems were inverted as shown by the High and the Middle Atlas mountains. Seismic illustrations of similar but smaller inversion structures are available from the Guercif area and the [open quotes]Rides Prerifaines[close quotes] of northern Morocco. These seismic profiles serve as small models for the much larger Atlas Mountains. In the Guercif area, the inversions are limited in scope, but in the [open quotes]Ride Prerifaines[close quotes] are extensive decollement systems that sole out in the Triassic evaporites. These systems evolve into complex thrust faults and associated lateral ramps that are strongly influenced by the configuration of the Jurassic transtensional systems. Significant hydrocarbon accumulation have been known for some time from the [open quotes]Rides Prerifaines.[close quotes] A review of the geometry of the inverted half-graben systems, combined with detailed stratigraphic studies, is likely to lead to the discovery of additional reserves in the area.

  2. Carbon, nitrogen, and phosphorus mineralization in northern wetlands

    SciTech Connect

    Bridgham, S.D.; Updegraff, K.; Pastor, J.

    1998-07-01

    The authors examined rates of C, N, and P mineralization in soils from 16 northern Minnesota wetlands that occur across an ombrotrophic-minerotrophic gradient. Soils were incubated at 30 C under aerobic and anaerobic conditions for 59 wk, and the results were fit with a two-pool kinetic model. Additionally, 39 different soil quality variables were used in a principal components analysis (PCA) to predict mineralization rates. Mineralization of C, N, and P differed significantly among wetland types, aeration status (aerobic vs. anaerobic), and their interaction term. Despite low total soil N and P, there was a rapid turnover of the nutrient pools in ombrotrophic sites, particularly under aerobic conditions. On a volumetric basis, C and N mineralization increased in a predictable manner across the ombrotrophic-minerotrophic gradient, largely due to increasing soil bulk density. However, P mineralization per cubic centimeter remained relatively high in the bogs. The higher total P content of more minerotrophic soils appears to be offset by greater P immobilization due to geochemical sorption, yielding overall lower availability. Large differences in mineralization rates in northern wetland communities demonstrate that climate change models should not consider these areas as homogeneous entities.

  3. Landscape Features Shape Genetic Structure in Threatened Northern Spotted Owls

    USGS Publications Warehouse

    Funk, W. Chris; Forsman, Eric D.; Mullins, Thomas D.; Haig, Susan M.

    2008-01-01

    Several recent studies have shown that landscape features can strongly affect spatial patterns of gene flow and genetic variation. Understanding landscape effects on genetic variation is important in conservation for defining management units and understanding movement patterns. The landscape may have little effect on gene flow, however, in highly mobile species such as birds. We tested for genetic breaks associated with landscape features in the northern spotted owl (Strix occidentalis caurina), a threatened subspecies associated with old forests in the U.S. Pacific Northwest and extreme southwestern Canada. We found little evidence for distinct genetic breaks in northern spotted owls using a large microsatellite dataset (352 individuals from across the subspecies' range genotyped at 10 loci). Nonetheless, dry low-elevation valleys and the Cascade and Olympic Mountains restrict gene flow, while the Oregon Coast Range facilitates it. The wide Columbia River is not a barrier to gene flow. In addition, inter-individual genetic distance and latitude were negatively related, likely reflecting northward colonization following Pleistocene glacial recession. Our study shows that landscape features may play an important role in shaping patterns of genetic variation in highly vagile taxa such as birds.

  4. Carbon dioxide emissions from estuaries of northern and northeastern Brazil

    PubMed Central

    Noriega, Carlos; Araujo, Moacyr

    2014-01-01

    The carbon dioxide flux through the air–water interface of coastal estuarine systems must be quantified to understand the regional balance of carbon and its transport through adjacent coastal regions. We estimated and calculated the emissions of carbon dioxide (FCO2) and the partial pressure of CO2 (pCO2) values in 28 estuarine environments at a variety of spatial scales in the northern and northeastern regions of Brazil. The results showed a mean FCO2 (water to air) of 55 ± 45 mmol·m−2·d−1. Additionally, a negative correlation between dissolved oxygen saturation and pCO2 was observed, indicating a control by biological processes and especially by organic matter degradation. This leads to increased dissolved CO2 concentration in estuarine waters which results in a pCO2 that reached 8,638 μatm. Our study suggests that northern and northeastern Brazilian estuaries act as sources of atmospheric CO2. The range of pCO2 observed were similar to those found in inner estuaries in other places around the world, with the exception of a few semi-arid estuaries (Köppen climate classification – BSh) in which record low levels of pCO2 have been detected. PMID:25145418

  5. Palaeoindian occupation of the Atacama Desert, northern Chile

    NASA Astrophysics Data System (ADS)

    Grosjean, Martin; Núñez, Lautaro; Cartajena, Isabel

    2005-10-01

    Palaeoindian occupation of the Atacama Desert in northern Chile has been found between 12 600 and 10 200 cal. yr BP. The new site at Salar Punta Negra (24°28S/60°53W/2976 m) includes about 1000 classifiable, mostly unifacial artefacts and, uniquely, three different diagnostic types of early projectile points. Two of the Lateglacial/early Holocene projectile types have wide distribution and are known from different geographical areas in South America: the Palaeoindian Fell fish-tail point mainly from the southern cone of South America, and the triangular Tuina points typical of the Puna of the south-central Andes in northern Chile and northwestern Argentina. In addition, we found a third type, a stemmed point typical for the Salar Punta Negra. Filling a large geographical gap of Fell occupation, the site at Salar Punta Negra provides evidence for generally much higher mobility and diversity of early cultures, and supports an Andean-Pacific route for early human exploration of South America to the south through the desert at intermediate altitudes. Contemporaneous high-amplitude climatic changes were fundamental preconditions to provide adequate environments and habitats, and to make Palaeoindian hunting-gathering occupation possible in the Atacama Desert.

  6. Thermohaline finestructure observed near the northern Philippine coast

    NASA Astrophysics Data System (ADS)

    Wang, Fan; Li, Yuanlong

    2012-11-01

    Using hydrographic measurements from three recent surveys in the western tropical Pacific, this study revealed the existence and general features of thermohaline finestructure near the northern Philippine coast. Pronounced finestructures were detected in the layers of the North Pacific Tropical Water (NPTW) and the North Pacific Intermediate Water (NPIW) during all three cruises and shown to be mainly thermohaline intrusions. Characteristics of the intrusions were further investigated with spiciness curvature and salinity anomaly methods. The vertical scale of the intrusions was 20-50 m and 50-100 m in the NPTW and NPIW layers, respectively. Within the NPTW layer, the Turner angle distribution and correlation between salinity and density anomalies suggested that diffusive convection between surface fresh water and subsurface saline water played an important role in the development and maintenance of the intrusions. In addition, connection between thermohaline finestructure and larger-scale oceanic processes was explored using historical hydrographic data. The results reveal that the salinity field and the distribution of the intrusions in this region were largely determined by mesoscale eddies. As a result of eddy stirring, both isopycnal and diapycnal temperature/salinity gradients were strengthened, which gave rise to the development of thermohaline intrusions. The intrusions acted to enhance heat and salt fluxes and resulted in the mixing of water masses being more efficient. By linking mesoscale eddy stirring to micro-scale diffusion, thermohaline finestructure plays a vital role in the ocean energy cascade and water mass conversion in the northern Philippine Sea.

  7. Northern Native Languages Project. Final Report.

    ERIC Educational Resources Information Center

    Burnaby, Barbara; And Others

    The Northern Native Language Project was constituted in June 1979 to produce a report which would present information on the present situation regarding language education schools in the project area of Ontario (James Bay, Nakina, and Sioux Lookout) and to make recommendations concerning appropriate action for the future. The introduction of the…

  8. A Grammar of Northern and Southern Gumuz

    ERIC Educational Resources Information Center

    Ahland, Colleen Anne

    2012-01-01

    Gumuz is a Nilo-Saharan dialect cluster spoken in the river valleys of northwestern Ethiopia and the southeastern part of the Republic of the Sudan. There are approximately 200,000 speakers, the majority of which reside in Ethiopia. This study is a phonological and grammatical analysis of two main dialects/languages: Northern Gumuz and Southern…

  9. From Poetry to Music: "Northern Lullaby"

    ERIC Educational Resources Information Center

    Cardany, Audrey Berger

    2011-01-01

    Nancy White Carlstrom's children's book, "Northern Lullaby," conjures through poetry the beauty of the Alaskan landscape in the evening. The book provides an opportunity for music teachers to help their students transform text and visual images to music. The author describes connections for reading comprehension in the general music classroom and…

  10. VHF radar measurements over Andoya (Northern Norway)

    NASA Technical Reports Server (NTRS)

    Czechowsky, P.; Reid, I. M.; Ruester, R.; Schmidt, G.

    1989-01-01

    The Mobile SOUSY Radar was operated during the MAP/WINE, the MAC/SINE, and MAC/Epsilon campaigns at Andoya in Northern Norway. A comparison between summer and winter results is presented, in particular the generation and development of the scattering regions, the different power spectral densities and the aspect sensitivities which were derived from six different beam directions.

  11. Winter in Northern Europe (WINE) Project

    NASA Technical Reports Server (NTRS)

    Vonzahn, U.

    1982-01-01

    The scientific aims, work plan, and organization of the Middle Atmosphere Program winter in northern Europe (MAP/WINE) are described. Proposed contributions to the MAP/WINE program from various countries are enumerated. Specific atmospheric parameters to be examined are listed along with the corresponding measurement technique.

  12. Regional health library service in northern Ireland.

    PubMed

    Crawford, D S

    1990-10-01

    The regional medical library service provided to physicians, hospitals, nurses, social workers, and health care administrators throughout Northern Ireland by the Queen's University of Belfast is described. A brief outline of the National Health Service in the United Kingdom is given, and the library service is described in terms of collections, cataloging, interlibrary loan, and reference. PMID:2224299

  13. Construction Services at Northern Arizona University.

    ERIC Educational Resources Information Center

    Van Dyke, Gary

    Construction Services is an innovative response to a chronic construction-remodeling problem at Northern Arizona State University. It is an in-house facilities maintenance department designed to address a variety of needs: prevention of construction or remodeling done by individual staff or faculty members without regard for applicable codes;…

  14. Nutritional condition of Northern Yellowstone Elk

    USGS Publications Warehouse

    Cook, R.C.; Cook, J.G.; Mech, L.D.

    2004-01-01

    Ultrasonography and body condition scoring was used to estimate nutritional condition of northern Yellowstone elk in late winter. Probability of pregnancy was related to body fat, and lactating cows had 50% less fat than non-lactating cows. For mild to normal winters, most of the elk were in good condition.

  15. Variation and Change in Northern Bavarian Quantity

    ERIC Educational Resources Information Center

    Drake, Derek

    2013-01-01

    This dissertation presents new research on the "Bavarian Quantity Law" (the BQL) in the northern Bavarian dialect of Hahnbach. Building upon earlier investigation of the BQL (cf. Bannert 1976a,b for Central Bavarian) this study examines the historical, phonological, and phonetic motivations for this feature as well the variability in its…

  16. 27 CFR 9.70 - Northern Sonoma.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... County-Lake County line. (24) The boundary follows the Sonoma County-Lake County line northwesterly to... the Northern Sonoma viticultural area are titled: (1) Sonoma County, California, scale 1:100 000, 1970..., California—Sonoma County; scale 1:24 000, 1955, photorevised 1975; (4) Camp Meeker Quadrangle,...

  17. 27 CFR 9.70 - Northern Sonoma.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... County-Lake County line. (24) The boundary follows the Sonoma County-Lake County line northwesterly to... the Northern Sonoma viticultural area are titled: (1) Sonoma County, California, scale 1:100 000, 1970..., California—Sonoma County; scale 1:24 000, 1955, photorevised 1975; (4) Camp Meeker Quadrangle,...

  18. 27 CFR 9.70 - Northern Sonoma.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... County-Lake County line. (24) The boundary follows the Sonoma County-Lake County line northwesterly to... the Northern Sonoma viticultural area are titled: (1) Sonoma County, California, scale 1:100 000, 1970..., California—Sonoma County; scale 1:24 000, 1955, photorevised 1975; (4) Camp Meeker Quadrangle,...

  19. Calibrating northern Texas High Plains groundwater model

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In the Northern High Plains of Texas, irrigated crop production accounts for a major portion of groundwater withdrawals from the Ogallala aquifer. The concern is that diminishing groundwater supplies will severely reduce regional crop and animal production, which in turn would impact the regional ec...

  20. Regional health library service in northern Ireland.

    PubMed Central

    Crawford, D S

    1990-01-01

    The regional medical library service provided to physicians, hospitals, nurses, social workers, and health care administrators throughout Northern Ireland by the Queen's University of Belfast is described. A brief outline of the National Health Service in the United Kingdom is given, and the library service is described in terms of collections, cataloging, interlibrary loan, and reference. PMID:2224299

  1. Governance and Aboriginal Claims in Northern Canada.

    ERIC Educational Resources Information Center

    Cozzetto, Don

    1990-01-01

    Focuses on problems of organization and governance that may follow settlement of Canadian aboriginal land claims. Compares financial problems, cultural issues such as subsistence lifestyles, and intergovernmental relations following the Alaska Native Claims Settlement Act, James Bay and Northern Quebec Agreement, and Western Arctic (Inuvialuit)…

  2. Applied Indigenous Studies at Northern Arizona University.

    ERIC Educational Resources Information Center

    Trosper, Ronald L.

    2001-01-01

    The Applied Indigenous Studies program at Northern Arizona University aims to prepare American Indian students to assume tribal leadership roles. Its location in the College of Ecosystem Science and Management emphasizes its land-oriented and applied focus. The program's development, core courses, and academic requirements for bachelors degrees…

  3. Orality in Northern Cree Indigenous Worlds.

    ERIC Educational Resources Information Center

    Weber-Pillwax, Cora

    2001-01-01

    Examines the importance and centrality of orality, rather than literacy, in the shared lives of the Cree of northern Alberta. Discusses orality consciousness related to the practice of shared memories and personal and communal healing during the "dance of the ancestors" or "ghost dance." Includes a short history of the Cree people and their…

  4. Diapause in northern corn rootworm (Coleoptera: Chrysomelidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Diabroticite corn rootworms are prominent pests of maize and have adapted to both cultural and chemical management methods. In response to a widely used corn-soybean crop rotation in the U.S. Corn Belt over several years, northern corn rootworm (NCR) populations adapted by increasing the proportion ...

  5. High Moisture Corn Evaluated for Northern Maine

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Profitable rotation crops that can grow in cool, northern Maine climates are needed to sustain the diversity of potato systems. A field experiment was conducted to determine whether three high moisture corn hybrids were suitable for harvest as a short season rotation crop. Yield and grain moisture...

  6. NORTHERN GUAM SOLE SOURCE AQUIFER- DESIGNATED AREA

    EPA Science Inventory

    Northern Guam Sole Source Aquifer Designated Area: In the Territory of Guam, entirely contained within the island of Guam. Specifically, that area within the coastal boundary defined by the line of Mean High water and north of the Adelup-Pago fault, plus some area south of the f...

  7. PERFORMANCE OF AERATED LAGOONS IN NORTHERN CLIMATES

    EPA Science Inventory

    Studies of cold climate aerated lagoons conducted by the Arctic Environmental Research Station, Fairbanks, Alaska are reported. Conclusions are based on these studies, observations of full scale aerated lagoons operating in Alaska and reports on lagoons in the northern tier of th...

  8. Ecohydrology Controls on Feedbacks Between Northern Wetlands and Climate Systems

    NASA Astrophysics Data System (ADS)

    Turetsky, M.; Harden, J.; McGuire, A.; Waddington, J.

    2007-12-01

    Boreal regions contain large stocks of soil carbon, mostly in poorly drained areas where peat accumulating wetlands have served as a long-term sink for atmospheric carbon. It is unknown whether northern wetlands globally will continue to represent a net sink for atmospheric carbon dioxide, or whether changes in the Earth's climate will cause these ecosystems to release stored carbon back to the atmosphere. Such feedbacks between northern wetlands and regional or global climate systems will depend on interactions between wetland vegetation, peat properties, and hydrology. Within many wetlands, hydrology is the dominant control on plant community structure and decomposition rates. In turn, both plant and microbial activity determine the quantity and quality of litter, which govern the nature of peat accumulation and soil properties critical to hydrology. Here, we will present research from our field and modeling studies investigating the effects of drought, permafrost degradation, and wildfire on vegetation, carbon cycling, and hydrological processes in northern wetlands at multiple spatial scales. At local scales, our findings show that interactions among vegetation, soil, and hydrology can lead to unexpected and often complex changes in soil environments, with potential 'carbon surprises'. For example, in a nonpermafrost peatland, we found that sustained drought led to peat subsidence that limited the development of oxic surface peat layers and inhibited ecosystem respiration. The decrease in porosity and water content with drought reduced seasonal ice thaw, which also likely limited microbial activity. In contrast, peatlands underlain by permafrost are increasingly experiencing thermokarst and soil flooding with increasing active layer depth. Changes in moss productivity post-thaw led to increased rates of organic matter accumulation, with very different hydrologic and soil properties than peat accumulated in permafrost settings. In addition to local

  9. Aerial view of reroofing of northern LTA ship hangar, circa ...

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

    Aerial view of re-roofing of northern LTA ship hangar, circa 1957. - Marine Corps Air Station Tustin, Northern Lighter Than Air Ship Hangar, Meffett Avenue & Maxfield Street, Tustin, Orange County, CA

  10. LOOKING NORTHEAST TOWARD NORTHERN AVE. SWING BRIDGE. BOSTON TEA PARTY ...

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

    LOOKING NORTHEAST TOWARD NORTHERN AVE. SWING BRIDGE. BOSTON TEA PARTY SHIP AT ANCHOR IN FOREGROUND. - Northern Avenue Swing Bridge, Spanning Fort Point Channel at boundary between Boston & South Boston, Boston, Suffolk County, MA

  11. 65. Photocopy of drawing from print, Burlington Northern Engineering Office) ...

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

    65. Photocopy of drawing from print, Burlington Northern Engineering Office) CENTER TOWER AND PORTALS - Burlington Northern Railroad Bridge, Spanning Willamette River at River Mile 6.9, Portland, Multnomah County, OR

  12. 47. Photocopy of photograph (from polaroid snapshot in Burlington Northern ...

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

    47. Photocopy of photograph (from polaroid snapshot in Burlington Northern Railroad correspondence files, October, 1957) SHEAR FENCE DAMAGE FROM M/V KORSHOLMA - Burlington Northern Railroad Bridge, Spanning Willamette River at River Mile 6.9, Portland, Multnomah County, OR

  13. 49. Photocopy of photograph (from polaroid snapshot in Burlington Northern ...

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

    49. Photocopy of photograph (from polaroid snapshot in Burlington Northern Railroad correspondence files, 1957) PIER III DAMAGE FROM M/V KORSHOLMA COLLISION - Burlington Northern Railroad Bridge, Spanning Willamette River at River Mile 6.9, Portland, Multnomah County, OR

  14. 66. Photocopy of drawing (from print, Burlington Northern Engineering Office) ...

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

    66. Photocopy of drawing (from print, Burlington Northern Engineering Office) TURNTABLE BEARINGS AND LOADING BEAMS - Burlington Northern Railroad Bridge, Spanning Willamette River at River Mile 6.9, Portland, Multnomah County, OR

  15. 68. Photocopy of drawing (from print, Burlington Northern Engineering Office) ...

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

    68. Photocopy of drawing (from print, Burlington Northern Engineering Office) STRESS DIAGRAM - Burlington Northern Railroad Bridge, Spanning Willamette River at River Mile 6.9, Portland, Multnomah County, OR

  16. 64. Photocopy of drawing (from print, Burlington Northern Engineering Office, ...

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

    64. Photocopy of drawing (from print, Burlington Northern Engineering Office, Seattle) STRESS SHEET - Burlington Northern Railroad Bridge, Spanning Willamette River at River Mile 6.9, Portland, Multnomah County, OR

  17. Additional evidence of Mercurian volcanism

    USGS Publications Warehouse

    Trask, N.J.; Strom, R.G.

    1976-01-01

    Evidence concerned with (1) the character and distribution of terrain surrounding fresh basins, (2) albedo, color and temporal differences between a basin rim and smooth plains on its floor, and (3) the stratigraphic relations and local distribution of smooth plains in the hilly and lineated terrain are cited as additional evidence for an internal origin of much of the Mercurian smooth plains. Altough the question of Mercurian volcanism should be kept open, this evidence together with that presented in an earlier paper suggests that volcanism occurred on Mercury early in its history. ?? 1976.

  18. Individualized additional instruction for calculus

    NASA Astrophysics Data System (ADS)

    Takata, Ken

    2010-10-01

    College students enrolling in the calculus sequence have a wide variance in their preparation and abilities, yet they are usually taught from the same lecture. We describe another pedagogical model of Individualized Additional Instruction (IAI) that assesses each student frequently and prescribes further instruction and homework based on the student's performance. Our study compares two calculus classes, one taught with mandatory remedial IAI and the other without. The class with mandatory remedial IAI did significantly better on comprehensive multiple-choice exams, participated more frequently in classroom discussion and showed greater interest in theorem-proving and other advanced topics.

  19. Water based drilling mud additive

    SciTech Connect

    McCrary, J.L.

    1983-12-13

    A water based fluid additive useful in drilling mud used during drilling of an oil or gas well is disclosed, produced by reacting water at temperatures between 210/sup 0/-280/sup 0/ F. with a mixture comprising in percent by weight: gilsonite 25-30%, tannin 7-15%, lignite 25-35%, sulfonating compound 15-25%, water soluble base compound 5-15%, methylene-yielding compound 1-5%, and then removing substantially all of the remaining water to produce a dried product.

  20. Metal Additive Manufacturing: A Review

    NASA Astrophysics Data System (ADS)

    Frazier, William E.

    2014-06-01

    This paper reviews the state-of-the-art of an important, rapidly emerging, manufacturing technology that is alternatively called additive manufacturing (AM), direct digital manufacturing, free form fabrication, or 3D printing, etc. A broad contextual overview of metallic AM is provided. AM has the potential to revolutionize the global parts manufacturing and logistics landscape. It enables distributed manufacturing and the productions of parts-on-demand while offering the potential to reduce cost, energy consumption, and carbon footprint. This paper explores the material science, processes, and business consideration associated with achieving these performance gains. It is concluded that a paradigm shift is required in order to fully exploit AM potential.

  1. Caribbean basin framework, 2: Northern Central America

    SciTech Connect

    Tyburski, S.A.; Gordon, M.B.; Mann, P. )

    1991-03-01

    There are four Jurassic to Recent basin-forming periods in northern Central America (honduras, Honduran Borderlands, Belize, Guatemala, northern Nicaragua): (1) Middle Jurassic-Early Cretaceous rifting and subsidence along normal faults in Honduras and Guatemala; rifts are suggested but are not well defined in Honduras by the distribution of clastic sediments and associated volcanic rocks. Rifting is attributed to the separation of Central America from the southern margin of the North American plate; (2) Cretaceous subsidence recorded by the development of a Cretaceous carbonate platform in Honduras, Guatemala, and Belize; subsidence is attributed to thermal subsidence of the rifted margins of the various blocks; (3) Late Cretaceous-Recent development of a volcanic arc along the western margin of Middle America and the northern margin of Honduras; (4) Late Cretaceous large-scale folding in Honduras, ophiolite obduction, and formation of a foredeep basin in Guatemala (Sepur trough); deformation is attributed to the collision between a north-facing arc in northern Honduras and the Nicaraguan Rise and the passive margin of Guatemala and Belize; and (5) Eocene to Recent strike-slip faulting along the present-day North American-Caribbean plate boundary in Guatemala, northern Honduras, and Belize. Strike-slip faults and basins form a California-type borderlands characterized by elongate basins that appear as half-grabens in profile. Counterclockwise rotation of the central honduras plateau, a thicker and topographically higher-than-average block within the plate boundary zone, is accommodated by rifting or strike-slip faults at its edges.

  2. Matrix-assisted laser desorption-ionization mass spectrometry peptide mass fingerprinting for proteome analysis: identification efficiency after on-blot or in-gel digestion with and without desalting procedures.

    PubMed

    Lamer, S; Jungblut, P R

    2001-03-10

    In theory, peptide mass fingerprinting by matrix assisted laser desorption-ionization mass spectrometry (MALDI-MS) has the potential to identify all of the proteins detected by silver staining on gels. In practice, if the genome of the organism investigated is completely sequenced, using current techniques, all proteins stained by Coomassie Brilliant Blue can be identified. This loss of identification sensitivity of ten to hundred-fold is caused by loss of peptides by surface contacts. Therefore, we performed digestion and transfer of peptides in the lower microl range and reduced the number of steps. The peptide mix obtained from in-gel or on-blot digestion was analyzed directly after digestion or after concentration on POROS R2 beads. Eight protein spots of a 2-DE gel from Mycobacterium bovis BCG were identified using these four preparation procedures for MALDI-MS. Overall, on-blot digestion was as effective as in-gel digestion. Whereas higher signal intensities resulted after concentration, hydrophilic peptides are better detected by direct measurement of the peptide mix without POROS R2 concentration. PMID:11270870

  3. Expansion and methylation status at FRAXE can be detected on EcoRI blots used for FRAXA diagnosis: analysis of four FRAXE families with mild mental retardation in males.

    PubMed Central

    Biancalana, V.; Taine, L.; Bouix, J. C.; Finck, S.; Chauvin, A.; De Verneuil, H.; Knight, S. J.; Stoll, C.; Lacombe, D.; Mandel, J. L.

    1996-01-01

    The original test for the analysis of the CCG expansion at the FRAXE locus involves Southern blot analysis of HindIII digests. We show that, by using a different probe, the FRAXE mutation can be detected easily on the same EcoRI or EagI+EcoRI blots as are used for detection of FRAXA. Unexpectedly, we found that both the expansion and methylation status can be determined on a single EcoRI digest, because of the presence of a methylation-sensitive EcoRI site very close to the CCG repeat. We thus detected in a series of mentally retarded individuals previously tested for FRAXA expansion a FRAXE proband who led to the identification of a large sibship (7 of 10 children carrying a mutation). We also show that two fragile X families without FRAXA mutation that previously have been described by Oberlé et al. have the FRAXE expansion. In another family also ascertained initially by cytogenetic finding of a fragile X site, we performed the combined cytogenetic and molecular prenatal diagnosis of a mutated male fetus. All nine males (>3 years old) in whom we found a methylated mutation had mild mental retardation. Our results suggest that the threshold of repeat length for abnormal methylation and fragile-site expression may be smaller at FRAXE than at FRAXA. Images Figure 2 Figure 3 Figure 4 PMID:8808600

  4. Additive manufacturing of RF absorbers

    NASA Astrophysics Data System (ADS)

    Mills, Matthew S.

    The ability of additive manufacturing techniques to fabricate integrated electromagnetic absorbers tuned for specific radio frequency bands within structural composites allows for unique combinations of mechanical and electromagnetic properties. These composites and films can be used for RF shielding of sensitive electromagnetic components through in-plane and out-of-plane RF absorption. Structural composites are a common building block of many commercial platforms. These platforms may be placed in situations in which there is a need for embedded RF absorbing properties along with structural properties. Instead of adding radar absorbing treatments to the external surface of existing structures, which adds increased size, weight and cost; it could prove to be advantageous to integrate the microwave absorbing properties directly into the composite during the fabrication process. In this thesis, a method based on additive manufacturing techniques of composites structures with prescribed electromagnetic loss, within the frequency range 1 to 26GHz, is presented. This method utilizes screen printing and nScrypt micro dispensing to pattern a carbon based ink onto low loss substrates. The materials chosen for this study will be presented, and the fabrication technique that these materials went through to create RF absorbing structures will be described. The calibration methods used, the modeling of the RF structures, and the applications in which this technology can be utilized will also be presented.

  5. High Flow Addition Curing Polyimides

    NASA Technical Reports Server (NTRS)

    Chuang, Kathy C.; Vannucci, Raymond D.; Ansari, Irfan; Cerny, Lawrence L.; Scheiman, Daniel A.

    1994-01-01

    A new series of high flow PMR-type addition curing polyimides was developed, which employed the substitution of 2,2'-bis (trifluoromethyl) -4,4'-diaminobiphenyl (BTDB) for p-phenylenediamine (p -PDA) in a PMR-IL formulation. These thermoset polyimides, designated as 12F resins, were prepared from BTDB and the dimethyl ester of 4,4'- (hexafluo- roisopropylidene) -diphthalic acid (HFDE) with either nadic ester (NE) or p-aminostyrene (PAS) as the endcaps for addition curing. The 12F prepolymers displayed lower melting temperatures in DSC analysis, and higher melt flow in rheological studies than the cor- responding PMR-11 polyimides. Long-term isothermal aging studies showed that BTDB- based 12F resins exhibited comparable thermo-oxidative stability to P-PDA based PMR-11 polyimides. The noncoplanar 2- and 2'-disubstituted biphenyldiamine (BTDB) not only lowered the melt viscosities of 12F prepolymers, but also retained reasonable thermal sta- bility of the cured resins. The 12F polyimide resin with p-aminostyrene endcaps showed the best promise for long-term, high-temperature application at 343 C (650 F).

  6. Improvement of Lyme borreliosis serodiagnosis by a newly developed recombinant immunoglobulin G (IgG) and IgM line immunoblot assay and addition of VlsE and DbpA homologues.

    PubMed

    Goettner, Gereon; Schulte-Spechtel, Ulrike; Hillermann, Ruth; Liegl, Gabi; Wilske, Bettina; Fingerle, Volker

    2005-08-01

    We developed and evaluated a recombinant Borrelia line immunoblot assay based on 18 homologues of seven different antigens, i.e., p100, p58, p41i, BmpA, VlsE, OspC, and DbpA. Each recombinant antigen can be detected separately and is distinct even from homologues with identical molecular weights. This blot was compared to the recently described recombinant Borrelia Western immunoblot assay (U. Schulte-Spechtel, G. Lehnert, G. Liegl, V. Fingerle, C. Heimerl, B. J. Johnson, and B. Wilske, J. Clin. Microbiol. 41:1299-1303, 2003). To verify sensitivity and specificity, both blots were evaluated for reactivity with Borrelia-specific immunoglobulin G (IgG) and IgM antibodies with 85 sera from patients with different manifestations of Lyme borreliosis and 110 controls. According to European interpretation criteria for Borrelia Western blots, which define a serum as positive when it recognizes at least two bands, sensitivity increased significantly from 70.6% (Western blot) to 84.7% (line blot) for IgG (P = 0.042) and from 40.0% (Western blot) to 73.8% (line blot) for IgM (P < 0.005). The increased sensitivity for IgG detection is due to the new line blot technique, whereas the improvement in detection of IgM is mainly achieved through incorporation of the additional antigens. Notably, the recombinant VlsE of Borrelia garinii strain PBi displayed the highest sensitivity of all antigens tested for IgG detection and is also one of the most useful antigens for IgM. Due to its excellent sensitivity and specificity combined with ease of evaluation, this line immunoblot assay offers a useful improvement in serodiagnosis of Lyme borreliosis. PMID:16081885

  7. Annotated bibliography of the Black Warrior basin area, northern Alabama - northern Mississippi

    SciTech Connect

    Ward-McLemore, E.

    1983-01-01

    This bibliography contains 1964 records related to the geology of the Black Warrior basin of northern Alabama and northern Mississippi. Specific topics include, but are not limited to: coal, petroleum, and natural gas deposits; mineralogy; lithology; paleontology; petrology; stratigraphy; tectonics; bauxite; iron ores; geologic correlations; earthquakes; fossils; gold deposits; geological surveys; hydrology; and water resources. The subject index provides listings of records related to each county and the geologic ages covered by this area. Some of the items (54) are themselves bibliographies.

  8. AmeriFlux CA-Man Manitoba - Northern Old Black Spruce (former BOREAS Northern Study Area)

    SciTech Connect

    Amiro, Brian

    2016-01-01

    This is the AmeriFlux version of the carbon flux data for the site CA-Man Manitoba - Northern Old Black Spruce (former BOREAS Northern Study Area). Site Description - 55.880° N, 98.481° W, elevation of 259 m, Boreal coniferous: Black spruce; occasional larch present in poorly-drained areas. Groundcover is moss (feathermosses and Sphagnum), Labrador Tea, Vaccinium, and willows are a main component of the understory. It was established in 1993 as a BOREAS site.

  9. Osteogenesis Imperfecta Type VI in Individuals from Northern Canada.

    PubMed

    Ward, Leanne; Bardai, Ghalib; Moffatt, Pierre; Al-Jallad, Hadil; Trejo, Pamela; Glorieux, Francis H; Rauch, Frank

    2016-06-01

    Osteogenesis imperfecta (OI) type VI is a recessively inherited form of OI that is caused by mutations in SERPINF1, the gene coding for pigment-epithelium derived factor (PEDF). Here, we report on two apparently unrelated children with OI type VI who had the same unusual homozygous variant in intron 6 of SERPINF1 (c.787-10C>G). This variant created a novel splice site that led to the in-frame addition of three amino acids to PEDF (p.Lys262_Ile263insLeuSerGln). Western blotting showed that skin fibroblasts with this mutation produced PEDF but failed to secrete it. Both children were treated with intravenous bisphosphonates, but the treatment of Individual 1 was switched to subcutaneous injections of denosumab (dose 1 mg per kg body weight, repeated every 3 months). An iliac bone sample obtained after 5 denosumab injections (and 3 months after the last injection) showed no change in the increased osteoid parameters that are typical of OI type VI, but the number of osteoclasts in trabecular bone was markedly increased. This suggests that the effect of denosumab on osteoclast suppression is of shorter duration in children with OI type VI than what has previously been reported on adults with osteoporosis. PMID:26815784

  10. 76 FR 44625 - Northern Lights Variable Trust, et al.;

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-07-26

    ... COMMISSION Northern Lights Variable Trust, et al.; Notice of Application July 19, 2011. AGENCY: Securities.... APPLICANTS: Northern Lights Variable Trust (the ``Fund'') and Gemini Fund Services, LLC (``Gemini.... Northern Lights Variable Trust, c/o Emile Molineaux, Esquire, Gemini Fund Services, LLC, 450...

  11. 27 CFR 9.109 - Northern Neck George Washington Birthplace.

    Code of Federal Regulations, 2012 CFR

    2012-04-01

    ... Viticultural Areas § 9.109 Northern Neck George Washington Birthplace. (a) Name. The name of the viticultural area described in this section is “Northern Neck George Washington Birthplace.” (b) Approved maps. The appropriate maps for determining the boundaries of the Northern Neck George Washington Birthplace...

  12. 27 CFR 9.109 - Northern Neck George Washington Birthplace.

    Code of Federal Regulations, 2014 CFR

    2014-04-01

    ... Viticultural Areas § 9.109 Northern Neck George Washington Birthplace. (a) Name. The name of the viticultural area described in this section is “Northern Neck George Washington Birthplace.” (b) Approved maps. The appropriate maps for determining the boundaries of the Northern Neck George Washington Birthplace...

  13. 27 CFR 9.109 - Northern Neck George Washington Birthplace.

    Code of Federal Regulations, 2011 CFR

    2011-04-01

    ... Viticultural Areas § 9.109 Northern Neck George Washington Birthplace. (a) Name. The name of the viticultural area described in this section is “Northern Neck George Washington Birthplace.” (b) Approved maps. The appropriate maps for determining the boundaries of the Northern Neck George Washington Birthplace...

  14. 27 CFR 9.109 - Northern Neck George Washington Birthplace.

    Code of Federal Regulations, 2010 CFR

    2010-04-01

    ... Viticultural Areas § 9.109 Northern Neck George Washington Birthplace. (a) Name. The name of the viticultural area described in this section is “Northern Neck George Washington Birthplace.” (b) Approved maps. The appropriate maps for determining the boundaries of the Northern Neck George Washington Birthplace...

  15. 27 CFR 9.109 - Northern Neck George Washington Birthplace.

    Code of Federal Regulations, 2013 CFR

    2013-04-01

    ... Viticultural Areas § 9.109 Northern Neck George Washington Birthplace. (a) Name. The name of the viticultural area described in this section is “Northern Neck George Washington Birthplace.” (b) Approved maps. The appropriate maps for determining the boundaries of the Northern Neck George Washington Birthplace...

  16. 8 CFR 235.9 - Northern Marianas identification card.

    Code of Federal Regulations, 2011 CFR

    2011-01-01

    ... 8 Aliens and Nationality 1 2011-01-01 2011-01-01 false Northern Marianas identification card. 235... INSPECTION OF PERSONS APPLYING FOR ADMISSION § 235.9 Northern Marianas identification card. During the two-year period that ended July 1, 1990, the Service issued Northern Marianas Identification Cards...

  17. 8 CFR 235.9 - Northern Marianas identification card.

    Code of Federal Regulations, 2012 CFR

    2012-01-01

    ... 8 Aliens and Nationality 1 2012-01-01 2012-01-01 false Northern Marianas identification card. 235... INSPECTION OF PERSONS APPLYING FOR ADMISSION § 235.9 Northern Marianas identification card. During the two-year period that ended July 1, 1990, the Service issued Northern Marianas Identification Cards...

  18. 8 CFR 235.9 - Northern Marianas identification card.

    Code of Federal Regulations, 2013 CFR

    2013-01-01

    ... 8 Aliens and Nationality 1 2013-01-01 2013-01-01 false Northern Marianas identification card. 235... INSPECTION OF PERSONS APPLYING FOR ADMISSION § 235.9 Northern Marianas identification card. During the two-year period that ended July 1, 1990, the Service issued Northern Marianas Identification Cards...

  19. 8 CFR 235.9 - Northern Marianas identification card.

    Code of Federal Regulations, 2014 CFR

    2014-01-01

    ... 8 Aliens and Nationality 1 2014-01-01 2014-01-01 false Northern Marianas identification card. 235... INSPECTION OF PERSONS APPLYING FOR ADMISSION § 235.9 Northern Marianas identification card. During the two-year period that ended July 1, 1990, the Service issued Northern Marianas Identification Cards...

  20. 8 CFR 235.9 - Northern Marianas identification card.

    Code of Federal Regulations, 2010 CFR

    2010-01-01

    ... 8 Aliens and Nationality 1 2010-01-01 2010-01-01 false Northern Marianas identification card. 235... INSPECTION OF PERSONS APPLYING FOR ADMISSION § 235.9 Northern Marianas identification card. During the two-year period that ended July 1, 1990, the Service issued Northern Marianas Identification Cards...

  1. 78 FR 51716 - Northern Natural Gas Company; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-08-21

    ... Energy Regulatory Commission Northern Natural Gas Company; Notice of Application Take notice that on August 1, 2013, Northern Natural Gas Company (Northern), 1111 South 103rd Street, Omaha, Nebraska 68124, filed an application pursuant to section 7(c) of the Natural Gas Act and part 157 of the...

  2. 78 FR 8501 - Northern Natural Gas Company; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2013-02-06

    ... Energy Regulatory Commission Northern Natural Gas Company; Notice of Application Take notice that on January 18, 2013, Northern Natural Gas Company (Northern), 1111 South 103rd Street, Omaha, Nebraska 68124, filed in Docket No. CP13-53-000, an application pursuant to section 7(c) of the Natural Gas Act...

  3. 77 FR 35958 - Northern Natural Gas Company; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-06-15

    ... Energy Regulatory Commission Northern Natural Gas Company; Notice of Application Take notice that on May 30, 2012, Northern Natural Gas Company (Northern), 1111 South 103rd Street, Omaha, Nebraska 68124... regulations and section 7(b) of the Natural Gas Act, to abandon by sale to DKM Enterprises, LLC (DKM)...

  4. 76 FR 12721 - Northern Natural Gas Company; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2011-03-08

    ... Energy Regulatory Commission Northern Natural Gas Company; Notice of Application Take notice that on February 18, 2011, Northern Natural Gas Company (Northern), 1111 South 103 Street, Omaha, Nebraska 68124-1000, filed in Docket No. CP11-98-000, an application pursuant to section 7(b) of the Natural Gas...

  5. 75 FR 35779 - Northern Natural Gas Company; Notice of Application

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-06-23

    ... Energy Regulatory Commission Northern Natural Gas Company; Notice of Application June 16, 2010. Take notice that on June 2, 2010, Northern Natural Gas Company (Northern), 1111 South 103rd Street, Omaha... Natural Gas Act, for a certificate of public convenience and necessity authorizing the increase...

  6. Importance of Nitrogen Availability on Land Carbon Sequestration in Northern Eurasia during the 21st Century

    NASA Astrophysics Data System (ADS)

    Kicklighter, D. W.; Melillo, J. M.; Monier, E.; Sokolov, A. P.; Lu, X.; Zhuang, Q.

    2015-12-01

    Atmospheric nitrogen deposition, nitrogen fixation, and the application of nitrogen fertilizers provide subsidies to land ecosystems that can increase nitrogen availability for vegetation production and thereby influence land carbon dynamics. In addition, enhanced decomposition of soil organic matter (SOM) from warming soils and permafrost degradation may also increase nitrogen availability in Northern Eurasia. Here, we examine how changes in nitrogen availability may influence land carbon dynamics in Northern Eurasia during the 21st century by comparing results for a "business as usual" scenario (the IPCC Representative Concentration Pathways or RCP 8.5) and a stabilization scenario (RCP 4.5) between a version of the Terrestrial Ecosystem Model that does not consider the effects of atmospheric nitrogen deposition, nitrogen fixation and soil thermal dynamics on land carbon dynamics (TEM 4.4) and a version that does consider these dynamics (TEM 6.0). In these simulations, atmospheric nitrogen deposition, nitrogen fixation, and fertilizer applications provide an additional 3.3 Pg N (RCP 4.5) to 3.9 Pg N (RCP 8.5) to Northern Eurasian ecosystems over the 21st century. Land ecosystems retain about 38% (RCP4.5) to 48% (RCP 8.5) of this nitrogen subsidy. Net nitrogen mineralization estimated by TEM 6.0 provide an additional 1.0 Pg N to vegetation than estimated by TEM 4.4 over the 21st century from enhanced decomposition of SOM including SOM formerly protected by permafrost. The enhanced nitrogen availability in TEM 6.0 allows Northern Eurasian ecosystems to sequester 1.8x (RCP 8.5) to 2.4x (RCP 4.5) more carbon over the 21st century than estimated by TEM 4.4. Our results indicate that consideration of nitrogen subsidies and soil thermal dynamics have a large influence on how simulated land carbon dynamics in Northern Eurasia will respond to future changes in climate, atmospheric chemistry, and disturbances.

  7. Neutron Characterization for Additive Manufacturing

    NASA Technical Reports Server (NTRS)

    Watkins, Thomas; Bilheux, Hassina; An, Ke; Payzant, Andrew; DeHoff, Ryan; Duty, Chad; Peter, William; Blue, Craig; Brice, Craig A.

    2013-01-01

    Oak Ridge National Laboratory (ORNL) is leveraging decades of experience in neutron characterization of advanced materials together with resources such as the Spallation Neutron Source (SNS) and the High Flux Isotope Reactor (HFIR) shown in Fig. 1 to solve challenging problems in additive manufacturing (AM). Additive manufacturing, or three-dimensional (3-D) printing, is a rapidly maturing technology wherein components are built by selectively adding feedstock material at locations specified by a computer model. The majority of these technologies use thermally driven phase change mechanisms to convert the feedstock into functioning material. As the molten material cools and solidifies, the component is subjected to significant thermal gradients, generating significant internal stresses throughout the part (Fig. 2). As layers are added, inherent residual stresses cause warping and distortions that lead to geometrical differences between the final part and the original computer generated design. This effect also limits geometries that can be fabricated using AM, such as thin-walled, high-aspect- ratio, and overhanging structures. Distortion may be minimized by intelligent toolpath planning or strategic placement of support structures, but these approaches are not well understood and often "Edisonian" in nature. Residual stresses can also impact component performance during operation. For example, in a thermally cycled environment such as a high-pressure turbine engine, residual stresses can cause components to distort unpredictably. Different thermal treatments on as-fabricated AM components have been used to minimize residual stress, but components still retain a nonhomogeneous stress state and/or demonstrate a relaxation-derived geometric distortion. Industry, federal laboratory, and university collaboration is needed to address these challenges and enable the U.S. to compete in the global market. Work is currently being conducted on AM technologies at the ORNL

  8. Additives in fibers and fabrics.

    PubMed

    Barker, R H

    1975-06-01

    The additives and contaminants which occur in textile fibers vary widely, depending on the type of fiber and the pretreatment which it has received. Synthetic fibers such as nylon and polyester contain trace amounts of contaminants such as catalysts and catalyst deactivators which remain after the synthesis of the basic polymers. In addition, there are frequently a number of materials which are added to perform specific functions in almost all man-made fibers. Examples of these would include traces of metals or metal salts used as tracers for identification of specific lots of fiber, TiO2 or similar materials added as delustrants, and a host of organic species added for such special purposes as antistatic agents or flame retardants. There may also be considerable quantities of residual monomer or small oligomers dissolved in the polymer matrix. The situation becomes even more complex after the fibers are converted into fabric form. Numerous materials are applied at various stages of fabric preparation to act as lubricants, sizing agents, antistats, bleaches, and wetting agents to facilitate the processing, but these are normally removed before the fabric reaches the cutters of the ultimate consumers and therefore usually do not constitute potential hazards. However, there are many other chemical agents which are frequently added during the later stages of fabric preparation and which are not designed to be removed. Aside from dyes and printing pigments, the most common additive for apparel fabrics is a durable press treatment. This generally involves the use of materials capable of crosslinking cellulosics by reacting through such functions as N-methylolated amides or related compounds such as ureas and carbamates. These materials pose some potential hazards due to both the nitrogenous bases and the formaldehyde which they usually release. There is usually also some residual catalyst in fabrics which have received such treatments. Other types of chemical treatments

  9. Additives in fibers and fabrics.

    PubMed Central

    Barker, R H

    1975-01-01

    The additives and contaminants which occur in textile fibers vary widely, depending on the type of fiber and the pretreatment which it has received. Synthetic fibers such as nylon and polyester contain trace amounts of contaminants such as catalysts and catalyst deactivators which remain after the synthesis of the basic polymers. In addition, there are frequently a number of materials which are added to perform specific functions in almost all man-made fibers. Examples of these would include traces of metals or metal salts used as tracers for identification of specific lots of fiber, TiO2 or similar materials added as delustrants, and a host of organic species added for such special purposes as antistatic agents or flame retardants. There may also be considerable quantities of residual monomer or small oligomers dissolved in the polymer matrix. The situation becomes even more complex after the fibers are converted into fabric form. Numerous materials are applied at various stages of fabric preparation to act as lubricants, sizing agents, antistats, bleaches, and wetting agents to facilitate the processing, but these are normally removed before the fabric reaches the cutters of the ultimate consumers and therefore usually do not constitute potential hazards. However, there are many other chemical agents which are frequently added during the later stages of fabric preparation and which are not designed to be removed. Aside from dyes and printing pigments, the most common additive for apparel fabrics is a durable press treatment. This generally involves the use of materials capable of crosslinking cellulosics by reacting through such functions as N-methylolated amides or related compounds such as ureas and carbamates. These materials pose some potential hazards due to both the nitrogenous bases and the formaldehyde which they usually release. There is usually also some residual catalyst in fabrics which have received such treatments. Other types of chemical treatments

  10. 75 FR 62807 - Northern Border Pipeline Company; Notice of Availability of the Environmental Assessment for the...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2010-10-13

    ... Energy Regulatory Commission Northern Border Pipeline Company; Notice of Availability of the... Princeton Lateral Project proposed by Northern Border Pipeline Company (Northern Border) in the above referenced docket. Northern Border requests authorization to construct pipeline facilities to...

  11. A novel addition polyimide adhesive

    NASA Technical Reports Server (NTRS)

    St.clair, T. L.; Progar, D. J.

    1981-01-01

    An addition polyimide adhesive, LARC 13, was developed which shows promise for bonding both titanium and composites for applications which require service temperatures in excess of 533 K. The LARC 13 is based on an oligomeric bis nadimide containing a meta linked aromatic diamine. The adhesive melts prior to polymerization due to its oligomeric nature, thereby allowing it to be processed at 344 kPa or less. Therefore, LARC 13 is ideal for the bonding of honeycomb sandwich structures. After melting, the resin thermosets during the cure of the nadic endcaps to a highly crosslinked system. Few volatiles are evolved, thus allowing large enclosed structures to be bonded. Preparation of the adhesive as well as bonding, aging, and testing of lap shear and honeycomb samples are discussed.

  12. Optics of progressive addition lenses.

    PubMed

    Sheedy, J E; Buri, M; Bailey, I L; Azus, J; Borish, I M

    1987-02-01

    The optical characteristics of the major progressive addition lenses were measured using an automated lensometer with a specially designed lens holder to simulate eye rotation. Measurements were made every 3 degrees (about 1.5 mm) and graphs of isospherical equivalent lines and isocylinder lines were developed. Generally the near zone of these lenses is narrower and lower than in bifocal or trifocal lenses. Distinct differences exist between the various progressive lenses. The width of the near zone, rate of power progression, amount of unwanted cylinder (level with the distance center), and clarity of the distance zone are compared for the various lenses. The optical measurements demonstrate an apparent trade-off between the size of the cylinder-free area of the lens and the amount of the cylinder. PMID:3826294

  13. Addition polyimide end cap study

    NASA Technical Reports Server (NTRS)

    St.clair, T. L.

    1980-01-01

    The characterization of addition polyimides with various end caps for adhesive applications at 120-250 C environments is discussed. Oligometric polyimides were prepared from 3,3',4,4'-benzophenone tetracarboxylic dianhydride and 3,3'-methylenedianiline which were end-capped with functionally reactive moities which cause crosslinking when the oligomers are heated to 200-400 C. The syntheses of the oligomers are outlined. The thermolysis of the oligomers was studied by differential scanning calorimetry and the resulting polymers were characterized by differential thermal analysis and adhesive performance. The adhesive data include lap shear strengths on titanium 6-4 adherends both before and after aging for 1000 hours at 121 C and/or 232 C.

  14. An Additive Manufacturing Test Artifact

    PubMed Central

    Moylan, Shawn; Slotwinski, John; Cooke, April; Jurrens, Kevin; Donmez, M Alkan

    2014-01-01

    A test artifact, intended for standardization, is proposed for the purpose of evaluating the performance of additive manufacturing (AM) systems. A thorough analysis of previously proposed AM test artifacts as well as experience with machining test artifacts have inspired the design of the proposed test artifact. This new artifact is designed to provide a characterization of the capabilities and limitations of an AM system, as well as to allow system improvement by linking specific errors measured in the test artifact to specific sources in the AM system. The proposed test artifact has been built in multiple materials using multiple AM technologies. The results of several of the builds are discussed, demonstrating how the measurement results can be used to characterize and improve a specific AM system. PMID:26601039

  15. An Additive Manufacturing Test Artifact.

    PubMed

    Moylan, Shawn; Slotwinski, John; Cooke, April; Jurrens, Kevin; Donmez, M Alkan

    2014-01-01

    A test artifact, intended for standardization, is proposed for the purpose of evaluating the performance of additive manufacturing (AM) systems. A thorough analysis of previously proposed AM test artifacts as well as experience with machining test artifacts have inspired the design of the proposed test artifact. This new artifact is designed to provide a characterization of the capabilities and limitations of an AM system, as well as to allow system improvement by linking specific errors measured in the test artifact to specific sources in the AM system. The proposed test artifact has been built in multiple materials using multiple AM technologies. The results of several of the builds are discussed, demonstrating how the measurement results can be used to characterize and improve a specific AM system. PMID:26601039

  16. SIPSEY WILDERNESS AND ADDITIONS, ALABAMA.

    USGS Publications Warehouse

    Schweinfurth, Stanley P.; Mory, Peter C.

    1984-01-01

    On the basis of geologic, geochemical, and mineral surveys the Sipsey Wilderness and additions are deemed to have little promise for the occurrence of metallic mineral resources. Although limestone, shale, and sandstone resources that occur in the area are physically suitable for a variety of uses, similar materials are available outside the area closer to transportation routes and potential markets. A small amount of coal has been identified in the area, occurring as nonpersistent beds less than 28 in. thick. Oil and (or) natural gas resources may be present if suitable structural traps exist in the subsurface. Therefore, the area has a probable oil and gas potential. Small amounts of asphaltic sandstone and limestone, commonly referred to as tar sands, may also occur in the subsurface. 5 refs.

  17. Adverse reactions to food additives.

    PubMed

    Simon, R A

    1986-01-01

    There are thousands of agents that are intentionally added to the food that we consume. These include preservatives, stabilizers, conditioners, thickeners, colorings, flavorings, sweeteners, antioxidants, etc. etc. Yet only a surprisingly small number have been associated with hypersensitivity reactions. Amongst all the additives, FD&C dyes have been most frequently associated with adverse reactions. Tartrazine is the most notorious of them all; however, critical review of the medical literature and current Scripps Clinic studies would indicate that tartrazine has been confirmed to be at best only occasionally associated with flares of urticaria or asthma. There is no convincing evidence in the literature of reactivity to the other azo or nonazo dyes. This can also be said of BHA/BHT, nitrites/nitrates and sorbates. Parabens have been shown to elicit IgE mediated hypersensitivity reactions when used as pharmaceutical preservatives; however, as with the other additives noted above, ingested parabens have only occasionally been associated with adverse reactions. MSG, the cause of the 'Chinese restaurant syndrome' has only been linked to asthma in one report. Sulfiting agents used primarily as food fresheners and to control microbial growth in fermented beverages have been established as the cause of any where from mild to severe and even fatal reactions in at least 5% of the asthmatic population. Other reactions reported to follow sulfite ingestion include anaphylaxis, gastro intestinal complaints and dermatological eruptions. The prevalence of these non asthmatic reactions is unknown. The mechanism of sulfite sensitive asthma is also unknown but most likely involves hyperreactivity to inhale SO2 in the great majority of cases; however, there are reports of IgE mediated reactions and other sulfite sensitive asthmatics have been found with low levels of sulfite oxidase; necessary to oxidize endogenous sulfite to sulfate. PMID:3302664

  18. Hubble Spots Northern Hemispheric Clouds on Uranus

    NASA Technical Reports Server (NTRS)

    1997-01-01

    Using visible light, astronomers for the first time this century have detected clouds in the northern hemisphere of Uranus. The newest images, taken July 31 and Aug. 1, 1997 with NASA Hubble Space Telescope's Wide Field and Planetary Camera 2, show banded structure and multiple clouds. Using these images, Dr. Heidi Hammel (Massachusetts Institute of Technology) and colleagues Wes Lockwood (Lowell Observatory) and Kathy Rages (NASA Ames Research Center) plan to measure the wind speeds in the northern hemisphere for the first time.

    Uranus is sometimes called the 'sideways' planet, because its rotation axis tipped more than 90 degrees from the planet's orbit around the Sun. The 'year' on Uranus lasts 84 Earth years, which creates extremely long seasons - winter in the northern hemisphere has lasted for nearly 20 years. Uranus has also been called bland and boring, because no clouds have been detectable in ground-based images of the planet. Even to the cameras of the Voyager spacecraft in 1986, Uranus presented a nearly uniform blank disk, and discrete clouds were detectable only in the southern hemisphere. Voyager flew over the planet's cloud tops near the dead of northern winter (when the northern hemisphere was completely shrouded in darkness).

    Spring has finally come to the northern hemisphere of Uranus. The newest images, both the visible-wavelength ones described here and those taken a few days earlier with the Near Infrared and Multi-Object Spectrometer (NICMOS) by Erich Karkoschka (University of Arizona), show a planet with banded structure and detectable clouds.

    Two images are shown here. The 'aqua' image (on the left) is taken at 5,470 Angstroms, which is near the human eye's peak response to wavelength. Color has been added to the image to show what a person on a spacecraft near Uranus might see. Little structure is evident at this wavelength, though with image-processing techniques, a small cloud can be seen near the planet's northern limb

  19. HUBBLE SPOTS NORTHERN HEMISPHERIC CLOUDS ON URANUS

    NASA Technical Reports Server (NTRS)

    2002-01-01

    Using visible light, astronomers for the first time this century have detected clouds in the northern hemisphere of Uranus. The newest images, taken July 31 and Aug. 1, 1997 with NASA Hubble Space Telescope's Wide Field and Planetary Camera 2, show banded structure and multiple clouds. Using these images, Dr. Heidi Hammel (Massachusetts Institute of Technology) and colleagues Wes Lockwood (Lowell Observatory) and Kathy Rages (NASA Ames Research Center) plan to measure the wind speeds in the northern hemisphere for the first time. Uranus is sometimes called the 'sideways' planet, because its rotation axis is tipped more than 90 degrees from the planet's orbit around the Sun. The 'year' on Uranus lasts 84 Earth years, which creates extremely long seasons - winter in the northern hemisphere has lasted for nearly 20 years. Uranus has also been called bland and boring, because no clouds have been detectable in ground-based images of the planet. Even to the cameras of the Voyager spacecraft in 1986, Uranus presented a nearly uniform blank disk, and discrete clouds were detectable only in the southern hemisphere. Voyager flew over the planet's cloud tops near the dead of northern winter (when the northern hemisphere was completely shrouded in darkness). Spring has finally come to the northern hemisphere of Uranus. The newest images, both the visible-wavelength ones described here and those taken a few days earlier with the Near Infrared and Multi-Object Spectrometer (NICMOS) by Erich Karkoschka (University of Arizona), show a planet with banded structure and detectable clouds. Two images are shown here. The 'aqua' image (on the left) is taken at 5,470 Angstroms, which is near the human eye's peak response to wavelength. Color has been added to the image to show what a person on a spacecraft near Uranus might see. Little structure is evident at this wavelength, though with image-processing techniques, a small cloud can be seen near the planet's northern limb (rightmost

  20. Comment on 'Are survival rates for northern spotted owls biased?'

    USGS Publications Warehouse

    Franklin, A.B.; Nichols, J.D.; Anthony, R.G.; Burnham, K.P.; White, Gary C.; Forsman, E.D.; Anderson, D.R.

    2006-01-01

    Loehle et al. recently estimated survival rates from radio-telemetered northern spotted owls (Strix occidentalis caurina (Merriam, 1898)) and suggested that survival rates estimated for this species from capture-recapture studies were negatively biased, which subsequently resulted in the negatively biased estimates of rates of population change (lambda) reported by Anthony et al. (Wildl. Monogr. No. 163, pp. 1-47 (2006)). We argue that their survival estimates were inappropriate for comparison with capture-recapture estimates because (i) the manner in which they censored radio-telemetered individuals had the potential to positively bias their survival estimates, (ii) their estimates of survival were not valid for evaluating bias, and (iii) the size and distribution of their radiotelemetry study areas were sufficiently different from capture-recapture study areas to preclude comparisons. In addition, their inferences of negative bias in rates of population change estimated by Anthony et al. were incorrect and reflected a misunderstanding about those estimators.