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Sample records for activity western blotting

  1. Western blots.

    PubMed

    Freeman, Lita A

    2013-01-01

    Western analysis of apolipoproteins, lipoproteins, and proteins involved in lipoprotein metabolism can be challenging due to their size, hydrophobic nature, and, in some cases, low abundance. Here we describe a Western blotting method that has been used successfully for many proteins involved in lipoprotein metabolism, as well as intact LDL or HDL particles. Proteins or lipoprotein particles separated by gel electrophoresis are transferred to a PVDF membrane in a Hoefer TE22 transfer tank with Tris-Glycine-SDS-Methanol transfer buffer. The membrane is blocked with 3 % BSA/5 % milk to prevent nonspecific binding of antibody to the membrane and is then incubated with primary antibody that binds specifically to the protein of interest. After washing away unbound primary antibody, the membrane is then incubated with an HRP-labeled secondary antibody that binds primary antibody. After washing away unbound secondary antibody, the membrane is then incubated with a substrate for HRP, generating a chemiluminescent signal at the location of the protein of interest. The protein is visualized by exposing the membrane to an autoradiography film or an imaging device. Information on the use of several human antibodies, including apoA-I, A-II, apoB, apoC-II, apoC-III, apoD, apoL1, apoM, PON1, SAA, ABCA1, nitrotyrosine, and LCAT, is provided. This method can be used for Western blotting of virtually any protein as well as native lipoprotein particles. PMID:23912997

  2. 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...

  3. 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

  4. 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

  5. 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

  6. 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

  7. Studying protein-protein interactions via blot overlay/far western blot.

    PubMed

    Hall, Randy A

    2015-01-01

    Blot overlay is a useful method for studying protein-protein interactions. This technique involves fractionating proteins on SDS-PAGE, blotting to nitrocellulose or PVDF membrane, and then incubating with a probe of interest. The probe is typically a protein that is radiolabeled, biotinylated, or simply visualized with a specific antibody. When the probe is visualized via antibody detection, this technique is often referred to as "Far Western blot." Many different kinds of protein-protein interactions can be studied via blot overlay, and the method is applicable to screens for unknown protein-protein interactions as well as to the detailed characterization of known interactions.

  8. 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

  9. 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.

  10. Using a large area CMOS APS for direct chemiluminescence detection in Western blotting electrophoresis

    NASA Astrophysics Data System (ADS)

    Esposito, Michela; Newcombe, Jane; Anaxagoras, Thalis; Allinson, Nigel M.; Wells, Kevin

    2012-03-01

    Western blotting electrophoretic sequencing is an analytical technique widely used in Functional Proteomics to detect, recognize and quantify specific labelled proteins in biological samples. A commonly used label for western blotting is Enhanced ChemiLuminescence (ECL) reagents based on fluorescent light emission of Luminol at 425nm. Film emulsion is the conventional detection medium, but is characterized by non-linear response and limited dynamic range. Several western blotting digital imaging systems have being developed, mainly based on the use of cooled Charge Coupled Devices (CCDs) and single avalanche diodes that address these issues. Even so these systems present key drawbacks, such as a low frame rate and require operation at low temperature. Direct optical detection using Complementary Metal Oxide Semiconductor (CMOS) Active Pixel Sensors (APS)could represent a suitable digital alternative for this application. In this paper the authors demonstrate the viability of direct chemiluminescent light detection in western blotting electrophoresis using a CMOS APS at room temperature. Furthermore, in recent years, improvements in fabrication techniques have made available reliable processes for very large imagers, which can be now scaled up to wafer size, allowing direct contact imaging of full size western blotting samples. We propose using a novel wafer scale APS (12.8 cm×13.2 cm), with an array architecture using two different pixel geometries that can deliver an inherently low noise and high dynamic range image at the same time representing a dramatic improvement with respect to the current western blotting imaging systems.

  11. A high-affinity reversible protein stain for Western blots.

    PubMed

    Antharavally, Babu S; Carter, Brad; Bell, Peter A; Krishna Mallia, A

    2004-06-15

    We describe a reversible staining technique, using MemCode, a reversible protein stain by which proteins can be visualized on nitrocellulose and polyvinylidine fluoride (PVDF) membranes without being permanently fixed to the membrane itself. This allows subsequent immunoblot analysis of the proteins to be performed. The procedure is applicable only to protein blots on nitrocellulose and PVDF membranes. MemCode is a reversible protein stain composed of copper as a part of an organic complex that interacts noncovalently with proteins. MemCode shows rapid protein staining, taking 30s to 1 min for completion. The method is simple and utilizes convenient application conditions that are compatible with the matrix materials and the protein. The stain is more sensitive than any previously described dye-based universal protein staining system. The turquoise-blue-stained protein bands do not fade with time and are easy to photograph compared to those stained with Ponceau S. Absorbance in the blue region of the spectrum offers good properties for photo documentation and avoids interference from common biological chromophores. The stain on the protein is easily reversible in 2 min for nitrocellulose membrane and in 10 min for PVDF membrane with MemCode stain eraser. The stain is compatible with general Western blot detection systems, and membrane treatment with MemCode stain does not interfere with conventional chemiluminescent or chromogenic detection using horseradish peroxide and alkaline phosphatase substrates. The stain is also compatible with N-terminal sequence analysis of proteins.

  12. 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

  13. Western blotting by thin-film direct coating.

    PubMed

    Yen, Yi-Kuang; Jiang, Yi-Wei; Chang, Shih-Chung; Wang, An-Bang

    2014-05-20

    A novel thin-film direct coating (TDC) technique was developed to markedly reduce the amount of antibody required for Western blotting (WB). Automatic application of the technique for a few seconds easily and homogeneously coats the specific primary antibody on the polyvinylidene fluoride (PVDF) membrane. While conventional WB requires 0.4 μg of the primary antibody, the proposed technique only uses 4 × 10(-2) μg, which can be reduced further to 4 × 10(-5) μg by reducing the coater width. Moreover, the proposed process reduces antibody probing times from 60 to 10 min. The quantification capability of TDC WB showed high linearity within a 4-log2 dynamic range for detecting target antigen glutathione-S-transferase. Furthermore, TDC WB can specifically detect the extrinsic glutathione-S-transferase added in the Escherichia coli or 293T cell lysate with better staining sensitivity than conventional WB. TDC WB can also clearly probe the intrinsic β-actin, α-tubulin, and glyceraldehyde 3-phosphate dehydrogenase, which are usually used as control proteins in biological experiments. This novel technique has been shown to not only have valuable potential for increasing WB efficiency but also for providing significant material savings for future biomedical applications. PMID:24773468

  14. Multicenter evaluation of the novel ABN Western blot (immunoblot) system in comparison with an enzyme-linked immunosorbent assay and a different Western blot.

    PubMed

    Weber, B; Hess, G; Enzensberger, R; Harms, F; Evans, C J; Hamann, A; Doerr, H W

    1992-03-01

    A new, modular Western blot (immunoblot) system for human immunodeficiency virus (HIV) antibodies (ABN WesPage; Wellcome) was compared with enzyme immunoassays (Wellcome, Behringwerke, and Abbott) and with a U.S. Food and Drug Administration (FDA)-licensed Western blot (DuPont) in a multicenter study. A total of 649 serum samples from HIV patients at different stages of the disease, as well as from high-risk patients, from patients with conditions unrelated to AIDS, and from healthy blood donors, were used in the evaluation along with nine seroconversion panels. For evaluation of Western blot reactivity, both Centers for Disease Control (CDC) and FDA criteria were used. With the DuPont Western blot as the reference assay, the overall sensitivity and specificity of the ABN WesPage were 100 and 99.1%, respectively, when indeterminate results were not taken into account and when both tests were interpreted in accordance with CDC criteria. The DuPont Western blot detected significantly more antibodies to pol and gag gene products than the ABN WesPage. The ABN WesPage showed a higher positive rate of detection of viral envelope band gp160. When both Western blots were interpreted in accordance with CDC criteria, the ABN WesPage and the DuPont Western blot yielded 9.3 and 10.4% indeterminate results, respectively. When the DuPont Western blot was interpreted in accordance with the manufacturer's instructions (FDA criteria), 25.7% of the samples tested were regarded as indeterminate. The choice of interpretation criteria is of paramount importance for the evaluation of HIV Western blot patterns.

  15. 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

  16. 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…

  17. Luminex xMAP combined with Western blot improves HIV diagnostic sensitivity.

    PubMed

    Kong, Weiwei; Li, Yan; Cheng, Shaohui; Yan, Chen; An, Shiping; Dong, Zheng; Yan, Lina; Yuan, Yuhua

    2016-01-01

    Currently, Western blot is used to confirm the initial serodiagnosis of HIV infection by antibody detection. However, a major deficiency of the Western blot relates to a lack of sufficient sensitivity in detecting HIV antibodies. This report describes a simple, sensitive and inexpensive bead-based assay for detection of early HIV infection. A panel of 138 positive specimens including 105 blood donors and 33 MSM with known Western blot results were evaluated using Luminex xMAP at Tianjin Centers for Disease Control and Prevention (CDC). We demonstrate a superior sensitivity of Luminex xMAP compared with Western blot. Of the 87 confirmed HIV positive blood donors, Western blot only confirmed 65 cases with 74.7% (65/87) sensitivity while Luminex xMAP identified 72 cases with 82.8% (72/87) sensitivity (p<0.05). Western blot and Luminex xMAP verified 13 and 19 of 33 MSM specimens, respectively. The sensitivity was 39.4% (13/33) for Western blot and 57.6% (19/33) for Luminex xMAP (p<0.1). Luminex xMAP combined with Western blot improves the diagnostic sensitivity of HIV infection at an early stage, and reduces the chances of missed diagnosis.

  18. Automated capillary Western dot blot method for the identity of a 15-valent pneumococcal conjugate vaccine.

    PubMed

    Hamm, Melissa; Ha, Sha; Rustandi, Richard R

    2015-06-01

    Simple Western is a new technology that allows for the separation, blotting, and detection of proteins similar to a traditional Western except in a capillary format. Traditionally, identity assays for biological products are performed using either an enzyme-linked immunosorbent assay (ELISA) or a manual dot blot Western. Both techniques are usually very tedious, labor-intensive, and complicated for multivalent vaccines, and they can be difficult to transfer to other laboratories. An advantage this capillary Western technique has over the traditional manual dot blot Western method is the speed and the automation of electrophoresis separation, blotting, and detection steps performed in 96 capillaries. This article describes details of the development of an automated identity assay for a 15-valent pneumococcal conjugate vaccine, PCV15-CRM197, using capillary Western technology.

  19. Single-cell Western blotting after whole-cell imaging to assess cancer chemotherapeutic response.

    PubMed

    Kang, Chi-Chih; Lin, Jung-Ming G; Xu, Zhuchen; Kumar, Sanjay; Herr, Amy E

    2014-10-21

    Intratumor heterogeneity remains a major obstacle to effective cancer therapy and personalized medicine. Current understanding points to differential therapeutic response among subpopulations of tumor cells as a key challenge to successful treatment. To advance our understanding of how this heterogeneity is reflected in cell-to-cell variations in chemosensitivity and expression of drug-resistance proteins, we optimize and apply a new targeted proteomics modality, single-cell western blotting (scWestern), to a human glioblastoma cell line. To acquire both phenotypic and proteomic data on the same, single glioblastoma cells, we integrate high-content imaging prior to the scWestern assays. The scWestern technique supports thousands of concurrent single-cell western blots, with each assay comprised of chemical lysis of single cells seated in microwells, protein electrophoresis from those microwells into a supporting polyacrylamide (PA) gel layer, and in-gel antibody probing. We systematically optimize chemical lysis and subsequent polyacrylamide gel electrophoresis (PAGE) of the single-cell lysate. The scWestern slides are stored for months then reprobed, thus allowing archiving and later analysis as relevant to sparingly limited, longitudinal cell specimens. Imaging and scWestern analysis of single glioblastoma cells dosed with the chemotherapeutic daunomycin showed both apoptotic (cleaved caspase 8- and annexin V-positive) and living cells. Intriguingly, living glioblastoma subpopulations show up-regulation of a multidrug resistant protein, P-glycoprotein (P-gp), suggesting an active drug efflux pump as a potential mechanism of drug resistance. Accordingly, linking of phenotype with targeted protein analysis with single-cell resolution may advance our understanding of drug response in inherently heterogeneous cell populations, such as those anticipated in tumors.

  20. [Analysis of new criteria for increasing the specificity of commercial Western blots].

    PubMed

    Gershy-Damet, G M; Koffi, K; Soro, B; Sanon, S; Traore, M; Assoa, A; N'Goran, K; N'Gom, A

    1992-01-01

    We examined the frequency of serum cross-reactivity on Western blot for HIV1 and HIV2. 661 patients with tuberculosis in Abidjan, and 4,899 asymptomatic persons for HIV1 and HIV2 infections were tested. All specimens positive on ELISA for HIV1 or HIV2 were further characterized by synthetic peptide based tests. Confirmed positive samples were tested by HIV1 and HIV2 specific Western blot criteres utilisis. Dual serologic reactivity on synthetic peptide tests was significantly more frequent in HIV positive patients with tuberculosis than asymptomatic subjects. Positive HIV1 Western blots were seen in 61%-86% of specimens positive for HIV2 only on synthetic peptide tests. [Cross-reactivity, to HIV2 Western blots by HIV1 positive specimens was significantly more frequent in patients with tuberculosis than in asymptomatic subjects.] Using recently recommended criteria for HIV1 and HIV2 Western blot interpretation (presence of 2 env bands) reduced the overall proportion of HIV1 positive specimens having a positive HIV2 Western blot from 39% to 14% and HIV2 positive specimens having a positive HIV1 Western blot from 31% to 8%.

  1. Western blotting is useful in the salivary diagnosis of Helicobacter pylori infection

    PubMed Central

    Ballam, L; Mendall, M; Asante, M; Morris, J; Strachan, D; Whincup, P; Cook, D

    2000-01-01

    Background—The salivary diagnosis of Helicobacter pylori infection offers attractive possibilities for the epidemiological study of infection in children. Salivary enzyme linked immunosorbent assay (ELISA) is less reliable then serum ELISA, owing to variable transudation of immunoglobulin. In addition, children are more difficult to study because of lower specific serum antibody concentrations to H pylori. The performance of salivary western blotting in comparison with serum western blotting and serum ELISA was investigated in school children. Subjects and methods—Paired serum and saliva specimens were obtained from 665 school children aged 9–11 in 10 British towns. All saliva and serum specimens were first analysed by ELISA; subsequently, western blotting of both specimens was performed on 31 and 34 specimens, respectively, to establish the criteria for positivity for western blotting. The remaining 121 specimens were then tested blindly and saliva was compared with the serum. Results—The sensitivity and specificity of salivary ELISA in the 665 specimens was 32 of 50 (64%) and 530 of 691 (87%), respectively, when compared with serum ELISA. The western blotting validation was performed on 28 subjects with positive serum and positive salivary ELISA, 28 saliva positives with negative serum, 16 saliva negatives with positive serum, and 50 doubly negative subjects. Compared with serum western blots, the sensitivity and specificity of salivary western blots was 38 of 47 (81%) and 68 of 75 (91%), respectively. Using serum ELISA as the gold standard, the sensitivity and specificity were 32 of 44 (73%) and 72 of 78 (92%), respectively, the specificity being significantly higher than salivary ELISA (p < 0.001). Conclusion—Salivary western blotting for IgG is useful in the diagnosis of H pylori infection and is superior to ELISA. It also permits the identification of pathogenic strains. Key Words: Helicobacter pylori • western blotting • enzyme linked

  2. SDS-Polyacrylamide Electrophoresis and Western Blotting Applied to the Study of Asthma.

    PubMed

    García-Solaesa, Virginia; Abad, Sara Ciria

    2016-01-01

    Western blotting is used to analyze proteins after being separated by electrophoresis and subsequently electro-transferred to a membrane. Once immobilized, a specific protein can be identified through its reaction with a labeled antibody or antigen. It is a methodology commonly used in biomedical research such as asthma studies, to assess the pathways of inflammatory mediators involved in the disease.Here, we describe an example of western blotting to determine the factors involved in asthma. In this chapter, the methodology of western blotting is reviewed, paying attention on potential problems and giving interesting recommendations. PMID:27300534

  3. Microfluidic integration of Western blotting is enabled by electrotransfer-assisted sodium dodecyl sulfate dilution.

    PubMed

    Hou, Chenlu; Herr, Amy E

    2013-01-01

    We integrate sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) with subsequent antibody probing in a single, monolithic microdevice to realize microfluidic Western blotting. A hurdle to successful on-chip Western blotting lies in restoring antibody recognition of previously sized (denatured, reduced) proteins. To surmount this hurdle, we locally dilute free SDS from SDS-protein complexes using differential electromigration of the species during electrotransfer between SDS-PAGE and blotting regions of a microchamber. Local dilution of SDS minimizes re-association of SDS with proteins offering means to restore antibody binding affinity to proteins after SDS-PAGE. To achieve automated, programmable operation in a single instrument, we utilize a 1 × 2 mm(2) glass microchamber photopatterned with spatially distinct, contiguous polyacrylamide regions for SDS-PAGE, electrotransfer, and antibody blotting. Optimization of both the SDS-PAGE and electrotransfer conditions yields transfer distances of <1 mm (40 s). The Western blot is completed in 180 s, with fully automated assay operation using programmable voltage control. After SDS-PAGE and electrotransfer, we observe ~80% capture of protein band mass on the blotting region for a model protein, C-reactive protein. This novel microfluidic Western blot approach introduces fine transport control for in-transit protein handling to form the basis for an automated, rapid alternative to conventional slab-gel Western blotting. PMID:23042290

  4. Detection and quantification of protein-protein interactions by far-western blotting.

    PubMed

    Jadwin, Joshua A; Mayer, Bruce J; Machida, Kazuya

    2015-01-01

    Far-western blotting is a convenient method to characterize protein-protein interactions, in which protein samples of interest are immobilized on a membrane and then probed with a non-antibody protein. In contrast to western blotting, which uses specific antibodies to detect target proteins, far-western blotting detects proteins on the basis of the presence or absence of binding sites for the protein probe. When specific modular protein binding domains are used as probes, this approach allows characterization of protein-protein interactions involved in biological processes such as signal transduction, including interactions regulated by posttranslational modification. We here describe a rapid and simple protocol for far-western blotting, in which GST-tagged Src homology 2 (SH2) domains are used to probe cellular proteins in a phosphorylation-dependent manner. We also present a batch quantification method that allows for the direct comparison of probe binding patterns.

  5. Characterization of a biopharmaceutical protein and evaluation of its purification process using automated capillary Western blot.

    PubMed

    Xu, Dong; Mane, Sarthak; Sosic, Zoran

    2015-01-01

    This paper describes the application of an automated size-based capillary Western blot system (Sally instrument) from ProteinSimple, Inc., for biopharmaceutical fusion-Fc protein characterization and evaluation of its purification process. The fusion-Fc protein column purification from an excess of single chain Fc polypeptide and removal of an enzyme coexpressed for protein maturation have been demonstrated using an automated capillary Western system. The clearance of a selected host cell protein (HCP) present in cell culture of fusion-Fc protein was also quantitatively monitored throughout the protein purification process. Additionally, the low levels of fusion-Fc product-related impurities detected by traditional slab gel Western blot were confirmed by the automated capillary Western system. Compared to the manual approach, the automated capillary Western blot provides the advantages of ease of operation, higher sample throughput, greater linearity range, and higher precision for protein quantitation.

  6. 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

  7. Charge standards as reference points in two-dimensional western blot patterns

    SciTech Connect

    Zhang, J.S.; Tollaksen, S.L.; Giometti, C.S.

    1986-01-01

    High-resolution two-dimensional electrophoresis can be used to separate hundreds or even thousands of proteins from complex mixtures. Specific proteins in 2DE patterns can be identified by using Western Blotting. Correlation between a single stained spot on a nitrocellulose blot and the same spot in a stained 2DE gel pattern can be difficult since no reference points are available on the blot. Inclusion of a protein or proteins to serve as reference or standard points in both the blotted pattern and the stained gel pattern would make pattern recognition much more rapid and reliable. In this paper the use of a commercial charge standard (carbamylated creatine phosphokinase mix) was used to produce reference points in both gel patterns and corresponding nitrocellulose blots to simplify recognition of individual protein spots in complicated gel patterns. 8 refs., 2 figs.

  8. Investigation of immunofluorescence cross-reactions against Trichinella spiralis by western blot (immunoblot) analysis.

    PubMed Central

    Robert, F; Weil, B; Kassis, N; Dupouy-Camet, J

    1996-01-01

    Immunofluorescence cross-reactions in Trichinella spiralis serodiagnosis are sometimes difficult to identify. We compared the results of an indirect immunofluorescence assay and the profiles obtained by Western blot (immunoblot) analysis for three groups of patients: 10 T. spiralis-infected patients, 10 patients with autoimmune diseases, and 7 patients with parasitic diseases other than trichinellosis. The degree of immunofluorescence cross-reaction was variable. Western blotting allowed us to differentiate Trichinella infection from other parasitic diseases. In 3 of 10 serum samples from patients with autoimmune diseases, bands which had the same sizes as Trichinella bands were observed, and they could correspond to shared epitopes such as heat shock proteins. PMID:8877138

  9. 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

  10. 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…

  11. 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

  12. Post-obstruction rat sperm autoantigens identified by two-dimensional gel electrophoresis and western blotting.

    PubMed

    Flickinger, C J; Bush, L A; Williams, M V; Naaby-Hansen, S; Howards, S S; Herr, J C

    1999-05-01

    Although antisperm autoantibody responses to obstruction of the male reproductive system have been documented, information on the nature of the cognate sperm autoantigens has been limited. In the present study, the patterns of sperm autoantigens recognized by sera from rats after obstruction of the vas deferens or epididymis were studied by high resolution two-dimensional (2-D) gel electrophoresis and western blotting. Comparisons of patterns of autoantigens stained on 2-D western blots of sera from prepubertal vasectomy, prepubertal epididymal ligation and adult vasectomy groups revealed both similarities and differences. Sera from sham-operated animals showed no detectable reaction or much lighter staining of a small number of spots. Visualization of sperm autoantigens on 2-D western blots supported the hypothesis that there is a relatively small set of sperm proteins that can be regarded as dominant post-obstruction sperm autoantigens because they are recognized by multiple post-obstruction sera. The 2-D analysis revealed previously undetected distinctions in the autoantigens recognized after adult and prepubertal vasectomy, as well as variations with the site of obstruction. These differences in the response may be due in part to changes in antigens of spermatozoa in different parts of the tract and at different ages, as well as variations in exposure of sperm cell proteins to the immune system resulting from the sites of spermatic granulomas. Preparative 2-D gels and western blotting with post-obstruction sera are now being used to identify specific sperm autoantigens by microsequencing of selected proteins. PMID:10392780

  13. 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

  14. 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...

  15. Intermittent microwave irradiation facilitates antigen-antibody reaction in Western blot analysis.

    PubMed

    Liu, Yu-Ting; Toyokuni, Shinya

    2009-01-01

    We established a shortened protocol for western blot analysis using intermittent microwave irradiation. With this method, the procedure is completed within 1 h after applying the primary antibody, and thus greatly saves time. This procedure appears to be applicable to any antibody based on our experience of several years.

  16. 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…

  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. A streamlined Western blot exercise: An efficient and greener approach in the laboratory classroom.

    PubMed

    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 from staining and destaining, whereas with western blotting it is the times required for antibody incubations and the numerous wash steps. This laboratory exercise incorporates 2,2,2-trichloroethanol (TCE) into the SDS-PAGE gel allowing for visualization of migrated proteins in a matter of minutes, saving both the time and chemical waste associated with traditional Coomassie staining. Additionally, TCE staining does not affect protein transfer eliminating the requirement for duplicated gels for total protein and western analyses. Protein transfer can be confirmed immediately without the use of Ponceau S staining. Lastly, this western blot procedure has been further shortened by using an HRP-conjugated primary antibody, which eliminates the secondary antibody incubation and washes, and uses a colorimetric detection to allow for visualization by students without the need for specialized equipment. PMID:26153352

  19. Nanogold Immunodetection Detection Systems for the Identification of Autoantigens by Western Blotting.

    PubMed

    Moore, Jacen S; Scofield, R Hal

    2015-01-01

    Nanogold-conjugated immunodetection systems are now widely and commercially available for use in a number of research applications including electron microscopy, light microscopy, and western blotting. Nanogold clusters are small, uniform in size, and stable, unlike gold colloids historically used in protein detection. Covalent linkage of nanogold particles to secondary antibodies prevents dissociation of the gold particles during the staining process, making protein detection reliable, antigen specific, and highly sensitive. Nanogold labeling is extremely versatile and can be used in conjunction with other staining methodologies including Alexa Fluor immunofluorescence detection to perform coupled staining procedures. Silver enhancement increases the limits of sensitivity for nanogold staining, thus improving detection signals for antigens with reduced expression levels. Herein, we describe the use of nanogold-silver detection as an immunodetection system for standard western blotting of autoantigens.

  20. 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

  1. Use of a Western blot technique for the serodiagnosis of glanders

    PubMed Central

    2011-01-01

    Background The in vivo diagnosis of glanders relies on the highly sensitive complement fixation test (CFT). Frequently observed false positive results are troublesome for veterinary authorities and cause financial losses to animal owners. Consequently, there is an urgent need to develop a test with high specificity. Hence, a Western blot assay making use of a partly purified lipopolysaccaride (LPS) containing antigen of three Burkholderia mallei strains was developed. The test was validated investigating a comprehensive set of positive and negative sera obtained from horses and mules from endemic and non endemic areas. Results The developed Western blot assay showed a markedly higher diagnostic specificity when compared to the prescribed CFT and therefore can be used as a confirmatory test. However, the CFT remains the test of choice for routine testing of glanders due to its high sensitivity, its feasibility using standard laboratory equipment and its worldwide distribution in diagnostic laboratories. Conclusions The CFT should be amended by the newly validated Western blot to increase the positive likelihood ratio of glanders serodiagnosis in non endemic areas or areas with low glanders prevalence. Its use for international trade of horses and mules should be implemented by the OIE. PMID:21247488

  2. 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

  3. Detection of IgA and IgM antibodies to HIV-1 in neonates by radioimmune western blotting.

    PubMed Central

    Portincasa, P.; Conti, G.; Re, M. C.; Chezzi, C.

    1992-01-01

    OBJECTIVE--To detect infection with HIV-1 by IgA and IgM response at birth in children born to HIV-1 seropositive mothers. DESIGN--Western blotting and radioimmune western blotting on stored sera from infected and uninfected babies born to HIV-1 seropositive mothers. Sera were pretreated to remove IgG. SETTING--Parma and Bologna, Italy. SUBJECTS--12 infected and five uninfected babies born to HIV-1 seropositive mothers and three babies born to seronegative mothers. MAIN OUTCOME MEASURES--Effectiveness of western blotting and radioimmune western blotting in detecting antibodies to HIV-1 gene products. RESULTS--With conventional western blotting we found IgA class antibodies to HIV-1 proteins in serum from three out of 12 infected children; in two of these three the serum was collected at age 3 months (positive controls). Radioimmune western blotting detected both IgA and IgM antibodies in serum from all infected children tested, whereas all serum from uninfected children born to seropositive and seronegative mothers showed no such antibodies. CONCLUSION--Although the technique should be tested on more patients, radioimmune western blotting seems to be a valuable tool for serological diagnosis of congenital HIV-1 infection at birth in neonates born to seropositive mothers. Images FIG 1 FIG 2 PMID:1628052

  4. Evaluating dot and Western blots using image analysis and pixel quantification of electronic images.

    PubMed

    Vierck, J L; Bryne, K M; Dodson, M V

    2000-01-01

    Inexpensive computer imaging technology was used to assess levels of insulin-like growth factor-I (IGF-I) on dot blots (DB) and alpha-Actinin on Western blots (WB). In the first procedure, known IGF-I samples were dotted on nitrocellulose membranes using a vacuum manifold. After the DB were developed and dried, the images were digitized using an HP Deskscan II flat bed scanner, exported into Image-Pro Plus and analyzed by taking the combined mean of 45 degrees and 135 degrees sample lines drawn through each dot. Dot blots corresponding to a linear concentration range from 10 to 300 ng IGF-I were assessed by this method. In the second procedure, WB were scanned with a ScanJet 3c flat bed scanner and their backgrounds were clarified using Image-Pro Plus. A second image analysis program, Alpha Imager 2000, was then used to define the boundaries of protein bands, assess pixel number and density, and to obtain final numerical data for quantifying alpha-Actinin on the WB. Collectively, the results of these two studies suggest that specific proteins may be evaluated by using relatively inexpensive image analysis software systems via pixel quantification of electronic images. PMID:11549944

  5. 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

  6. Analyzing the Homeostasis of Signaling Proteins by a Combination of Western Blot and Fluorescence Correlation Spectroscopy

    PubMed Central

    Chung, Yi-Da; Sinzinger, Michael D.; Bovee-Geurts, Petra; Krause, Marina; Dinkla, Sip; Joosten, Irma; Koopman, Werner J.; Adjobo-Hermans, Merel J.W.; Brock, Roland

    2011-01-01

    The determination of intracellular protein concentrations is a prerequisite for understanding protein interaction networks in systems biology. Today, protein quantification is based either on mass spectrometry, which requires large cell numbers and sophisticated measurement protocols, or on quantitative Western blotting, which requires the expression and purification of a recombinant protein as a reference. Here, we present a method that uses a transiently expressed fluorescent fusion protein of the protein-of-interest as an easily accessible reference in small volumes of crude cell lysates. The concentration of the fusion protein is determined by fluorescence correlation spectroscopy, and this concentration is used to calibrate the intensity of bands on a Western blot. We applied this method to address cellular protein homeostasis by determining the concentrations of the plasma membrane-located transmembrane scaffolding protein LAT and soluble signaling proteins in naïve T cells and transformed T-cell lymphoma (Jurkat) cells (with the latter having nine times the volume of the former). Strikingly, the protein numbers of soluble proteins scaled with the cell volume, whereas that of the transmembrane protein LAT scaled with the membrane surface. This leads to significantly different stoichiometries of signaling proteins in transformed and naïve cells in concentration ranges that may translate directly into differences in complex formation. PMID:22261070

  7. 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

  8. Validation of endothelin B receptor antibodies reveals two distinct receptor-related bands on Western blot.

    PubMed

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

    2015-01-01

    Antibodies are important tools for the study of protein expression but are often used without full validation. In this study, we used Western blots to characterize antibodies targeted to the N or C terminal (NT or CT, respectively) and the second or third intracellular loop (IL2 or IL3, respectively) 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 50-kDa 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-kDa band but failed to detect endogenous ETB in rat brain. Bands detected by the CT- or IL3-targeted antibody were found to be unrelated to ETB. Our findings show that functional ETB can be detected at 50 or 37kDa on Western blot, with drastic differences in antibody affinity for these bands. The 37-kDa band likely reflects ETB processing, which appears to be dependent on cell type and/or culture condition.

  9. 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.

  10. HIV‑2 antibody detection after indeterminate or negative HIV‑1 Western blot in Cuba, 2005-2008.

    PubMed

    Díaz, Dervel F; Ortiz, Eva; Martín, Dayamí; Nibot, Carmen; Rizo, Adis; Silva, Eladio

    2012-01-01

    INTRODUCTION Differentiating between HIV-1 and HIV-2 infection is the first step to understanding HIV transmission, epidemiology and pathogenesis in geographical areas where both viruses circulate. In Cuba, positive results in mixed HIV-1/2 screening assays are confirmed by HIV-1 Western blot. Indeterminate results constitute the main limitation of this test and HIV-2 infection is among their possible causes; hence the importance of second-stage screening and confirmatory tests for HIV-2 infection. OBJECTIVE Investigate the contribution of HIV-2 antibodies to negative or indeterminate HIV-1 Western blot results in serum samples from 2005 through 2008 in Cuba. METHODS HIV-2 reactivity was studied using the ELISA DAVIH-VIH-2 diagnostic kit (Cuba) in 1723 serum samples with negative or indeterminate results for HIV-1 Western blot from January 2005 through December 2008. Duplicate sera reactive by ELISA were confirmed by HIV-2 Western blot, results interpreted according to WHO criteria. The epidemiological interview established by Cuba's National Program for Prevention and Control Sexually-Transmitted Diseases and HIV/AIDS was applied to HIV-2 Western blot-positive patients. RESULTS Among all sera studied, HIV-2 ELISA identified 12 reactive serum samples (0.70%) and 1711 non-reactive (99.30%). Western blot analysis of the 12 ELISA-reactive samples confirmed two positive samples (16.67%), 4 negative (33.33%) and 6 indeterminate (50%). Positive samples reacted against the p16, p26, gp36, p53, p56, p68 and gp105 proteins. All 12 ELISA-reactive samples belonged to the HIV-1 Western blot indeterminate group. The two HIV-2-positive samples showed well defined reactivity to gp160, p53, p55 and p34 of HIV-1. HIV-1 seroconversion was observed in all 10 remaining samples during serological followup. CONCLUSIONS Two new HIV-2 seropositive cases were diagnosed using DAVIH-VIH-2 and HIV-2 Western blot in indeterminate HIV-1 Western blot samples. Results support the recommendation

  11. 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

  12. 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

  13. Detection of Hypoderma actaeon infestation in Cervus elaphus with ELISA and western blotting.

    PubMed

    Domínguez, Julia; Panadero, Rosario; de la Fuente-López, Concepción

    2010-07-01

    We used enzyme-linked immunosorbent assay (ELISA) and western blotting (WB) to evaluate the reactivity of first-stage larval extracts of Hypoderma lineatum with antibodies in sera from 76 red deer from an endemic area for Hypoderma actaeon. Antibodies in sera from deer infested with H. actaeon recognized hypodermin C from H. lineatum in both ELISA and WB assays. ELISA values were correlated with the epidemiology of the fly infestation in the area where the deer sera were collected. There was no clear relationship between anti-hypodermin C antibody levels and the age of the animals or the number of Hypoderma grubs found at necropsy in the hides of the deer. Our results suggest that first-instar antigens of H. lineatum can be used to diagnose natural infestations by H. actaeon by indicating the presence of first-stage larvae, which can help to accurately describe H. actaeon epidemiology. PMID:20688700

  14. Detection of Hypoderma actaeon infestation in Cervus elaphus with ELISA and western blotting.

    PubMed

    Domínguez, Julia; Panadero, Rosario; de la Fuente-López, Concepción

    2010-07-01

    We used enzyme-linked immunosorbent assay (ELISA) and western blotting (WB) to evaluate the reactivity of first-stage larval extracts of Hypoderma lineatum with antibodies in sera from 76 red deer from an endemic area for Hypoderma actaeon. Antibodies in sera from deer infested with H. actaeon recognized hypodermin C from H. lineatum in both ELISA and WB assays. ELISA values were correlated with the epidemiology of the fly infestation in the area where the deer sera were collected. There was no clear relationship between anti-hypodermin C antibody levels and the age of the animals or the number of Hypoderma grubs found at necropsy in the hides of the deer. Our results suggest that first-instar antigens of H. lineatum can be used to diagnose natural infestations by H. actaeon by indicating the presence of first-stage larvae, which can help to accurately describe H. actaeon epidemiology.

  15. 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

  16. Detection of yellowhead virus and Chinese baculovirus in penaeid shrimp by the Western blot technique.

    PubMed

    Nadala, E C; Tapay, L M; Cao, S; Loh, P C

    1997-12-01

    The continuing threat posed by viral diseases in cultured shrimp calls for the development of detection technologies for monitoring the animals, especially broodstock. Two of the most highly pathogenic viruses of penaeid shrimp are the yellow-head virus (YHV) and Chinese baculovirus (CBV, also called white spot baculovirus). A Western blot (WB) protocol capable of detecting YHV and CBV in the hemolymph of infected shrimp was developed. The use of the hemolymph as material for virus detection allowed for sample collection without sacrificing the animals. This protocol was highly specific, rapid, and sensitive enough to detect the presence of the viruses before the appearance of overt symptoms. It was also useful for demonstrating the growth of both viruses in primary shrimp lymphoid cell cultures.

  17. 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

  18. IgG-subclass-specific antibody reactivity to respiratory syncytial virus polypeptides investigated by western blot.

    PubMed

    Jankowski, M; Hornsleth, A; Olsen, P G

    1990-01-01

    IgG1 and IgG3 subclass-specific antibody reactivity (ABR) in serum samples obtained from infants and children in relation to acute lower respiratory disease caused by respiratory syncytial virus (RSV) infection was investigated by Western blot. IgG1 ABR was directed against the nucleocapsid polypeptides VPN and VPP as well as against the glycoproteins GP48 (F1) and GP90. IgG3 ABR was directed only against VPN and VPP. In infants, a low IgG1 reactivity against glycoproteins was observed. When serum samples obtained in the early convalescent phase were tested, ABR against GP48 and GP90 as well as against VPP differed with respect to RSV subtypes A and B. IgG1 ABR increased in the late convalescent phase, while IgG3 ABR decreased during this phase when serum samples from primary infections were tested.

  19. Study of anti-dengue NS1 antibody by western blot.

    PubMed

    Kuno, G; Vorndam, A V; Gubler, D J; Gómez, I

    1990-10-01

    The presence of anti-nonstructural protein (NS1) antibody in natural dengue infections has been suspected to be associated with development of dengue haemorrhagic fever (DHF). The Western blot technique was used to study the dynamics of the immune response to NS1 and to determine the frequency of anti-NS1 antibody among confirmed dengue patients in Indonesia, where DHF is common, and in Puerto Rico, where DHF is less frequently observed. Anti-NS1 antibody was rarely found in those with primary infections in either group. The antibody occurred at a significantly higher frequency in acute-phase serum samples from secondary infections in Indonesia than in those from Puerto Rico. No difference was observed, however, in Indonesian patients with secondary infection who had dengue fever or DHF.

  20. 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.

  1. Strategies for laboratory HIV testing: an examination of alternative approaches not requiring Western blot.

    PubMed Central

    Sato, P. A.; Maskill, W. J.; Tamashiro, H.; Heymann, D. L.

    1994-01-01

    Advances in laboratory tests for antibodies to human immunodeficiency virus (HIV) have permitted the development of alternative HIV testing strategies that do not require use of the Western blot approach. Three strategies are proposed. In strategy I, sera are tested for HIV antibody using an enzyme-linked immunosorbent assay (ELISA)/rapid/simple (ERS) test; in strategy II, sera reactive in an initial ERS test are retested using a second ERS test; strategy III involves retesting with a third ERS test all sera reactive in two previous ERS tests. Where the objective is identification of asymptomatic HIV-infected individuals, strategy III is proposed where HIV prevalences in the study population are < or = 10%, and strategy II at prevalences > 10%. Strategy II is recommended where the diagnosis of HIV-related disease requires HIV testing. For serosurveillance, strategy II is recommended if the prevalence is < or = 10%, and strategy I if the prevalences are > 10%. Use of strategy I is recommended for transfusion and transplantation safety, at any prevalence. Lower-cost laboratory HIV testing will permit such testing to become more widely available. PMID:8131248

  2. 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.

  3. Protein quantification by chemiluminescent Western blotting: elimination of the antibody factor by dilution series and calibration curve.

    PubMed

    Charette, Steve J; Lambert, Herman; Nadeau, Philippe J; Landry, Jacques

    2010-02-28

    Quantification of chemiluminescent signals from a Western blot is routinely used to determine the increase or the decrease in protein expression or modification in cell or tissue extracts. However, although scientists readily agree that such a procedure is not quantitative, it is nonetheless used quantitatively in most publications without appropriate controls that would increase the accuracy of the measurement. Here we reexamined this aspect and found that the primary antibody itself influences the relation between the Western blot signal and the protein amount on the membrane. This relation is non-linear and varies from one antibody to another. In that context, we strongly encourage researchers to use dilution series and calibration curve when quantifying protein by Western blot using chemiluminescent signal.

  4. Stain-Free total protein staining is a superior loading control to β-actin for Western blots.

    PubMed

    Gilda, Jennifer E; Gomes, Aldrin V

    2013-09-15

    Semi-quantification of proteins using Western blots typically involves normalization against housekeeping genes such as β-actin. More recently, Ponceau S and Coomassie blue staining have both been shown to be suitable alternatives to housekeeping genes as loading controls. Stain-Free total protein staining offers the advantage of no staining or destaining steps. Evaluation of the use of Stain-Free staining as an alternative to β-actin or the protein stain Ponceau S showed that Stain-Free staining was superior to β-actin and as good as or better than Ponceau S staining as a loading control for Western blots. PMID:23747530

  5. Evaluation of the Aspergillus Western Blot IgG Kit for Diagnosis of Chronic Aspergillosis

    PubMed Central

    Oliva, A.; Flori, P.; Hennequin, C.; Dubus, J.-C.; Reynaud-Gaubert, M.; Charpin, D.; Vergnon, J. M.; Gay, P.; Colly, A.; Piarroux, R.; Pelloux, H.

    2014-01-01

    Immunoprecipitin detection (IPD) is the current reference confirmatory technique for anti-Aspergillus antibody detection; however, the lack of standardization is a critical drawback of this assay. In this study, we evaluated the performance of the Aspergillus Western blot (Asp-WB) IgG kit (LDBio Diagnostics, Lyon, France), a recently commercialized immunoblot assay for the diagnosis of various clinical presentations of chronic aspergillosis. Three hundred eight serum samples from 158 patients with aspergillosis sensu lato (s.l.) were analyzed. More specifically, 267 serum samples were derived from patients with Aspergillus disease, including 89 cases of chronic pulmonary aspergillosis, 10 of aspergilloma, and 32 of allergic bronchopulmonary aspergillosis, while 41 samples were from patients with Aspergillus colonization, including 15 cystic fibrosis (CF) and 12 non-CF patients. For blood donor controls, the Asp-WB specificity was 94%, while the kit displayed a sensitivity for the aspergillosis s.l. diagnosis of 88.6%, with a diagnostic odds ratio (DOR) of 119 (95% confidence interval [CI], 57 to 251). The DOR values were 185.22 (95% CI,78.79 to 435.45) and 43.74 (95% CI, 15.65 to 122.20) for the diagnosis of Aspergillus disease and Aspergillus colonization, respectively. Among the patients, the sensitivities of the Asp-WB in the diagnosis of Aspergillus colonization were 100% and 41.7% in CF and non-CF patients, respectively. The Asp-WB yielded fewer false-negative results than did IPD. In conclusion, the Asp-WB kit performed well for the diagnosis of various clinical presentations of aspergillosis in nonimmunocompromised patients, with an enhanced standardization and a higher sensitivity than with IPD, which is the current reference method. PMID:25392351

  6. Evaluation of the Aspergillus Western blot IgG kit for diagnosis of chronic aspergillosis.

    PubMed

    Oliva, A; Flori, P; Hennequin, C; Dubus, J-C; Reynaud-Gaubert, M; Charpin, D; Vergnon, J M; Gay, P; Colly, A; Piarroux, R; Pelloux, H; Ranque, S

    2015-01-01

    Immunoprecipitin detection (IPD) is the current reference confirmatory technique for anti-Aspergillus antibody detection; however, the lack of standardization is a critical drawback of this assay. In this study, we evaluated the performance of the Aspergillus Western blot (Asp-WB) IgG kit (LDBio Diagnostics, Lyon, France), a recently commercialized immunoblot assay for the diagnosis of various clinical presentations of chronic aspergillosis. Three hundred eight serum samples from 158 patients with aspergillosis sensu lato (s.l.) were analyzed. More specifically, 267 serum samples were derived from patients with Aspergillus disease, including 89 cases of chronic pulmonary aspergillosis, 10 of aspergilloma, and 32 of allergic bronchopulmonary aspergillosis, while 41 samples were from patients with Aspergillus colonization, including 15 cystic fibrosis (CF) and 12 non-CF patients. For blood donor controls, the Asp-WB specificity was 94%, while the kit displayed a sensitivity for the aspergillosis s.l. diagnosis of 88.6%, with a diagnostic odds ratio (DOR) of 119 (95% confidence interval [CI], 57 to 251). The DOR values were 185.22 (95% CI,78.79 to 435.45) and 43.74 (95% CI, 15.65 to 122.20) for the diagnosis of Aspergillus disease and Aspergillus colonization, respectively. Among the patients, the sensitivities of the Asp-WB in the diagnosis of Aspergillus colonization were 100% and 41.7% in CF and non-CF patients, respectively. The Asp-WB yielded fewer false-negative results than did IPD. In conclusion, the Asp-WB kit performed well for the diagnosis of various clinical presentations of aspergillosis in nonimmunocompromised patients, with an enhanced standardization and a higher sensitivity than with IPD, which is the current reference method.

  7. 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

  8. MDR-TB Antibody Response (Western Blot) to Fractions of Isoniazid and Rifampicin Resistant Antigens of Mycobacterium tuberculosis.

    PubMed

    Hadizadeh Tasbiti, Alireza; Yari, Shamsi; Ghanei, Mostafa; Shokrgozar, Mohammad Ali; Bahrmand, Ahmadreza

    2015-12-01

    Drug-resistant TB poses a major threat to control of TB worldwide. Despite progress in the detection of Multidrug-resistant TB (MDR-TB) cases, a major diagnostic gap remains: 55% of reported TB patients estimated to have MDR-TB were not detected in 2013. MDR-TB antigens were conjugated to CNBr-activated Sepharose 4B. Specific polyclonal antibodies against MDR-TB Ags were prepared in rabbits using two boosted injections of the MDR-TB antigen. The antibodies were purified and treated with susceptible TB to remove any non-specific and cross-reactive antibodies. In the present study, comparative analysis of electrophoretic pattern of different antigens of INH/RIF-resistant TB were studied for identifying protein profiles. A RIF-resistant TB antigen was shown here to have different protein profiles from INH-resistant TB isolate. The results of Western blotting analysis showed that in the RIF- and INH-resistant antigenic fractions some bands of 14.4 and 45 kDa as immunogenic were common. Moreover, four bands of RIF-resistant TB antigen fractions (16, 19, 21, and 45 KDa) and one band of INH-resistant TB (about 26 KDa) were detected as diagnostic antigens. This study suggests that the Western blot is an accurate test to survey INH- and RIF-resistant TB antigens of M. tuberculosis infection. These findings indicate that MDR-TB diagnosis (based on Ag detection) could be useful in the identification of disease stages that precede symptomatic and microbiologically positive TB, such as subclinical and incipient TB.

  9. 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...

  10. Interpretation criteria for standardized Western blots for three European species of Borrelia burgdorferi sensu lato.

    PubMed

    Hauser, U; Lehnert, G; Lobentanzer, R; Wilske, B

    1997-06-01

    Western blots (WBs; immunoblots) are a widely used tool for the serodiagnosis of Lyme borreliosis, but so far, no defined criteria for performance, analysis, and interpretation have been established in Europe. For the current study WBs were produced with strains PKa2 (Borrelia burgdorferi sensu stricto), PKo (Borrelia afzelii), and PBi (Borrelia garinii). To improve resolution we used gels of 17 cm in length. In a first step, 13 immunodominant proteins were identified with monoclonal antibodies. Then, the apparent molecular masses of all visually distinguishable bands were determined densitometrically. Approximately 40 bands of between 14 and 100 kDa were differentiated for each strain. From a study with 330 serum samples (from 189 patients with Lyme borreliosis and 141 controls), all observed bands were documented. To establish criteria for a positive WB result, the discriminating ability of a series of band combinations (interpretation rules) were evaluated separately for each strain (for immunoglobulin G [IgG] WB, > 40 combinations; for IgM WB, > 15 combinations). The following interpretation criteria resulting in specificities of greater than 96% were recommended: for IgG WB, at least one band of p83/100, p58, p56, OspC, p21, and p17a for PKa2; at least two bands of p83/100, p58, p43, p39, p30, OspC, p21, p17, and p14 for PKo; and at least one band of p83/100, p39, OspC, p21, and p17b for PBi; for IgM WB, at least one band of p39, OspC, and p17a or a strong p41 band for PKa2; at least one band of p39, OspC, and p17 or a strong p41 band for PKo; and at least one band of p39 and OspC or a strong p41 band for PBi. The overall sensitivity was the highest for PKo WB, followed by PBi and PKa2 WB, in decreasing order. Standardization of WB assays is necessary for comparison of results from different laboratories.

  11. Analysis of Gene and Protein Expression in Atherosclerotic Mouse Aorta by Western Blot and Quantitative Real-Time PCR.

    PubMed

    Rivera-Torres, José

    2015-01-01

    Atherosclerosis involves changes in gene and protein expression patterns in affected arteries. Quantification of these alterations is essential for understanding the molecular mechanisms underlying this pathology. Western blot and real-time PCR-used to quantify protein and messenger RNA levels, respectively-are invaluable molecular biology tools, particularly when material is limited. The availability of many genetically modified mouse models of atherosclerosis makes the mouse aorta an ideal tissue in which to carry out these expression pattern analyses. In this chapter, protocols are presented for mRNA and protein extraction from mouse aorta and for the accurate quantification of mRNA expression by RT-PCR and of proteins by western blot.

  12. Western Blot Assay Using Recombinant p26 Antigen for Detection of Equine Infectious Anemia Virus-Specific Antibodies▿

    PubMed Central

    Alvarez, I.; Gutierrez, G.; Ostlund, E.; Barrandeguy, M.; Trono, K.

    2007-01-01

    We analyzed the performance of a single-band Western blot (WB) test using recombinant p26 (rp26) capsid protein of equine infectious anemia virus. According to the results obtained, the rp26 WB test is a reliable confirmatory diagnostic tool to be used as a complementary test after an enzyme-linked immunosorbent assay or agar gel immunodiffusion test yielding doubtful results. PMID:17959820

  13. 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

  14. Western blot analysis of a limited number of cells: a valuable adjunct to proteome analysis of paraffin wax-embedded, alcohol-fixed tissue after laser capture microdissection.

    PubMed

    Martinet, Wim; Abbeloos, Vanessa; Van Acker, Nathalie; De Meyer, Guido R Y; Herman, Arnold G; Kockx, Mark M

    2004-03-01

    In recent years, laser capture microdissection (LCM) has been used successfully to obtain distinct populations of cells for subsequent molecular analysis. Because of the limited sample availability and the absence of in vitro amplification steps for proteins, the use of LCM for proteome analysis largely depends on highly sensitive protein detection methods. In this study, a western blot protocol was developed and validated for the detection of beta-actin and the moderately expressed cell death protein caspase-3 in small numbers of cells. Initially, cultured human U937 monocytes and whole sections of paraffin wax-embedded, alcohol-fixed human tonsils were used to optimize protein electrophoresis and western blotting conditions. High-performance NuPAGE Bis-Tris gels in combination with high-quality transfer membranes, optimized antibody concentrations, and a sensitive chemiluminescent substrate provided a strong signal for beta-actin with approximately 500 U937 cells. In the same way, procaspase-3 could be identified with approximately 1000 cells. Similar results were obtained with germinal centre cells that were procured from paraffin wax-embedded, alcohol-fixed human tonsils by LCM. Treatment of U937 cells with etoposide rapidly induced cell death and allowed the detection of active caspase-3 with approximately 2500 cells (0.8 pg of protein). The findings of this study suggest that western blotting is a valuable adjunct to proteome analysis of LCM procured cells.

  15. Glycosylated VCAM-1 isoforms revealed in 2D western blots of HUVECs treated with tumoral soluble factors of breast cancer cells

    PubMed Central

    2009-01-01

    Background Several common aspects of endothelial phenotype, such as the expression of cell adhesion molecules, are shared between metastasis and inflammation. Here, we analyzed VCAM-1 variants as biological markers of these two types of endothelial cell activation. With the combination of 2-DE and western blot techniques and the aid of tunicamycin, we analyzed N-glycosylation variants of VCAM-1 in primary human endothelial cells stimulated with either TNF or tumoral soluble factors (TSF's) derived from the human breast cancer cell line ZR75.30. Results Treatments induced a pro-adhesive endothelial phenotype. 2D western blots analysis of cells subjected to both treatments revealed the expression of the two known VCAM-1 isoforms and of previously unknown isoforms. In particular TSFZR75.30 induced an isoform with a relative molecular mass (Mr) and isoelectric point (pI) of 75-77 kDa and 5.0, respectively. Conclusion The unknown isoforms of VCAM-1 that were found to be overexpressed after treatment with TSF's compared with TNF, could serve as biomarkers to discriminate between inflammation and metastasis. 2D western blots revealed three new VCAM-1 isoforms expressed in primary human endothelial cells in response to TSF stimulation. Each of these isoforms varies in Mr and pI and could be the result of differential glycosylation states. PMID:19930605

  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. Hydrogel Pore-Size Modulation for Enhanced Single-Cell Western Blotting.

    PubMed

    Duncombe, Todd A; Kang, Chi-Chih; Maity, Santanu; Ward, Toby M; Pegram, Mark D; Murthy, Niren; Herr, Amy E

    2016-01-13

    Pore-gradient microgel arrays enable thousands of parallel high-resolution single-cell protein electrophoresis separations for targets accross a wide molecular mass (25-289 kDa), yet within 1 mm separation distances. Dual crosslinked hydrogels facilitate gel-pore expansion after electrophoresis for efficient and uniform immunoprobing. The photopatterned, light-activated, and acid-expandable hydrogel underpins single-cell protein analysis, here for oncoprotein-related signaling in human breast biopsy.

  18. 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

  19. Detection of anti-HIV-1 IgG antibodies in whole saliva by GACELISA and Western blot assays.

    PubMed

    Matee, M I; Lyamuya, E F; Simon, E; Mbena, E C; Kagoma, C; Samaranayake, L P; Scheutz, F

    1996-05-01

    The present study, based on 158 HIV seropositives and 167 HIV seronegatives, demonstrates that saliva collected with the Omni-SAL device and tested with GACELISA (an IgG antibody capture ELISA) is an effective non-invasive alternative to serum for anti-HIV IgG antibody screening. The study also shows that a conventional serum Western blot kit can be used, with slight modifications, for confirmatory testing of saliva specimens. Collecting saliva with the Omni-SAL device had a very good acceptance rate among Tanzanian subjects, and although this diagnostic method is not yet known by the general public, 65% of the study participants preferred to give saliva instead of blood for HIV testing.

  20. An analysis of beta-lactam-derived antigens on spleen cell and serum proteins by ELISA and Western blotting.

    PubMed

    Warbrick, E V; Thomas, A L; Stejskal, V; Coleman, J W

    1995-11-01

    Penicillins and related beta-lactam antibiotics are known to conjugate to proteins to generate potentially antigenic (haptenic) determinants. In the present study, we used a rabbit polyclonal antibody raised against benzylpenicillin (BP) to investigate the capacity of six penicillins and one cephalosporin to generate haptenic groups in vitro on cultured mouse spleen cells and on serum proteins in the culture medium. All of the drugs tested, namely, BP, amoxicillin (AMX), ampicillin (AMP), cephalothin (CEP), cloxacillin (CLX), flucloxacillin (FLX), and phenoxymethylpenicillin (PMP) generated antigens in a concentration-dependent manner on cell and serum proteins, which could be detected by ELISA, although antigens generated by BP, CEP, FLX, or PMP in either cell- or serum-conjugated form were more readily detected than those generated by AMX, AMP, or CLX. Western blot analysis revealed that BP-derived antigens were generated relatively slowly on cell proteins (maximum binding was not yet reached after 8 h), compared to serum proteins (maximum binding within 1 h). BP, CEP, and PMP all generated similar distinctive patterns of immunostaining of electrophoresed cell or serum proteins which did not reflect the relative abundance of different proteins as revealed by Coomassie brilliant blue staining. FLX, CLX, AMP, and AMX did not generate antigens that could be detected on Western blots. In conclusion, we have shown that various beta-lactam antibiotics generate antigens on cell and serum proteins that can be detected and characterized immunochemically with polyclonal antiserum. Further application of these methods may offer potential for further identification of immunologically relevant cellular and serum antigens generated by these drugs.

  1. 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

  2. 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.

  3. 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

  4. High resolution autoantibody detection by optimized protein G-horseradish peroxidase immunostaining in a western blotting assay.

    PubMed

    Hu, G R; Harrop, P; Warlow, R S; Gacis, M L; Walls, R S

    1997-03-28

    We report a procedure for optimisation of Western blotting using protein G-horseradish peroxidase (protein G-HRP) which avoids the false positive reactions often caused by second antibodies and increases the detection of autoantibodies by protein G conjugate. A number of modifications were investigated. Higher concentrations of serum and protein G-HRP at 1:5 to 1:10 and 1:100, respectively, increased the detection to the same order as that obtained with second antibody systems and gold staining with silver enhancement. The role of various detergents in the procedure was established. 3-[(3-Cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) in incubation with protein G-HRP increased the binding between protein G and immunoglobulin G. Addition of Tween-20 for blocking produced little background so that protein blockers could be avoided. Prolonged incubation with serum increased markedly the sensitivity of the procedure when compared with the recommended 2 h incubation period. Polyvinylidene difluoride membrane provided better transfer effect, lower background and higher mechanical strength than nitrocellulose membrane. The utilization of only one antibody-specific ligand increased the simplicity, reliability, economy, efficiency and specificity of the method. These modifications make this method significantly better for detection and screening for autoantibodies.

  5. 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

  6. Immunofluorescence, enzyme-linked immunosorbent assay, particle agglutination and western blot for the detection of antibody to human immunodeficiency virus type 1.

    PubMed

    Auwanit, W; Ayuthaya, P I; Balachandra, K; Jayavasu, C; Phanthumachinda, B; Ikuta, K; Yamanishi, K; Kanai, K

    1990-03-01

    Immunofluorescence assay (IFA) has been applied for detection of antibody to human immunodeficiency virus type 1 (HIV-1). To compare the IFA with an enzyme-linked immunosorbent assay (ELISA) and particle agglutination (PA), we examined the antibody response to HIV-1 in 475 sera from AIDS, PGL and ARC patients as well as several risk groups and healthy persons by three methods. The positive results by any methods were confirmed by western blot (WB). The results by all methods were well correlated on the sera from 45 asymptomatic male homosexuals and 70 female prostitutes. There were some false positive results by ELISA in the sera from prisoners and healthy persons. Four sera from drug abusers were positive only by PA and IFA and were negative by ELISA. All were WB-inconclusive. Particle agglutination and IFA results were compared with western blot analysis on 208 ELISA-positive sera. All IFA-strongly positive sera (84%) were positive by western blot. The sera with weakly positive, negative and inconclusive results by IFA (16%) were possibly any of positive, inconclusive or negative by western blot. By PA, 200 of 208 (97%) sera were PA-positive and 1% of these sera were WB-inconclusive while the PA-negative sera were either negative or inconclusive by western blot. These results suggested that PA is a simple and sensitive method for screening of HIV-1 antibody while IFA could be a primary confirmatory test and western blot would then be used for confirming any IFA-negative or inconclusive results.

  7. 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

  8. Outbreak of larval Echinococcus multilocularis infection in Japanese monkey (Macaca fuscata) in a zoo, Hokkaido: western blotting patterns in the infected monkeys.

    PubMed

    Sato, Chiaki; Kawase, Shiro; Yano, Shoki; Nagano, Hideki; Fujimoto, Satoshi; Kobayashi, Nobuyuki; Miyahara, Kazuro; Yamada, Kazutaka; Sato, Motoyoshi; Kobayashi, Yoshiyasu

    2005-01-01

    A high prevalence of larval Echinococcus multilocularis (Em) infection was found in zoo primates in Hokkaido, Japan. In October 1997, a Japanese monkey (Macaca fuscata) died and histopathologically diagnosed as alveolar hydatidosis. Serum samples were collected from the remaining Japanese monkeys and examined for antibodies against Em by enzyme-linked immunosorbent assay and western blotting. Serological tests showed 12 more animals of the remaining 57 monkeys were possibly infected. Ultrasonography revealed that nine of these 12 animals had a cystic lesion in the liver. The band patterns of western blotting in the monkeys were very similar to those in human.

  9. Relative performance of Organon kit in comparison to Du Pont for confirmatory serological testing of HIV infection by western blot test in sera from blood donors.

    PubMed

    Aggarwal, R K; Chatterjee, R; Chattopadhya, D; Kumari, S

    1992-06-01

    A total of 32 specimens with different categories of reactivity by Du Pont Western Blot kit comprising of specimens showing full spectrum of HIV-I antigen specific bands, 19 specimens showing total absence of bands and four specimens showing non-specific bands (without any interpretative importance) were subjected to Western Blot testing by Organon test. Of the nine specimens showing full spectrum of bands by Du Pont the correlation with Organon kit was 100 per cent based on WHO criteria. Four specimens with non-specific indeterminate band pattern by Du Pont failed to show any band in Organon kit, indicating that latter to be more specific.

  10. 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)

  11. 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

  12. Immunohistochemical and Western blot analyses of collar enamel in the jaw teeth of gars, Lepisosteus oculatus, an actinopterygian fish.

    PubMed

    Sasagawa, Ichiro; Ishiyama, Mikio; Yokosuka, Hiroyuki; Mikami, Masato; Shimokawa, Hitoyata; Uchida, Takashi

    2014-06-01

    Although most fish have no enamel layer in their teeth, those belonging to Lepisosteus (gars), an extant actinopterygian fish genus, do and so can be used to study amelogenesis. In order to examine the collar enamel matrix in gar teeth, we subjected gar teeth to light and electron microscopic immunohistochemical examinations using an antibody against bovine amelogenin (27 kDa) and antiserum against porcine amelogenin (25 kDa), as well as region-specific antibodies and antiserum against the C-terminus and middle region, and N-terminus of porcine amelogenin, respectively. The enamel matrix exhibited intense immunoreactivity to the anti-bovine amelogenin antibody and the anti-porcine amelogenin antiserum in addition to the C-terminal and middle region-specific antibodies, but not to the N-terminal-specific antiserum. These results suggest that the collar enamel matrix of gar teeth contains amelogenin-like proteins and that these proteins possess domains that closely resemble the C-terminal and middle regions of porcine amelogenin. Western blot analyses of the tooth germs of Lepisosteus were also performed. As a result, protein bands with molecular weights of 78 kDa and 65 kDa were clearly stained by the anti-bovine amelogenin antibody as well as the antiserum against porcine amelogenin and the middle-region-specific antibody. It is likely that the amelogenin-like proteins present in Lepisosteus do not correspond to the amelogenins found in mammals, although they do possess domains that are shared with mammalian amelogenins.

  13. Deep proteomic profiling of vasopressin-sensitive collecting duct cells. I. Virtual Western blots and molecular weight distributions.

    PubMed

    Yang, Chin-Rang; Tongyoo, Pumipat; Emamian, Milad; Sandoval, Pablo C; Raghuram, Viswanathan; Knepper, Mark A

    2015-12-15

    The mouse mpkCCD cell line is a continuous cultured epithelial cell line with characteristics of renal collecting duct principal cells. This line is widely used to study epithelial transport and its regulation. To provide a data resource useful for experimental design and interpretation in studies using mpkCCD cells, we have carried out "deep" proteomic profiling of these cells using three levels of fractionation (differential centrifugation, SDS-PAGE, and HPLC) followed by tandem mass spectrometry to identify and quantify proteins. The analysis of all resulting samples generated 34.6 gigabytes of spectral data. As a result, we identified 6,766 proteins in mpkCCD cells at a high level of stringency. These proteins are expressed over eight orders of magnitude of protein abundance. The data are provided to users as a public data base (https://helixweb.nih.gov/ESBL/Database/mpkFractions/). The mass spectrometry data were mapped back to their gel slices to generate "virtual Western blots" for each protein. For most of the 6,766 proteins, the apparent molecular weight from SDS-PAGE agreed closely with the calculated molecular weight. However, a substantial fraction (>15%) of proteins was found to run aberrantly, with much higher or much lower mobilities than predicted. These proteins were analyzed to identify mechanisms responsible for altered mobility on SDS-PAGE, including high or low isoelectric point, high or low hydrophobicity, physiological cleavage, residence in the lysosome, posttranslational modifications, and expression of alternative isoforms due to alternative exon usage. Additionally, this analysis identified a previously unrecognized isoform of aquaporin-2 with apparent molecular mass <20 kDa.

  14. 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

  15. A mass screening survey of cystic echinococcosis by ultrasonography, Western blotting, and ELISA among university students in Manisa, Turkey.

    PubMed

    Kilimcioğlu, Ali Ahmet; Girginkardeşler, Nogay; Korkmaz, Metin; Özkol, Mine; Düzgün, Fatih; Östan, Ipek; Pabuşcu, Yüksel; Dinç, Gönül; Ok, Ulgen Zeki

    2013-12-01

    Cystic echinococcosis (CE) is one of the most important zoonotic diseases in a wide geographic area, including Turkey. In the present project, a total of 4275 students from Celal Bayar University, Manisa, Turkey, were screened by ultrasonography (US) and specific antibodies for CE were examined by Western blotting (WB) and ELISA in finger prick blood samples of 2034 of 4275 volunteered students. We aimed to report the apparent prevalence of CE based on different diagnostic procedures and to compare WB and ELISA with US in diagnosis of CE in a mass screening setting. Six new cases were diagnosed as CE by US during the survey. In addition to these cases, three students were also detected to have been previously operated and pathologically confirmed for hepatic CE. US revealed parenchymal changes in these cases in concordance with their operation history; so, the prevalence of CE by US was calculated as 0.21% (9/4275) (95%CI, 0.11-0.39%) among university students in Manisa. Bands were detected at 8, 28, 32, 38, 42, 47, 70 and 90kDa by WB and the cases were considered to be positive for CE when at least three of the bands were seen together. Apparent prevalence of CE by ELISA and WB were found to be 2.11% (43/2034) (95%CI, 1.57-2.83%) and 0.25% (5/2034) (95%CI, 0.10-0.57%), respectively. Of the six US positive cases, WB was positive in only one case with two cysts in the liver. All of four cases with liver involvement were positive by ELISA. The high prevalence of CE among university students in Manisa indicated that CE is a major health problem in this area of Turkey. Our results supported that WB is rather difficult and not feasible as a mass screening test and may not be effective for confirmation especially in asymptomatic cases. As a result, we recommend US to be used initially in mass screening surveys for CE followed by confirmation by ELISA for suspected cases. Further examination primarily by chest X-ray followed by computed tomography and/or magnetic

  16. Validity of interpretation criteria for standardized Western blots (immunoblots) for serodiagnosis of Lyme borreliosis based on sera collected throughout Europe.

    PubMed

    Hauser, U; Lehnert, G; Wilske, B

    1999-07-01

    Western blotting (WB; immunoblotting) is a widely used tool for the serodiagnosis of Lyme borreliosis (LB), but so far, no generally accepted criteria for performance and interpretation have been established in Europe. The current study was preceeded by a detailed analysis of WB with whole-cell lysates of three species of Borrelia burgdorferi sensu lato (U. Hauser, G. Lehnert, R. Lobentanzer, and B. Wilske, J. Clin. Microbiol. 35:1433-1444, 1997). In that study, interpretation criteria for a positive WB result were developed with the data for 330 serum samples (from patients with LB in different stages [n = 189] and from a control group [n = 141]) originating mostly from southern Germany. In the present work, the interpretation criteria for strains PKo (Borrelia afzelii) and PBi (Borrelia garinii) developed in the previous study were reevaluated with 224 serum samples (from patients with LB in different stages [n = 97] and from a control group [n = 127]) originating from throughout Europe that were provided by the European Union Concerted Action on Lyme Borreliosis (EUCALB). De novo criteria were developed on the basis of the reactivities of the EUCALB sera and were evaluated with the data for the samples from southern Germany. Comparison of all results led to the following recommendations: For WB for immunoglobulin G (IgG), at least two bands among p83/100, p58, p43, p39, p30, OspC, p21, p17, and p14 for PKo and at least one band among p83/100, p39, p30, OspC, p21, and p17b for PBi; for WB for IgM, at least one band among p39, OspC, and p17 or a strong p41 band for PKo and at least one band among p39 and OspC or a strong p41 band for PBi. WB with PKo was the most sensitive, and this strain is recommended for use in WB for the serodiagnosis of LB throughout Europe.

  17. Southern blotting.

    PubMed

    Brown, T

    2001-05-01

    Southern blotting is the transfer of DNA fragments from an electrophoresis gel to a membrane support, 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 unit describes Southern blotting via upward capillary transfer of DNA from an agarose gel onto a nylon or nitrocellulose membrane, and subsequent immobilization 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. 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 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 the second alternate protocol is therefore more rapid and can result in more complete transfer. PMID:18432697

  18. 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

  19. A modified ELISA and western blot accurately determine anti-human immunodeficiency virus type 1 antibodies in oral fluids obtained with a special collecting device.

    PubMed

    Emmons, W W; Paparello, S F; Decker, C F; Sheffield, J M; Lowe-Bey, F H

    1995-06-01

    Serum and saliva from 195 known human immunodeficiency virus (HIV)-seropositive patients and 198 military personnel undergoing annual HIV serologic testing were evaluated in a prospective, blinded fashion for anti-HIV-1 antibodies. Oral specimens, collected with a device designed to concentrate oral mucosal transudate from whole saliva, were tested by a modified ELISA and by Western blot. Serum was tested in a standard manner. All 195 HIV-1-seropositive subjects had detectable anti-HIV-1 antibodies in their saliva by ELISA; 190 saliva samples were positive by Western blot and 5 were indeterminate. None of the 198 military personnel were positive by ELISA of serum or oral fluid. The sensitivity, specificity, and positive and negative predictive values for ELISA of saliva were each 100%. The serologic testing of oral mucosal transudate appears to be a simple, safe, sensitive, and specific method for detecting anti-HIV-1 antibodies.

  20. 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

  1. 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

  2. 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

  3. The seroprevalence of Taenia solium cysticercosis among epileptic patients in León, Nicaragua, as evaluated by ELISA and western blotting.

    PubMed

    Bucardo, F; Meza-Lucas, A; Espinoza, F; García-Jerónimo, R C; García-Rodea, R; Correa, D

    2005-01-01

    The Taenia solium taeniasis/cysticercosis complex is an important public-health problem in several countries, where many epileptic seizures appear to be associated with neurocysticercosis. As few data on this problem in Nicaragua exist, the seroprevalence of antibodies reacting with antigens from T. solium cysticerci was investigated among 88 Nicaraguan epileptics (45 males and 43 females, aged 6-53 years). In questionnaire-based interviews, each adult subject and a caregiver of each child investigated were asked about potential risk factors for taeniasis/cysticercosis. When a serum sample from each subject was then checked for anti-cysticercus antibodies, 8.0% of the subjects were found seropositive by ELISA and 14.8% by western blotting. Five samples (all from individuals who had been epileptic for > 5 years) were positive in both tests. When the level of association between each potential risk factor and seropositivity (in ELISA or by blotting) was evaluated, the only statistically significant association detected was that between a positive ELISA and the subject living in a household where pigs were raised (odds ratio = 5.18; 95% confidence interval = 0.8-41.6; P = 0.05). The bands most frequently recognized in the western blots (of 50, 42-39, 24 and 14 kDa) were those previously reported. The results indicate that, in the city of Léon, cysticercosis may be endemic and the cause of a significant proportion of the epilepsy recorded.

  4. Evaluation of viral-lysate enzyme-linked immunosorbent assay kits for detecting human immunodeficiency virus (type 1) infections using human sera standardized by quantitative western blotting.

    PubMed

    Hardy, C T; Damrow, T A; Villareal, D B; Kenny, G E

    1992-06-01

    The first generation of proprietary reagents for detecting antibodies to the Human Immunodeficiency Virus Type 1 (HIV-1) by enzyme-linked immunosorbent assay (ELISA) used as antigen partially purified virus from cell culture lysates. These tests, which are still in use, may vary in their antibody measurement capabilities if different proportions of the viral polypeptides are present in the viral lysate mixtures. We determined the quantities of antibodies in the serum of persons infected with HIV-1 by dilution analysis using 3 ELISA kits: Abbott [A], Du Pont [D], Genetic Systems [G]. The proportionate antibody titres of each serum to p24gag and gp160env/120env were established by quantitative Western blotting. Serum antibody titres were high, frequently over 1:10,000, a result observed both by ELISA and Western blot. For Kit D, sera with high proportions of antibody to p24gag produced antibody titration curves with steep slopes whereas shallower slopes were found in sera with high proportions of antibody to gp160env. In contrast, Kit A gave steeper slopes with sera enriched for gp160env antibodies. Kit G gave results with slopes intermediate between Kits A and D. Serum antibody titres differed between kits depending upon the proportion and concentration of antibodies in a given serum to gp160env and p24gag. The findings that both the concentration and proportion of antibodies to specific viral polypeptides in human sera markedly affect the signal intensity produced by proprietary ELISAs suggest the need for several control sera which reflect the diversity of human serum responses. Standardization of human reference sera by quantitative Western blotting will assist in evaluation and quality control of ELISA tests.

  5. Study of a viral-dual infection in rainbow trout (Oncorhynchus mykiss) by seroneutralization, western blot and polymerase chain reaction assays.

    PubMed

    Rodríguez, S; Vilas, M P; Alonso, M; Pérez, S I

    1995-12-01

    Viral-dual infections in fish are of interest to aquaculture practices but they are rarely described and studied. Several methods were applied in this work to demonstrate a case of coinfection in a reared rainbow trout (Oncorhynchus mykiss) population. Inoculation in cell cultures and cross-neutralization tests were the standard procedures that made it possible to isolate and identify a birnavirus, the infectious pancreatic necrosis virus (IPNV), and suspect of a second virus. Western blotting with both polyclonal and monoclonal antibodies, and reverse transcriptional-polymerase chain reaction (RT-PCR) demonstrate coexistence of both, IPNV and a rhabdovirus.

  6. Western blot analysis of Amblyomma americanum-derived stage-specific and shared antigens using serum from guinea pigs expressing resistance.

    PubMed

    Brown, S J

    1988-04-01

    Serum from guinea pigs expressing resistance to larval, nymphal and adult Amblyomma americanum ticks was used in Western blot analyses to identify potential antigens from egg, larval and nymphal, and female salivary gland extract preparations. The results demonstrate multiple antigens unique to each life stage, as well as several shared proteins between the three life stages. However, it appears as if two particular proteins of 25 and 38 kDa may be more important than others, based upon their prevalence and intensity of recognition in this assay relative to other polypeptides. PMID:2455376

  7. Rapid Preparation of a Plasma Membrane Fraction: Western Blot Detection of Translocated Glucose Transporter 4 from Plasma Membrane of Muscle and Adipose Cells and Tissues.

    PubMed

    Yamamoto, Norio; Yamashita, Yoko; Yoshioka, Yasukiyo; Nishiumi, Shin; Ashida, Hitoshi

    2016-01-01

    Membrane proteins account for 70% to 80% of all pharmaceutical targets, indicating their clinical relevance and underscoring the importance of identifying differentially expressed membrane proteins that reflect distinct disease properties. The translocation of proteins from the bulk of the cytosol to the plasma membrane is a critical step in the transfer of information from membrane-embedded receptors or transporters to the cell interior. To understand how membrane proteins work, it is important to separate the membrane fraction of cells. This unit provides a protocol for rapidly obtaining plasma membrane fractions for western blot analysis. © 2016 by John Wiley & Sons, Inc. PMID:27479506

  8. Localization of α-synuclein in teleost central nervous system: immunohistochemical and Western blot evidence by 3D5 monoclonal antibody in the common carp, Cyprinus carpio.

    PubMed

    Vaccaro, Rosa; Toni, Mattia; Casini, Arianna; Vivacqua, Giorgio; Yu, Shun; D'este, Loredana; Cioni, Carla

    2015-05-01

    Alpha synuclein (α-syn) is a 140 amino acid vertebrate-specific protein, highly expressed in the human nervous system and abnormally accumulated in Parkinson's disease and other neurodegenerative disorders, known as synucleinopathies. The common occurrence of α-syn aggregates suggested a role for α-syn in these disorders, although its biological activity remains poorly understood. Given the high degree of sequence similarity between vertebrate α-syns, we investigated this proteins in the central nervous system (CNS) of the common carp, Cyprinus carpio, with the aim of comparing its anatomical and cellular distribution with that of mammalian α-syn. The distribution of α-syn was analyzed by semiquantitative western blot, immunohistochemistry, and immunofluorescence by a novel monoclonal antibody (3D5) against a fully conserved epitope between carp and human α-syn. The distribution of 3D5 immunoreactivity was also compared with that of choline acetyltransferase (ChAT), tyrosine hydroxylase (TH), and serotonin (5HT) by double immunolabelings. The results showed that a α-syn-like protein of about 17 kDa is expressed to different levels in several brain regions and in the spinal cord. Immunoreactive materials were localized in neuronal perikarya and varicose fibers but not in the nucleus. The present findings indicate that α-syn-like proteins may be expressed in a few subpopulations of catecholaminergic and serotoninergic neurons in the carp brain. However, evidence of cellular colocalization 3D5/TH or 3D5/5HT was rare. Differently, the same proteins appear to be coexpressed with ChAT by cholinergic neurons in several motor and reticular nuclei. These results sustain the functional conservation of the α-syn expression in cholinergic systems and suggest that α-syn modulates similar molecular pathways in phylogenetically distant vertebrates.

  9. Advanced laboratory techniques for diagnosing Toxoplasma gondii encephalitis in AIDS patients: significance of intrathecal production and comparison with PCR and ECL-western blotting.

    PubMed

    Contini, C; Fainardi, E; Cultrera, R; Canipari, R; Peyron, F; Delia, S; Paolino, E; Granieri, E

    1998-12-01

    The polymerase chain reaction (PCR) for detection of cerebral spinal fluid (CSF) Toxoplasma gondii DNA was combined with the study of intrathecal antibody synthesis by antibody specific index calculation (ASI) and the detection of specific oligoclonal IgG bands (OCB) by affinity mediated immunoblotting (AMI) in 11 AIDS patients with T. gondii encephalitis (TE) and in 20 control patients with or without neurological disorders. Enhanced chemiluminescence (ECL) western-blot technique was employed to evaluate the antigenic specificity of CSF-IgG towards individual T. gondii antigens. PCR was positive in all TE patients which displayed brain-derived or blood-derived specific OCB, even when comparative ASI failed. Four TE patients had a unique anti-T. gondii OCB restricted to the CSF and a strong antibody response toward the 29 kDa band by ECL western blot. This response could be an important marker to discriminate TE from other opportunistic central nervous system (CNS) infections in the course of AIDS. PMID:9916877

  10. Current state of HIV/AIDS in Taiwan and reappraisal of early diagnostic value of serial and comparative analyses of western blot patterns of HIV infection.

    PubMed

    Chen, Mao-Yuan; Chuang, Che-Yen

    2002-02-01

    Since Taiwan is not a member of the World Health Organization (WHO), the problems of human immunodeficiency virus (HIV) and acquired immunodeficiency syndrome (AIDS) in Taiwan are not well known to the world. In this report we summarize the trend and current status of HIV/AIDS in Taiwan and also analyze Western blot (WB) patterns. The application of serial and comparative WB analyses is important in establishing an early diagnosis of neonatal HIV infection. These analyses will be useful as well in evaluating the clinical status of adults infected with HIV in developing countries. When there are "indeterminate" WBs in adults with recent HIV infection, it is imperative to perform a WB weekly to attain the earliest possible diagnosis and treatment. Usually anti-gag antibodies, most frequently the anti-p24, are followed by anti-env and anti-pol antibodies. When the set point is attained, WB studies may also be repeated if the clinical status changes or if the patients are receiving highly active antiretroviral therapy (HAART). In the advanced stages of HIV infection, usually the anti-gag antibodies will fade first and then disappear, this occurs next in the anti-pol and in the anti-env antibodies. During HAART, viral replication is usually controlled and there will thus be a partial restoration of the immunological function that will induce changes in the antigen-antibody ratio and, in the end, this will result in changes in the WB patterns. In the mid 1980s all the hepatitis B immunoglobulin (HBIG) used in Taiwan was anti-HIV positive. However, follow-up ELISA and WB studies of the HBIG injected newborns proved that HBIG was anti-HIV positive but that there was no replicable HIV in HBIG. Monthly WB tests of newborns and anti-HIV positive mothers were used to differentiate HIV infection from the passive placental transference by comparing their WB patterns. Comparative WB analyses was also done between blood donors and recipients, and also between husbands and wives

  11. 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

  12. 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

  13. 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.

  14. 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

  15. 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

  16. 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

  17. 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

  18. Interrogation of multidrug resistance (MDR1) P-glycoprotein (ABCB1) expression in human pancreatic carcinoma cells: correlation of 99mTc-Sestamibi uptake with western blot analysis.

    PubMed

    Harpstrite, Scott E; Gu, Hannah; Natarajan, Radhika; Sharma, Vijay

    2014-10-01

    Histopathological studies indicate that ∼63% of pancreatic tumors express multidrug resistance (MDR1) P-glycoprotein (Pgp) and its polymorphic variants. However, Pgp expression detected at the mRNA or protein level does not always correlate with functional transport activity. Because Pgp transport activity is affected by specific mutations and the phosphorylation state of the protein, altered or less active forms of Pgp may also be detected by PCR or immunohistochemistry, which do not accurately reflect the status of tumor cell resistance. To interrogate the status of the functional expression of MDR1 Pgp in MiaPaCa-2 and PANC-1 cells, cellular transport studies using Tc-Sestamibi were performed and correlated with western blot analysis. Biochemical transport assays in human pancreatic carcinoma MiaPaCa-2 and PANC-1 cells, human epidermal carcinoma drug-sensitive KB-3-1 cells, and human breast carcinoma MCF-7 cells (negative controls), and human epidermal carcinoma drug-resistant KB-8-5 cells, human breast carcinoma stably transfected with Pgp MCF-7/MDR1Pgp cells, and liver carcinoma HepG2 cells (positive controls) were performed. Protein levels were determined using a monoclonal antibody C219. Tc-Sestamibi demonstrates accumulation in human pancreatic carcinoma MiaPaCa-2 and PANC-1 cells. Uptake profiles are not affected by treatment with LY335979, a Pgp inhibitor, and correlate with western blot analysis. These cellular transport studies indicate an absence of Pgp at a functional level in MiaPaCa-2 and PANC-1 cells. Because major pancreatic tumors originate from the pancreatic duct and Tc-Sestamibi undergoes a dominant hepatobiliary mode of excretion, it would not be a sensitive probe for imaging pancreatic adenocarcinomas. Following interrogation of the functional status of Pgp in other pancreatic carcinoma cells, chemotherapeutic drugs that are also MDR1 substrates could offer alternative therapeutics for treating pancreatic adenocarcinomas.

  19. A comparative study of Toxoplasma gondii seroprevalence in mink using a modified agglutination test, a Western blot, and enzyme-linked immunosorbent assays.

    PubMed

    Gu, Yi; Wang, Zedong; Cai, Yufeng; Li, Xiaoxing; Wei, Feng; Shang, Limin; Li, Jiping; Liu, Quan

    2015-09-01

    Toxoplasma gondii can infect almost all warm-blooded animals, and many serological methods have been developed to detect T. gondii infection in a variety of animal species. In the present study, the seroprevalence of T. gondii infection in farmed mink in northeast China was determined using the modified agglutination test (MAT), a Western blot (WB), and 3 enzyme-linked immunosorbent assays (ELISAs) with protein A/G conjugate, using either of 2 recombinant dense granule antigens, GRA1 and GRA7, or Toxoplasma soluble antigens (TSA). There was no significant difference between the detection results of the GRA1-, GRA7-, and TSA-ELISAs and WB (McNemar chi-square, P > 0.05), but a significant difference was observed between MAT and WB (P < 0.05). A near perfect agreement (97.0%) was found between the GRA7-ELISA and WB (κ = 0.83), and a substantial agreement (92.4-93.1%) was observed in the TSA- and GRA1-ELISAs (κ = 0.68-0.73). The GRA7-ELISA showed the highest sensitivity and specificity, and the lowest false-positive and negative rates, while the MAT gave both a low sensitivity and frequent false positives in comparison to the WB. Receiver operating characteristic analysis revealed the largest area under curve of 0.85 (95% confidence interval: 0.74-0.96), and the highest relative sensitivity (72.7%) and specificity (99.0%) for a cutoff value of 0.19 in the GRA7-ELISA. These results indicate that the GRA7-ELISA is suitable for detection of T. gondii infection in mink and that MAT should be used with caution.

  20. Evaluation of the Bio-Rad Geenius HIV 1/2 Confirmation Assay as an Alternative to Western Blot in the Korean Population: A Multi-Center Study.

    PubMed

    Moon, Hee-Won; Huh, Hee Jin; Oh, Gwi Young; Lee, Sang Gon; Lee, Anna; Yun, Yeo-Min; Hur, Mina

    2015-01-01

    Recently updated recommendations for diagnosis of HIV infection suggest a new diagnostic algorithm including HIV-1/HIV-2 antibody differentiation immunoassay instead of western blot (WB) as a confirmatory testing. We evaluated Bio-Rad Geenius HIV1/2 confirmation assay as a simple and reliable alternative to WB in the Korean population with low HIV prevalence. The Geenius HIV1/2 was performed in a total of 192 serum specimens (140 reactive and 52 nonreactive specimens by ARCHITECT HIV Ag/Ab Combo assay) that were prospectively collected from five institutions. HIV-1 nucleic acid amplification test (NAT) was performed in negative or indeterminate specimens by Geenius HIV1/2 or WB. Among 140 reactive specimens by HIV Ag/Ab assay, 82 (58.6%) were positive for HIV-1 Ab by Geenius HIV1/2. Among 58 negative or indeterminate specimens by Geenius HIV1/2, four specimens (6.9%) were positive by HIV-1 NAT. The sensitivity and specificity of Geenius HIV1/2 were 95.3% and 100.0%, respectively. When we considered only WB, the sensitivity and specificity of Geenius HIV1/2 were 100.0% and 99.1%, respectively. Agreement between Geenius HIV1/2 and WB was excellent (weighted Kappa = 0.89). The Geenius HIV1/2 is simple and time-saving compared with WB. It has an excellent performance and can be a reliable alternative to WB. HIV-1 NAT should be performed in negative or indeterminate specimens by Geenius HIV1/2 to detect acute HIV infection as recommended in new HIV testing algorithms.

  1. Comparison of Enzyme-Linked Immunosorbent Assay, Western Blotting, Microagglutination, Indirect Immunofluorescence Assay, and Flow Cytometry for Serological Diagnosis of Tularemia

    PubMed Central

    Porsch-Özcürümez, Mustafa; Kischel, Nele; Priebe, Heidi; Splettstösser, Wolf; Finke, Ernst-Jürgen; Grunow, Roland

    2004-01-01

    The serodiagnostic efficiencies of five different approaches to detecting antibodies (immunoglobulins G, A, and M) developed in clinically proven infections with Francisella tularensis have been assessed. Fifty serum samples from patients suffering from tularemia during an outbreak in Sweden were compared with samples from 50 healthy blood donors (controls) by using an enzyme-linked immunosorbent assay (ELISA), microagglutination (MA), Western blotting (WB), an indirect immunofluorescence assay (IIFA), and flow cytometry (FC). ELISA, WB, and FC were based on the use of preparations of lipopolysaccharides (LPS) of the live vaccine strain of Francisella tularensis subsp. holarctica (ATCC 29684) as a capture antigen. Whole methanol-fixed bacteria were used for IIFA and MA. Optimized protocols yielded a diagnostic sensitivity and specificity of 100% for WB, MA, and FC, 98% for ELISA, and 93% for IIFA. A total of 6,632 serum samples from individuals between the ages of 18 and 79 years, representatively recruited from all regions of Germany, were screened to estimate and confirm the positive predictive value (PVpos) of the ELISA. Serum samples from 15 (0.226%) individuals tested positive for F. tularensis-specific antibodies by ELISA and confirmatory WB. The resulting prevalence-dependent PVpos of 10.2% and specificity of 98.1% were consistent with our findings for tularemia patients and controls. We conclude that the combined usage of a screening ELISA and a confirmatory WB based on LPS as a common antigen, as well as the MA, is a suitable serodiagnostic tool, while the quality of the IIFA is hampered by subjective variations of the results. FC is a promising new approach that might be improved further in terms of multiplex analyses or high-throughput applications. PMID:15539498

  2. 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

  3. Identification of immunoreactive proteins of Chlamydia trachomatis by Western blot analysis of a two-dimensional electrophoresis map with patient sera.

    PubMed

    Sanchez-Campillo, M; Bini, L; Comanducci, M; Raggiaschi, R; Marzocchi, B; Pallini, V; Ratti, G

    1999-08-01

    Western blots of two-dimensional electrophoretic maps of proteins from Chlamydia trachomatis were probed with sera from 17 seropositive patients with genital inflammatory disease. Immunoblot patterns (comprising 28 to 2 spots, average 14.8) were different for each patient; however, antibodies against a spot-cluster due to the chlamydia-specific antigen outer membrane protein-2 (OMP2) were observed in all sera. The next most frequent group of antibodies (15/17; 88%) recognized the hsp60 GroEL-like protein, described as immunopathogenic in chlamydial infections. Reactivity to the major surface-exposed and variable antigen major outer membrane protein (MOMP) was observed at a relatively lower frequency (13/17; 76%). The hsp70 DnaK-like protein was also frequently recognized (11/17; 64.7%) in this patient group. Besides the above confirmatory findings, the study detected several new immunoreactive proteins, with frequencies ranging from 11/17 to 1/17. Some were characterized also by N-terminal amino acid sequencing and homology searches. Amongst these were a novel outer membrane protein (OmpB) and, interestingly, five conserved bacterial proteins: four (23%) sera reacted with the RNA polymerase alpha-subunit, five (29%) recognized the ribosomal protein S1, eight (47%) the protein elongation factor EF-Tu, seven (41%) a putative stress-induced protease of the HtrA family, and seven sera (41%) the ribosomal protein L7/L12. Homologs of the last two proteins were shown to confer protective immunity in other bacterial infections. The data show that immunological sensitization processes commonly thought to play a role in chlamydial pathogenicity may be sustained not only by the hsp60 GroEl-like protein, but also by other conserved bacterial antigens, some of which may be also considered as potential vaccine candidates. PMID:10493131

  4. 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

  5. Detection of antibody to Toxocara vitulorum perieneteric fluid antigens (Pe) in the colostrum and serum of buffalo calves and cows by Western blotting.

    PubMed

    Ferreira, Fabiano P; Starke-Buzetti, Wilma A

    2005-04-20

    Toxocara vitulorum, a nematode parasite in the small intestine of cattle and water buffaloes, causes high morbidity and mortality of 1-3 months old buffalo calves. This research evaluated the specific perieneteric antigens (Pe) reactivity of anti-T. vitulorum-Pe antibody (Tv-Pe-Ab) in both immune sera and colostrum from buffalo cows immediately post-partum from buffalo cows. The presence of Tv-Pe-Ab in sera of buffalo newborn calves was also examined at 1 day before and after suckling the colostrum as well as in sera from naturally infected calves at the beginning and peak of the maximum infection and then again during the period of rejection and post-rejection of the parasite. Pe antigens were characterized for Tv-Pe-Ab by SDS-PAGE and Western blot (WB). The SDS-PAGE showed that Pe contained nine protein bands (11, 14, 31, 38, 58, 76, 88, 112 and 165 kDa). All Pe bands were recognized by Tv-Pe-Ab in sera and colostrum of buffalo cows. Only the serum antibodies of buffalo calves at 1 day of age after suckling the colostrum and during the beginning of T. vitulorum infection recognized Pe antigen's nine bands. In contrast, serum antibodies from 1-day-old buffalo calves, taken before suckling colostrum, did not react with any protein band. In suckling calves, which reached peak egg output, rejection and post-rejection stages of the infection, serum Tv-Pe-Ab reactivity with lower molecular weight protein bands (11-76 kDa) was lost and only reactivity with the Pe protein bands of higher molecular weight (88, 112 and 165 kDa) remained.

  6. Analysis of the serologic relationship among San Miguel sea lion virus and vesicular exanthema of swine virus isolates. Application of the western blot assay for detection of antibodies in swine sera to these virus types.

    PubMed

    Seal, B S; House, J A; Whetstone, C A; Neill, J D

    1995-04-01

    Caliciviruses are positive-sense single-stranded RNA viruses with a single capsid protein. The serotypes of the marine mammal calicivirus, San Miguel sea lion virus (SMSV), are antigenically related to vesicular exanthema of swine virus (VESV) and are potentially hazardous to swine. Western blot assays using purified SMSV serotypes 1 and 4 were used to further examine the serologic relationship among SMSV and VESV isolates. With the exception of SMSV 8 and SMSV 12, rabbit polyclonal antisera generated against all the available SMSV and VESV isolates reacted positively, as assessed by western blot, with purified capsid protein from SMSV 1 and SMSV 4. Consequently, the SMSV 8 and SMSV 12 virus isolates may not be members of the SMSV/VESV calicivirus group. Using antisera from pigs experimentally inoculated with SMSV and VESV as positive controls, a western blot assay for these virus types was utilized to check for the presence of antibodies to calciviruses in swine sera. Sera from colostrum-deprived gnotobiotic pigs were used as a negative control in all experiments. Examination of sera from domestic and feral swine collected in Iowa, California, and Florida was completed using this technique. The presence of antibodies to these virus types was not detected in any of the porcine sera tested. PMID:7619900

  7. 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,…

  8. Mutation detection by Southern blotting.

    PubMed

    Mellars, Gillian; Gomez, Keith

    2011-01-01

    Following the discovery of the structure of DNA in 1953, it became clear that scientists needed to be able to distinguish different DNA sequences. In 1975, Edward Southern published details of a new method for detecting DNA fragments based upon their specific sequence [corrected]. An indication of the importance of his work is that the technique was eponymously named after him and that subsequent methods based loosely on similar principles were named using a play on his surname (western and northern blot). The simplicity and effectiveness of the technique led to its universal acceptance as a standard method for identification of DNA sequences. In the modern laboratory where turn-around times assume ever greater importance, the process can seem relatively time-consuming. In some cases, this has led to its replacement by more rapid techniques such as long-range PCR. Nevertheless, more than 30 years after its invention, the Southern blot remains a cornerstone of molecular biology. PMID:20938846

  9. Collagen chains detected by western blotting using a /sup 125/I-labeled 45K fragment of fibronectin (45K FN)

    SciTech Connect

    Ristagno, R.; Heimer, R.; Fishman, A.P.; Sampson, P.M.

    1987-05-01

    The objective was to improve the sensitivity and specificity of detection of unlabeled collagen chains in biologic fluids. Chains of Types I,II,III,IV and XI (1..cap alpha..2..cap alpha..3..cap alpha..) collagen were separated by SDS PAGE. Their complete transfer to nitrocellulose was obtained by electrophoresis for 16 h at 150 mA with 10 mM Tris, 117 mM glycine, 100 mM cysteine, 0.1% SDS and 10% methanol. The 45K FN was prepared by chymotryptic digestion of fibronectin adsorbed to gelatin-Sepharose, followed by elution with 1.2 M urea, 1 M Tris-NaCl, pH 8.3 and iodination. When exposed to the nitrocellulose transblot at pH 9.5 and 4/sup 0/C, 45K FN did not react with IgG, fibrinogen, myosin, albumin or carbonic anhydrase. These proteins interfere in the assay under conditions of lower pH and higher temperature. The autoradiographs of the transblots were evaluated by densitometry and reflected results also obtained by dot blotting, that chains of collagen Types I,II,III were detectable at 4 ng and those of collagen Type IV at 12 ng. Generally, ..cap alpha..,BETA, and ..gamma.. chains were detectable. The 45K FN reacted equally with ..cap alpha..1(I) and ..cap alpha..2(I), but for Type XI the 1..cap alpha.. chain had considerably more reactivity than 2..cap alpha.. or 3..cap alpha... As the 45K FN was specific for collagens added to plasma, the authors method appears useful for qualitative and quantitative assays of unlabeled collagens in biologic fluids.

  10. 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.

  11. 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

  12. A brief review of other notable protein blotting methods.

    PubMed

    Kurien, Biji T; Scofield, R Hal

    2009-01-01

    A plethora of methods have been used for transferring proteins from the gel to the membrane. These include centrifuge blotting, electroblotting of proteins to Teflon tape and membranes for N- and C-terminal sequence analysis, multiple tissue blotting, a two-step transfer of low and high molecular weight proteins, blotting of Coomassie Brilliant Blue (CBB)-stained proteins from polyacrylamide gels to transparencies, acid electroblotting onto activated glass, membrane-array method for the detection of human intestinal bacteria in fecal samples, protein microarray using a new black cellulose nitrate support, electrotransfer using square wave alternating voltage for enhanced protein recovery, polyethylene glycol-mediated significant enhancement of the immunoblotting transfer, parallel protein chemical processing before and during western blot and the molecular scanner concept, electronic western blot of matrix-assisted laser desorption/ionization (MALDI) mass spectrometry-identified polypeptides from parallel processed gel-separated proteins, semidry electroblotting of peptides and proteins from acid-urea polyacrylamide gels, transfer of silver-stained proteins from polyacrylamide gels to polyvinylidene difluoride (PVDF) membranes, and the display of K(+) channel proteins on a solid nitrocellulose support for assaying toxin binding. The quantification of proteins bound to PVDF membranes by elution of CBB, clarification of immunoblots on PVDF for transmission densitometry, gold coating of nonconductive membranes before MALDI tandem mass spectrometric analysis to prevent charging effect for analysis of peptides from PVDF membranes, and a simple method for coating native polysaccharides onto nitrocellulose are some of the methods involving either the manipulation of membranes with transferred proteins or just a passive transfer of antigens to membranes. All these methods are briefly reviewed in this chapter. PMID:19378075

  13. Other notable protein blotting methods: a brief review.

    PubMed

    Kurien, Biji T; Scofield, R Hal

    2015-01-01

    Proteins have been transferred from the gel to the membrane by a variety of methods. These include vacuum blotting, centrifuge blotting, electroblotting of proteins to Teflon tape and membranes for N- and C-terminal sequence analysis, multiple tissue blotting, a two-step transfer of low- and high-molecular-weight proteins, acid electroblotting onto activated glass, membrane-array method for the detection of human intestinal bacteria in fecal samples, protein microarray using a new black cellulose nitrate support, electrotransfer using square wave alternating voltage for enhanced protein recovery, polyethylene glycol-mediated significant enhancement of the immunoblotting transfer, parallel protein chemical processing before and during western blot and the molecular scanner concept, electronic western blot of matrix-assisted laser desorption/ionization mass spectrometric-identified polypeptides from parallel processed gel-separated proteins, semidry electroblotting of peptides and proteins from acid-urea polyacrylamide gels, transfer of silver-stained proteins from polyacrylamide gels to polyvinylidene difluoride (PVDF) membranes, and the display of K(+) channel proteins on a solid nitrocellulose support for assaying toxin binding. The quantification of proteins bound to PVDF membranes by elution of CBB, clarification of immunoblots on PVDF for transmission densitometry, gold coating of nonconductive membranes before matrix-assisted laser desorption/ionization tandem mass spectrometric analysis to prevent charging effect for analysis of peptides from PVDF membranes, and a simple method for coating native polysaccharides onto nitrocellulose are some of the methods involving either the manipulation of membranes with transferred proteins or just a passive transfer of antigens to membranes. All these methods are briefly reviewed in this chapter. PMID:26044032

  14. A new multi-host species indirect ELISA using protein A/G conjugate for detection of anti-Toxoplasma gondii IgG antibodies with comparison to ELISA-IgG, agglutination assay and Western blot.

    PubMed

    Al-Adhami, Batol H; Gajadhar, Alvin A

    2014-02-24

    Toxoplasma gondii is a zoonotic protozoan parasite which can cause significant disease and losses in livestock and wild animals. It is increasingly recognized as an important foodborne pathogen in a broad range of food animals and products. Effective control strategies require rapid, reliable and cost-effective detection methods for large scale surveys and diagnostic applications in a broad range of warm-blooded animals. To overcome one or more of these shortcomings in the currently available detection methods for T. gondii infection a non-species-specific protein A/G conjugate was used in the development of an indirect ELISA (ELISA-A/G) for the detection of IgG antibodies in serum samples obtained from experimentally infected pigs. The performance of the assay was evaluated using serum samples from pigs, cats, mice and seals with known positive or negative status for T. gondii infection. Results of the ELISA-A/G obtained with pig serum samples were compared with those generated by traditional ELISA using host specific IgG conjugate (ELISA-IgG), modified agglutination test (MAT) and Western blot analysis (WB). Using protein A/G conjugate, comparative analysis of results from 77 samples obtained from T. gondii infected pigs showed excellent agreement between the ELISA-A/G and in-house ELISA-IgG (0.917 κ). Similar agreements were also observed when these samples were tested by a commercial ELISA kit (0.816 κ), MAT (0.816 κ) and WB (0.79 κ). A total of 86 serum samples obtained from cats, mice and seals experimentally infected with T. gondii and tested by the ELISA-A/G as well as MAT for the presence of anti-Toxoplasma IgG antibodies yielded Kappa value of 1.0 for cats and mice and 0.79 for seals. These results show that the ELISA-A/G is a suitable method for serological detection of T. gondii infection in multiple host species and has the potential for testing samples from a broad range of domestic, wild, and aquatic mammalian host species. Simultaneous testing

  15. Egypt western desert activity hits high gear

    SciTech Connect

    Petzet, G.A.

    1996-11-04

    Exploration in Egypt`s western desert is turning up large oil and gas reserves in several sub-basins. Though not a threat to the Gulf of Suez`s oil production dominance, the western desert`s output is climbing. And gas production is also set to rise as more pipelines are built. The paper describes the geology, Qarun field production, further exploration at Qarun and the Khalda/Obaiyed gas.

  16. 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

  17. 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

  18. 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

  19. The Use of Biotin to Demonstrate Immunohistochemistry, Western Blotting, and Dot Blots in University Practical Classes

    ERIC Educational Resources Information Center

    Millar, Thomas James; Knighton, Ronald; Chuck, Jo-Anne

    2012-01-01

    Immunological detection of proteins is an essential method to demonstrate to undergraduate biology students, however, is often difficult in resource and time poor student laboratory sessions. This method describes a failsafe method to rapidly and economically demonstrate this technique using biotinylated proteins or biotin itself as targets for…

  20. Influence of the Western Pacific teleconnection pattern on Western North Pacific tropical cyclone activity

    NASA Astrophysics Data System (ADS)

    Choi, Ki-Seon; Moon, Il-Ju

    2012-09-01

    This study analyzes the characteristics of Western North Pacific (WNP) tropical cyclone (TC) activity and large-scale environments according to the Western Pacific (WP) teleconnection pattern in summer. In the positive WP phase, an anomalous cyclone and an anomalous anticyclone develop in the low and middle latitudes of the East Asia area, respectively. As a result, southeasterlies are reinforced in the northeast area of East Asia (including Korea and Japan), which facilitates the movement of TC to this area, whereas northwesterlies are reinforced in the southwest area of East Asia (including southern China and the Indochina Peninsula) which blocks the movement of TC to that area. Due to the spatial distribution of this reinforced pressure system, TCs that develop during the positive WP phase move and turn more to the northeast of the WNP than TCs which develop during the negative WP phase. The characteristics of this TC activity during the positive WP phase are associated with the upper tropospheric jet being located farther to the northeast. TCs during the negative WP phase mainly move to the west from the Philippines toward southern China and the Indochina Peninsula. Due to the terrain effect caused by the passage of TCs in mainland China, the intensity of TCs during the negative WP phase is weaker than those during the positive WP phase.

  1. Detection of proteins on blot transfer membranes.

    PubMed

    Sasse, Joachim; Gallagher, Sean R

    2008-11-01

    Staining of blot transfer membranes permits visualization of proteins and allows the extent of transfer to be monitored. In the protocols described in this unit, proteins are stained after electroblotting from one-dimensional or two-dimensional polyacrylamide gels to blot membranes such as polyvinylidene difluoride (PVDF), nitrocellulose, or nylon membranes. Protocols are provided for the use of six general protein stains: Amido black, Coomassie blue, Ponceau S, colloidal gold, colloidal silver, and India ink. In addition, the fluorescent stains fluorescamine and IAEDANS, which covalently react with bound proteins, are described. Approximate detection limits for each nonfluorescent stain are indicated along with membrane compatibilities.

  2. Detection of proteins on blot membranes.

    PubMed

    Harper, S; Speicher, D W

    2001-05-01

    Staining of blot transfer membranes permits visualization of proteins and allows the extent of transfer to be monitored. In the protocols described in this unit, proteins are stained after electroblotting from one-dimensional or two-dimensional polyacrylamide gels to blot membranes such as polyvinylidene difluoride (PVDF), nitrocellulose, or nylon membranes. Protocols are provided for the use of six general protein stains: amido black, Coomassie blue, Ponceau S, colloidal gold, colloidal silver, and India ink. In addition, the fluorescent stains fluorescamine and IAEDANS, which covalently react with bound proteins, are described. Approximate detection limits for each nonfluorescent stain are indicated along with membrane compatibilities. PMID:18429099

  3. Detection of proteins on blot transfer membranes.

    PubMed

    Sasse, Joachim; Gallagher, Sean R

    2008-11-01

    Staining of blot transfer membranes permits visualization of proteins and allows the extent of transfer to be monitored. In the protocols described in this unit, proteins are stained after electroblotting from one-dimensional or two-dimensional polyacrylamide gels to blot membranes such as polyvinylidene difluoride (PVDF), nitrocellulose, or nylon membranes. Protocols are provided for the use of six general protein stains: Amido black, Coomassie blue, Ponceau S, colloidal gold, colloidal silver, and India ink. In addition, the fluorescent stains fluorescamine and IAEDANS, which covalently react with bound proteins, are described. Approximate detection limits for each nonfluorescent stain are indicated along with membrane compatibilities. PMID:19016450

  4. 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.

  5. Enzyme activities along a latitudinal transect in Western Siberia

    NASA Astrophysics Data System (ADS)

    Schnecker, Jörg; Wild, Birgit; Eloy Alves, Ricardo J.; Gentsch, Norman; Gittel, Antje; Knoltsch, Anna; Lashchinskiy, Nikolay; Mikutta, Robert; Takriti, Mounir; Richter, Andreas

    2014-05-01

    Decomposition of soil organic matter (SOM) and thus carbon and nutrient cycling in soils is mediated by the activity of extracellular enzymes. The specific activities of these enzymes and their ratios to each other represent the link between the composition of soil organic matter and the nutrient demand of the microbial community. Depending on the difference between microbial nutrient demand and substrate availability, extracellular enzymes can enhance or slow down different nutrient cycles in the soil. We investigated activities of six extracellular enzymes (cellobiohydrolase, leucine-amino-peptidase, N-acetylglucosaminidase, chitotriosidase, phosphatase and phenoloxidase) in the topsoil organic horizon, topsoil mineral horizon and subsoil horizon in seven ecosystems along a 1,500 km-long North-South transect in Western Siberia. The transect included sites in the southern tundra, northern taiga, middle taiga, southern taiga, forest-steppe (in forested patches as well as in adjacent meadows) and Steppe. We found that enzyme patterns varied stronger with soil depth than between ecosystems. Differences between horizons were mainly based on the increasing ratio of oxidative enzymes to hydrolytic enzymes. Differences between sites were more pronounced in topsoil than in subsoil mineral horizons, but did not reflect the north-south transect and the related gradients in temperature and precipitation. The observed differences between sites in topsoil horizons might therefore result from differences in vegetation rather than climatic factors. The decreasing variability in the enzyme pattern with depth might also indicate that the composition of soil organic matter becomes more similar with soil depth, most likely by an increasing proportion of microbial remains compared to plant derived constituents of SOM. This also indicates, that SOM becomes less divers the more it is processed by soil microorganisms. Our findings highlight the importance of soil depth on enzyme

  6. 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

  7. 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.

  8. Antifungal activity of extracts and select compounds in heartwood of seven western conifers toward Phytophthora ramorum

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Individual compounds and ethyl acetate extracts from heartwood of seven conifer species were tested for fungicidal activity against Phytophthora ramorum. Extracts from incense and western red cedar exhibited the strongest activity (EC50 589 and 646 ppm, respectively), yellow-cedar, western juniper,...

  9. Western lowland gorillas (Gorilla gorilla gorilla) change their activity patterns in response to frugivory.

    PubMed

    Masi, Shelly; Cipolletta, Chloé; Robbins, Martha M

    2009-02-01

    The most important environmental factor explaining interspecies variation in ecology and sociality of the great apes is likely to be variation in resource availability. Relatively little is known about the activity patterns of western lowland gorillas (Gorilla gorilla gorilla), which inhabit a dramatically different environment from the well-studied mountain gorillas (G. beringei beringei). This study aims to provide a detailed quantification of western lowland gorillas' activity budgets using direct observations on one habituated group in Bai Hokou, Central African Republic. We examined how activity patterns of both sexes are shaped by seasonal frugivory. Activity was recorded with 5-min instantaneous sampling between December 2004 and December 2005. During the high-frugivory period the gorillas spent less time feeding and more time traveling than during the low-frugivory period. The silverback spent less time feeding but more time resting than both females and immatures, which likely results from a combination of social and physiological factors. When compared with mountain gorillas, western lowland gorillas spend more time feeding (67 vs. 55%) and traveling (12 vs. 6.5%), but less time resting (21 vs. 34%) and engaging in social/other activities (0.5 vs. 3.6%). This disparity in activity budgets of western lowland gorillas and mountain gorillas may be explained by the more frugivorous diet and the greater dispersion of food resources experienced by western lowland gorillas. Like other apes, western lowland gorillas change their activity patterns in response to changes in the diet.

  10. Active population growth and immigration hypotheses in Western Europe.

    PubMed

    Feld, S

    2000-03-01

    The paper examines, in respect of 12 Western European countries over a period of 20 years, the widely held view that any decline in their working population should be offset by greater reliance on immigrant labor. This research, based on demographic projections and forecasts regarding labor market participation rates by age and sex for each of the countries concerned, focuses on the two most likely scenarios. It appears that only Italy will be faced with a fall in its working population. All other western countries will either maintain the same level or, more generally, see their workforce grow substantially. Accordingly, the authors may safely assert that there is no risk of a shortage of workers between now and the year 2020, and that an increasing supply of labor will render reliance on a greater influx of immigrant workers unnecessary. The second part analyses changes in the structure of the demand for labor. The authors deal chiefly with the phenomenon of the concentration of foreign manpower in each sector, its flexibility and mobility in a context of unemployment, as well as the impact of new technologies and globalization on the main determinants of international migration of labor.

  11. Active deformation in Western Turkey: new GPS observations and models

    NASA Astrophysics Data System (ADS)

    Nocquet, J.; Aktug, B.; Parsons, B.; Cingoz, A.; England, P.; Erkan, Y.; Soyer, N.; Akdeniz, H.; Kilicoglu, A.

    2007-12-01

    How the continents deform remains a matter of debate. One view postulates that continental deforming zones are comprised of a limited numbers of rigid (elastic) microplates. If true, the surface motion can then be described by the relative rotation of blocks, and strain should be localized along the major faults separating the blocks. An alternative view is that the deformation at depth is distributed over wide areas, can be modelled by a viscous flow responding to boundary conditions applied on it and gravitational potential energy gradients related to variations in topography, and the surface strain simply reflects this deformation. Western Turkey is a region of crustal extension, part of the Nubia/Eurasia plate boundary. Its kinematics is often modelled by the relative motion of a small number of rigid blocks (Nyst & Thatcher, 2005, Reilinger et al., 2006). However, until now, the limited number of GPS velocity vectors available has prevented a detailed examination of which is the more appropriate description. We present a new geodetic velocity field including ~100 sites from the longitude the Central Anatolian plateau to the Aegean coast, derived from a combination of campaigns carried out between 1997 and 2006, and continuous GPS operating since 2003, which we use to test the different models. While the kinematics of the area can be correctly modelled by a block model, a good fit to the velocity field requires blocks with sizes smaller than 100 km and still fails to adequately predict the strain rate observed within blocks . Alternatively, we test an approach where the lithosphere is modelled as a thin viscous sheet, responding to the gravitational potentiel energy contrast between the high plateau of eastern Turkey to the east and the subduction along the Hellenic trench in the southwest. The simplistic model has only one free parameter (the force applied by the subducting oceanic lithosphere on the Aegean ), but provides a good agreement with the observed

  12. Electrospun nitrocellulose and nylon: Design and fabrication of novel high performance platforms for protein blotting applications

    PubMed Central

    Manis, Ashley E; Bowman, James R; Bowlin, Gary L; Simpson, David G

    2007-01-01

    Background Electrospinning is a non-mechanical processing strategy that can be used to process a variety of native and synthetic polymers into highly porous materials composed of nano-scale to micron-scale diameter fibers. By nature, electrospun materials exhibit an extensive surface area and highly interconnected pore spaces. In this study we adopted a biological engineering approach to ask how the specific unique advantages of the electrospinning process might be exploited to produce a new class of research/diagnostic tools. Methods The electrospinning properties of nitrocellulose, charged nylon and blends of these materials are characterized. Results Nitrocellulose electrospun from a starting concentration of < 110 mg/ml acetone deposited as 4–8 μm diameter beads; at 110 mg/ml-to-140 mg/ml starting concentrations, this polymer deposited as 100–4000 nm diameter fibers. Nylon formed fibers when electrospun from 60–140 mg/ml HFIP, fibers ranged from 120 nm-6000 nm in diameter. Electrospun nitrocellulose exhibited superior protein retention and increased sensitivity in slot blot experiments with respect to the parent nitrocellulose material. Western immunoblot experiments using fibronectin as a model protein demonstrated that electrospun nylon exhibits increased protein binding and increased dynamic range in the chemiluminescence detection of antigens than sheets of the parent starting material. Composites of electrospun nitrocellulose and electrospun nylon exhibit high protein binding activity and provide increased sensitivity for the immuno-detection of antigens. Conclusion The flexibility afforded by electrospinning process makes it possible to tailor blotting membranes to specific applications. Electrospinning has a variety of potential applications in the clinical diagnostic field of use. PMID:18271978

  13. A bead-based western for high-throughput cellular signal transduction analyses

    PubMed Central

    Treindl, Fridolin; Ruprecht, Benjamin; Beiter, Yvonne; Schultz, Silke; Döttinger, Anette; Staebler, Annette; Joos, Thomas O.; Kling, Simon; Poetz, Oliver; Fehm, Tanja; Neubauer, Hans; Kuster, Bernhard; Templin, Markus F.

    2016-01-01

    Dissecting cellular signalling requires the analysis of large number of proteins. The DigiWest approach we describe here transfers the western blot to a bead-based microarray platform. By combining gel-based protein separation with immobilization on microspheres, hundreds of replicas of the initial blot are created, thus enabling the comprehensive analysis of limited material, such as cells collected by laser capture microdissection, and extending traditional western blotting to reach proteomic scales. The combination of molecular weight resolution, sensitivity and signal linearity on an automated platform enables the rapid quantification of hundreds of specific proteins and protein modifications in complex samples. This high-throughput western blot approach allowed us to identify and characterize alterations in cellular signal transduction that occur during the development of resistance to the kinase inhibitor Lapatinib, revealing major changes in the activation state of Ephrin-mediated signalling and a central role for p53-controlled processes. PMID:27659302

  14. Warming and earlier spring increase Western U.S. forest wildfire activity

    USGS Publications Warehouse

    Westerling, A.L.; Hidalgo, H.G.; Cayan, D.R.; Swetnam, T.W.

    2006-01-01

    Western United States forest wildfire activity is widely thought to have increased in recent decades, yet neither the extent of recent changes nor the degree to which climate may be driving regional changes in wildfire has been systematically documented. Much of the public and scientific discussion of changes in western United States wildfire has focused instead on the effects of 19th- and 20th-century land-use history. We compiled a comprehensive database of large wildfires in western United States forests since 1970 and compared it with hydroclimatic and land-surface data. Here, we show that large wildfire activity increased suddenly and markedly in the mid-1980s, with higher large-wildfire frequency, longer wildfire durations, and longer wildfire seasons. The greatest increases occurred in mid-elevation, Northern Rockies forests, where land-use histories have relatively little effect on fire risks and are strongly associated with increased spring and summer temperatures and an earlier spring snowmelt.

  15. 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

  16. Validation of a Dot-Blot quantitative technique for large scale analysis of beef tenderness biomarkers.

    PubMed

    Guillemin, N; Meunier, B; Jurie, C; Cassar-Malek, I; Hocquette, J-F; Leveziel, H; Picard, B

    2009-10-01

    Beef tenderness is a very complex and multifactorial sensorial meat quality trait, which depends partly on muscle characteristics. This tissue is very variable according to animal type (age, breed and sex) and rearing conditions. Consequently, beef tenderness exhibits a great variability. Different research programs have revealed several genes or proteins which could be good markers of beef tenderness. In order to validate the relation of these markers with beef tenderness on a large population of bovines, it is necessary to have a large-scale and trusty technique which can access different quantities of proteins related to tenderness. In this study we firstly compared Western-Blot and Dot-Blot. Secondly, we evaluated Dot-Blot technical and biological capabilities for the quantification of protein biomarkers. The results demonstrated that the Dot-Blot technique with fluorescence detection presents numerous interests. This technique allows a good reproducibility and permits the simultaneous analysis of a large number of samples. The Dot-Blot technique defined and validated in this study can be used for protein biomarkers analyses, notably to predict beef tenderness. Another major result of this study is that about 5 to 10 animals per group are required to detect large differences (>1.5) in biomarker expression between tender and tough beef, whereas much larger numbers of animals (10 to 30) are required to detect smaller differences (about 1.2 to 1.3) taking into account the biological variability of these markers.

  17. Controls on interannual variability in lightning-caused fire activity in the western US

    NASA Astrophysics Data System (ADS)

    Abatzoglou, John T.; Kolden, Crystal A.; Balch, Jennifer K.; Bradley, Bethany A.

    2016-04-01

    Lightning-caused wildfires account for a majority of burned area across the western United States (US), yet lightning remains among the more unpredictable spatiotemporal aspects of the fire environment and a challenge for both modeling and managing fire activity. A data synthesis of cloud-to-ground lightning strikes, climate and fire data across the western US from 1992 to 2013 was conducted to better understand geographic variability in lightning-caused wildfire and the factors that influence interannual variability in lightning-caused wildfire at regional scales. Distinct geographic variability occurred in the proportion of fires and area burned attributed to lightning, with a majority of fires in the interior western US attributed to lightning. Lightning ignition efficiency was highest across the western portion of the region due to the concomitance of peak lightning frequency and annual nadir in fuel moisture in mid-to-late summer. For most regions the number of total and dry lightning strikes exhibited strong interannual correlation with the number of lightning-caused fires, yet were a poor predictor of area burned at regional scales. Commonality in climate-fire relationships for regional annual area burned by lightning- versus human-ignited fires suggests climate conditions, rather than lightning activity, are the predominant control of interannual variability in area burned by lightning-caused fire across much of the western US.

  18. 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

  19. Improvement of antigen blotting in a tissue blot immunobinding assay for the detection of two chili pepper viruses.

    PubMed

    Han, Jung-Heon; Shin, Jun-Sung; Kim, Young Ho; Kim, Byung-Dong

    2007-11-01

    The tissue blot immunobinding assay (TBIA) is widely used for the detection and localization of plant viruses in various plant tissues. The basic experimental procedures of TBIA sampling and blotting were simplified using commercially available micropipette tips. This method was termed the ring-blot immunobinding assay (R-BIA), as the blot on the membrane forms a ring shape. The detection efficacy of R-BIA was tested for two chili pepper viruses, pepper mild mottle tobamovirus (PMMoV) and pepper mottle potyvirus (PepMoV), following the optimized serological procedures of TBIA (length of the incubation period and BSA concentration, and primary and secondary antibodies). Sensitivity of the R-BIA was about 1 ng/ml of purified PMMoV in pepper leaf sap from a healthy pepper plant. R-BIA also showed high specificity in the detection of PMMoV and PepMoV. Moreover, the modified sampling and blotting procedures were simpler and more reliable than other TBIA methods (such as whole-leaf blotting and crushed-leaf blotting), suggesting that the R-BIA may be used for medium- to large-scale detection of plant viruses in laboratories with minimal facilities.

  20. Transcription Factor Proteomics: Identification by a Novel Gel Mobility Shift-Three-Dimensional Electrophoresis Method Coupled with Southwestern Blot and HPLC-Electrospray-Mass Spectrometry Analysis

    PubMed Central

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

    2011-01-01

    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 ×107 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

  1. Preliminary cellular-automata forecast of permit activity from 1998 to 2010, Idaho and Western Montana

    USGS Publications Warehouse

    Raines, G.L.; Zientek, M.L.; Causey, J.D.; Boleneus, D.E.

    2002-01-01

    For public land management in Idaho and western Montana, the U.S. Forest Service (USFS) has requested that the U.S. Geological Survey (USGS) predict where mineral-related activity will occur in the next decade. Cellular automata provide an approach to simulation of this human activity. Cellular automata (CA) are defined by an array of cells, which evolve by a simple transition rule, the automaton. Based on exploration trends, we assume that future exploration will focus in areas of past exploration. Spatial-temporal information about mineral-related activity, that is permits issued by USFS and Bureau of Land Management (BLM) in the last decade, and spatial information about undiscovered resources, provide a basis to calibrate a CA. The CA implemented is a modified annealed voting rule that simulates mineral-related activity with spatial and temporal resolution of 1 mi2 and 1 year based on activity from 1989 to 1998. For this CA, the state of the economy and exploration technology is assumed constant for the next decade. The calibrated CA reproduces the 1989-1998-permit activity with an agreement of 94%, which increases to 98% within one year. Analysis of the confusion matrix and kappa correlation statistics indicates that the CA underestimates high activity and overestimates low activity. Spatially, the major differences between the actual and calculated activity are that the calculated activity occurs in a slightly larger number of small patches and is slightly more uneven than the actual activity. Using the calibrated CA in a Monte Carlo simulation projecting from 1998 to 2010, an estimate of the probability of mineral activity shows high levels of activity in Boise, Caribou, Elmore, Lincoln, and western Valley counties in Idaho and Beaverhead, Madison, and Stillwater counties in Montana, and generally low activity elsewhere. ?? 2002 International Association for Mathematical Geology.

  2. Mixed lubrication after rewetting of blotted pleural mesothelium.

    PubMed

    Bodega, Francesca; Sironi, Chiara; Porta, Cristina; Pecchiari, Matteo; Zocchi, Luciano; Agostoni, Emilio

    2013-01-15

    Coefficient of kinetic friction (μ) of pleural mesothelium blotted with filter paper, and rewetted with Ringer solution markedly increases; this increase is removed if a sufficient amount of sialomucin or hyaluronan is added to Ringer (Bodega et al., 2012. Respiratory Physiology and Neurobiology 180, 34-39). In this research we found that μ of pleural mesothelium blotted, rewetted, and sliding at physiological velocities and loads, decreased with increase of velocity, mainly at low velocities. Despite this decrease, μ at highest velocity was still double that before blotting. With small concentration of sialomucin or hyaluronan μ was markedly smaller at each velocity, decreased less with increase of velocity, and at highest velocity approached preblotting value. These findings indicate a regime of mixed lubrication in post-blotting Ringer, at variance with boundary lubrication occurring before blotting or postblotting with sufficient macromolecule addition. Greater roughness of mesothelial surface, caused by blotting, likely induces zones of elastohydrodynamic lubrication, which increase with velocity, while contact area decreases.

  3. EarthScope Transportable Array Siting Outreach Activities in Alaska and Western Canada

    NASA Astrophysics Data System (ADS)

    Dorr, P. M.; Gardine, L.; Tape, C.; McQuillan, P.; Cubley, J. F.; Samolczyk, M. A.; Taber, J.; West, M. E.; Busby, R.

    2015-12-01

    The EarthScope Transportable Array is deploying about 260 stations in Alaska and western Canada. IRIS and EarthScope are partnering with the Alaska Earthquake Center, part of the University of Alaska's Geophysical Institute, and Yukon College to spread awareness of earthquakes in Alaska and western Canada and the benefits of the Transportable Array for people living in these regions. We provide an update of ongoing education and outreach activities in Alaska and Canada as well as continued efforts to publicize the Transportable Array in the Lower 48. Nearly all parts of Alaska and portions of western Canada are tectonically active. The tectonic and seismic variability of Alaska, in particular, requires focused attention at the regional level, and the remoteness and inaccessibility of most Alaskan and western Canadian villages and towns often makes frequent visits difficult. When a community is accessible, every opportunity to engage the residents is made. Booths at state fairs and large cultural gatherings, such as the annual convention of the Alaska Federation of Natives, are excellent venues to distribute earthquake information and to demonstrate a wide variety of educational products and web-based applications related to seismology and the Transportable Array that residents can use in their own communities. Meetings and interviews with Alaska Native Elders and tribal councils discussing past earthquakes has led to a better understanding of how Alaskans view and understand earthquakes. Region-specific publications have been developed to tie in a sense of place for residents of Alaska and the Yukon. The Alaska content for IRIS's Active Earth Monitor emphasizes the widespread tectonic and seismic features and offers not just Alaska residents, but anyone interested in Alaska, a glimpse into what is going on beneath their feet. The concerted efforts of the outreach team will have lasting effects on Alaskan and Canadian understanding of the seismic hazard and

  4. From globalization to Europeanization-and then? Transnational influences in lesbian activism of the Western Balkans.

    PubMed

    Dioli, Irene

    2011-01-01

    The article looks at transnational dynamics in lesbian activism in the Western Balkans, with particular focus on Serbia and Bosnia and Herzegovina, over the last two decades. Based on 30 interviews with activists from local and international feminist, lesbian, gay, bisexual, and transgender (LGBT), and queer organizations, the article discusses the role played by transnational contacts in the development of lesbian activism in the region and the related issues arising within the conflicting relationship between the "East" and the "West," with special attention to trends related to international intervention and cooperation, globalization, and Europeanization. PMID:21774599

  5. Implications of perennial saline springs for abnormally high fluid pressures and active thrusting in western California

    SciTech Connect

    Unruh, J.R.; Davisson, M.L.; Criss, R.E.; Moores, E.M. )

    1992-05-01

    Perennial saline springs in the Rumsey Hills area, southwestern Sacramento Valley, California, locally discharge at high elevations and near ridgetops. The springs are cold, are commonly associated with natural gas seeps, and typically emerge along west-vergent thrust faults. Stable isotope analyses indicate that the spring waters are similar to oil-field formation fluids and they have had a significant residence time in the subsurface at moderate temperatures. The nonmeteoric character of the springs demonstrates that they are not being fed by perched water tables. The authors propose that these subsurface formation waters are being forced to the surface by anomalously high porefluid pressures. The Rumsey Hills area is one of Quaternary uplift, thrusting, and crustal shortening, and prospect wells drilled there have encountered anomalously high fluid pressures at shallow depths. They attribute these high fluid pressures to active tectonic compression and shortening of Cretaceous marine sedimentary rocks. The widespread occurrence of anomalously high pore-fluid pressures and perennial saline springs in the Coast Ranges and western Great Valley suggests that much of western California may be characterized as a seismically active, overpressured thrust belt. The emergence of formation waters along thrust faults further suggests that patterns of subsurface fluid flow in western California may be similar to those in overpressured accretionary prisms, and that excess fluid pressures may also play a role in the distribution of seismicity.

  6. Evaluating amphibian responses in wetlands impacted by mining activities in the western United States

    SciTech Connect

    Linder, G.; Wyant, J.; Meganck, R.; Williams, B.

    1991-01-01

    An increasing awareness of declining amphibian populations in the United States requires that the authors develop strategies for evaluating anthropogenic impacts on wetlands and the biota dependent upon these habitats. For example, in the western United States, mining activities may impact a wetland and its biota directly through habitat destruction or run-off of sediments and contaminants generated during mining operations. Amphibians which frequent these transition zones between terrestrial and aquatic habitats may be key biological indicators of a wetland's status. Through a demonstration project located in the mining regions of western Montana, the authors are currently using laboratory and field methods for a wetland evaluation required within a Superfund ecological risk assessment.

  7. Palaeoseismological evidence for Holocene activity on the Manisa Fault Zone,Western Anatolia

    NASA Astrophysics Data System (ADS)

    Özkaymak, Ç.; Sözbilir, H.; Uzel, B.; Akyüz, H. S.

    2009-04-01

    Manisa Fault Zone (MFZ) is an active structural discontinuity that is geomorphologically expressed as a trace of north-facing Quaternary fault scarps bounding the southern margin of the Manisa basin which is subsidiary to the Gediz Graben. We note that the present-day fault trace is over 50 km long from Manisa city in the northwest to the Turgutlu town in the southeast. The MFZ consists of two major sections: (i) eastern section that strikes NW-SE direction in the south and bends into an approximately E-W direction around Manisa to the northwest, (ii) an approximately 10-km-long western section that strikes approximately WNW-ESE direction from Manisa city in the east to the Akgedik town in the west. In this study, we present the geologic, geomorphologic, and palaeoseismologic observations indicating Holocene activity on the western section of the fault zone. We identify that the MFZ, at its western end, consists of three fault segments which are en échelon arranged in left step; the fault segments show evidence for linkage and breaching at the relay ramps. One of them is named as the Manastir Fault. In front of this fault, two Holocene colluvial fans older of which is uncorformity bounded are cut and displaced by the syntethic faults. Palaeoseismologic data show that the syntethic fault segments correspond to the surface ruptures of the historical earthquakes. As a result of detailed stratigraphic, sedimentologic and structural observations on the trench walls, some evidences for at least two earthquakes are recorded which are supported by radio-carbon dating. Besides this, an archaic aqueduct that were used to transport water from Emlakdere town, located on the hanging wall of the Manastir Fault, to the basin is cut and displaced by the syntethic fault egments. It is known that this archaic architecture were in use after 11. century by the Ottomans. On the basis of the mentioned data, fault segments which are belong to the western part of the Manisa Fault Zone

  8. 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

  9. 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

  10. ASBESTOS-INDUCED ACTIVATION OF CELL SIGNALING PATHWAYS IN HUMAN BRONCHIAL EPITHELIAL CELLS

    EPA Science Inventory

    Using respiratory epithelial cells transfected with either superoxide dismutase (SOD) or catalase, the authors tested the hypothesis that the activation of the epidermal growth factor (EGF) receptor signal pathway after asbestos exposure involves an oxidative stress. Western blot...

  11. Significance of an Active Volcanic Front in the Far Western Aleutian Arc

    NASA Astrophysics Data System (ADS)

    Yogodzinski, G. M.; Kelemen, P. B.; Hoernle, K.

    2015-12-01

    Discovery of a volcanic front west of Buldir Volcano, the western-most emergent Aleutian volcano, demonstrates that the surface expression of Aleutian volcanism falls below sea level just west of 175.9° E longitude, but is otherwise continuous from mainland Alaska to Kamchatka. The newly discovered sites of western Aleutian seafloor volcanism are the Ingenstrem Depression, a 60 km-long structural depression just west of Buldir, and an unnamed area 300 km further west, referred to as the Western Cones. These locations fall along a volcanic front that stretches from Buldir to Piip Seamount near the Komandorsky Islands. Western Aleutian seafloor volcanic rocks include large quantities of high-silica andesite and dacite, which define a highly calc-alkaline igneous series and carry trace element signatures that are unmistakably subduction-related. This indicates that subducting oceanic lithosphere is present beneath the westernmost Aleutian arc. The rarity of earthquakes below depths of 200 km indicates that the subducting plate is unusually hot. Some seafloor volcanoes are 6-8 km wide at the base, and so are as large as many emergent Aleutian volcanoes. The seafloor volcanoes are submerged in water depths >3000 m because they sit on oceanic lithosphere of the Bering Sea. The volcanic front is thus displaced to the north of the ridge of arc crust that underlies the western Aleutian Islands. This displacement, which developed since approximately 6 Ma when volcanism was last active on the islands, must be a consequence of oblique convergence in a system where the subducting plate and large blocks of arc crust are both moving primarily in an arc-parallel sense. The result is a hot-slab system where low subduction rates probably limit advection of hot mantle to the subarc, and produce a relatively cool and perhaps stagnant mantle wedge. The oceanic setting and highly oblique subduction geometry also severely limit rates of sediment subduction, so the volcanic rocks, which

  12. Eocene activity on the Western Sierra Fault System and its role incising Kings Canyon, California

    NASA Astrophysics Data System (ADS)

    Sousa, Francis J.; Farley, Kenneth A.; Saleeby, Jason; Clark, Marin

    2016-04-01

    Combining new and published apatite (U-Th)/He and apatite 4He/3He data from along the Kings River canyon, California we rediscover a west-down normal fault on the western slope of the southern Sierra Nevada, one of a series of scarps initially described by Hake (1928) which we call the Western Sierra Fault System. Integrating field observations with apatite (U-Th)/He data, we infer a single fault trace 30 km long, and constrain the vertical offset across this fault to be roughly a kilometer. Thermal modeling of apatite 4He/3He data documents a pulse of footwall cooling near the fault and upstream in the footwall at circa 45-40 Ma, which we infer to be the timing of a kilometer-scale incision pulse resulting from the fault activity. In the context of published data from the subsurface of the Sacramento and San Joaquin Valleys, our data from the Western Sierra Fault System suggests an Eocene tectonic regime dominated by low-to-moderate magnitude extension, surface uplift, and internal structural deformation of the southern Sierra Nevada and proximal Great Valley forearc.

  13. Sediment yield from the tectonically active semiarid Western Transverse Ranges of California

    USGS Publications Warehouse

    Warrick, J.A.; Mertes, L.A.K.

    2009-01-01

    Sediment yields from the world's rivers are generally highest from steep drainage basins with weak lithology, active tectonics, or severe land-use impacts. Here, we evaluate sediment yields from the Western Transverse Ranges of California in an attempt to explain why they are two- to tenfold greater than the surrounding areas of California. We found that suspended-sediment yields across the gauged basins of the Western Transverse Range during 1969-1999 varied by approximately an order of magnitude (740-5300 t/km2/yr). Similarly, fine-sediment concentrations for normalized discharge rates varied by almost two orders of magnitude (e.g., 1.3-110 g/L for the mean annual flood) for 11 previously unmonitored drainages of the Santa Ynez Mountains. Areas with high sediment yields consistently have weakly consolidated bedrock (Quaternary-Pliocene marine formations) and are associated with the highest rates of tectonic uplift of the region (>5 mm/yr). These regions are important to the sediment discharge budgets, because ???50% of the total suspended-sediment discharge from the Western Transverse Range is estimated to be generated within these regions, even though they represent only ???10% of the total watershed area. Previous estimates of suspended-sediment discharge from the Ventura River have likely been underestimated by ???50% because the gauging station is located immediately upstream of a high sediment yield region. We also found a significant and positive correlation between sediment yield and the percentage of a watershed with grassland and agricultural land use. These results suggest that there is adequate variation within the lithology, tectonics, and land use of the broader Western Transverse Range geologic province to induce large variations in sediment yield at the local scale. ?? 2009 Geological Society of America.

  14. The effect of mayfly (Hexagenia spp.) burrowing activity on sediment oxygen demand in western Lake Erie

    USGS Publications Warehouse

    Edwards, William J.; Soster, Frederick M.; Matisoff, Gerald; Schloesser, Donald W.

    2009-01-01

    Previous studies support the hypothesis that large numbers of infaunal burrow-irrigating organisms in the western basin of Lake Erie may increase significantly the sediment oxygen demand, thus enhancing the rate of hypolimnetic oxygen depletion. We conducted laboratory experiments to quantify burrow oxygen dynamics and increased oxygen demand resulting from burrow irrigation using two different year classes of Hexagenia spp. nymphs from western Lake Erie during summer, 2006. Using oxygen microelectrodes and hot film anemometry, we simultaneously determined oxygen concentrations and burrow water flow velocities. Burrow oxygen depletion rates ranged from 21.7 mg/nymph/mo for 15 mm nymphs at 23 °C to 240.7 mg/nymph/mo for 23 mm nymphs at 13 °C. Sealed microcosm experiments demonstrated that mayflies increase the rate of oxygen depletion by 2-5 times that of controls, depending on size of nymph and water temperature, with colder waters having greater impact. At natural population densities, nymph pumping activity increased total sediment oxygen demand 0.3-2.5 times compared to sediments with no mayflies and accounted for 22-71% of the total sediment oxygen demand. Extrapolating laboratory results to the natural system suggest that Hexagenia spp. populations may exert a significant control on oxygen depletion during intermittent stratification. This finding may help explain some of the fluctuations in Hexagenia spp. population densities in western Lake Erie and suggests that mayflies, by causing their own population collapse irrespective of other environmental conditions, may need longer term averages when used as a bio-indicator of the success of pollution-abatement programs in western Lake Erie and possibly throughout the Great Lakes.

  15. The use of circulation weather types to predict upwelling activity along the Western Iberian Peninsula coast

    NASA Astrophysics Data System (ADS)

    Ramos, Alexandre M.; Cordeiro Pires, Ana; Sousa, Pedro M.; Trigo, Ricardo M.

    2013-04-01

    Coastal upwelling is a phenomenon that occurs in most western oceanic coasts due to the presence of mid-latitude high-pressure systems that generate equatorward winds along the coast and consequent offshore displacement of surface waters that in turn cause deeper, colder, nutrient-rich waters to arise. In western Iberian Peninsula (IP) the high-pressure system associated to northerly winds occurs mainly during spring and summer. Upwelling systems are economically relevant, being the most productive regions of the world ocean and crucial for fisheries. In this work, we evaluate the intra- and inter-annual variability of the Upwelling Index (UI) off the western coast of the IP considering four locations at various latitudes: Rias Baixas, Aveiro, Figueira da Foz and Cabo da Roca. In addition, the relationship between the variability of the occurrence of several circulation weather types (Ramos et al., 2011) and the UI variability along this coast was assessed in detail, allowing to discriminate which types are frequently associated with strong and weak upwelling activity. It is shown that upwelling activity is mostly driven by wind flow from the northern quadrant, for which the obtained correlation coefficients (for the N and NE types) are higher than 0.5 for the four considered test locations. Taking into account these significant relationships, we then developed statistical multi-linear regression models to hindcast upwelling series (April to September) at the four referred locations, using monthly frequencies of circulation weather types as predictors. Modelled monthly series reproduce quite accurately observational data, with correlation coefficients above 0.7 for all locations, and relatively small absolute errors. Ramos AM, Ramos R, Sousa P, Trigo RM, Janeira M, Prior V (2011) Cloud to ground lightning activity over Portugal and its association with Circulation Weather Types. Atmospheric Research 101:84-101. doi: 10.1016/j.atmosres.2011.01

  16. MULTIPLE EPISODES OF IGNEOUS ACTIVITY, MINERALIZATION, AND ALTERATION IN THE WESTERN TUSHAR MOUNTAINS, UTAH.

    USGS Publications Warehouse

    Cunningham, Charles G.; Steven, Thomas A.; Campbell, David L.; Naeser, Charles W.; Pitkin, James A.; Duval, Joseph S.

    1984-01-01

    The report outlines the complex history of igneous activity and associated alteration and mineralization in the western Tushar Mountains, Utah and pointss out implciations for minerals exploration. The area has been subjected to recurrent episodes of igneous intrusion, hydrothermal alteration, and mineralization, and the mineral-resource potential of the different mineralized areas is directly related to local geologic history. The mineral commodities to be expected vary from one hydrothermal system to another, and from one depth to another within any given system. Uranium and molybdenum seem likely to have the greatest economic potential, although significant concentrations of gold may also exist.

  17. Modulation of western North Pacific tropical cyclone activity by the Atlantic Meridional Mode

    NASA Astrophysics Data System (ADS)

    Zhang, Wei; Vecchi, Gabriel A.; Villarini, Gabriele; Murakami, Hiroyuki; Rosati, Anthony; Yang, Xiaosong; Jia, Liwei; Zeng, Fanrong

    2016-05-01

    This study examines the year-to-year modulation of the western North Pacific (WNP) tropical cyclones (TC) activity by the Atlantic Meridional Mode (AMM) using both observations and the Geophysical Fluid Dynamics Laboratory Forecast-oriented Low Ocean Resolution Version of CM2.5 (FLOR) global coupled model. 1. The positive (negative) AMM phase suppresses (enhances) WNP TC activity in observations. The anomalous occurrence of WNP TCs results mainly from changes in TC genesis in the southeastern part of the WNP. 2. The observed responses of WNP TC activity to the AMM are connected to the anomalous zonal vertical wind shear (ZVWS) caused by AMM-induced changes to the Walker circulation. During the positive AMM phase, the warming in the North Atlantic induces strong descending flow in the tropical eastern and central Pacific, which intensifies the Walker cell in the WNP. The intensified Walker cell is responsible for the suppressed (enhanced) TC genesis in the eastern (western) part of the WNP by strengthening (weakening) ZVWS. 3. The observed WNPTC-AMM linkage is examined by the long-term control and idealized perturbations experiment with FLOR-FA. A suite of sensitivity experiments strongly corroborate the observed WNPTC-AMM linkage and underlying physical mechanisms.

  18. Seasonal prediction of lightning activity in North Western Venezuela: Large-scale versus local drivers

    NASA Astrophysics Data System (ADS)

    Muñoz, Á. G.; Díaz-Lobatón, J.; Chourio, X.; Stock, M. J.

    2016-05-01

    The Lake Maracaibo Basin in North Western Venezuela has the highest annual lightning rate of any place in the world (~ 200 fl km- 2 yr- 1), whose electrical discharges occasionally impact human and animal lives (e.g., cattle) and frequently affect economic activities like oil and natural gas exploitation. Lightning activity is so common in this region that it has a proper name: Catatumbo Lightning (plural). Although short-term lightning forecasts are now common in different parts of the world, to the best of the authors' knowledge, seasonal prediction of lightning activity is still non-existent. This research discusses the relative role of both large-scale and local climate drivers as modulators of lightning activity in the region, and presents a formal predictability study at seasonal scale. Analysis of the Catatumbo Lightning Regional Mode, defined in terms of the second Empirical Orthogonal Function of monthly Lightning Imaging Sensor (LIS-TRMM) and Optical Transient Detector (OTD) satellite data for North Western South America, permits the identification of potential predictors at seasonal scale via a Canonical Correlation Analysis. Lightning activity in North Western Venezuela responds to well defined sea-surface temperature patterns (e.g., El Niño-Southern Oscillation, Atlantic Meridional Mode) and changes in the low-level meridional wind field that are associated with the Inter-Tropical Convergence Zone migrations, the Caribbean Low Level Jet and tropical cyclone activity, but it is also linked to local drivers like convection triggered by the topographic configuration and the effect of the Maracaibo Basin Nocturnal Low Level Jet. The analysis indicates that at seasonal scale the relative contribution of the large-scale drivers is more important than the local (basin-wide) ones, due to the synoptic control imposed by the former. Furthermore, meridional CAPE transport at 925 mb is identified as the best potential predictor for lightning activity in the Lake

  19. Influence of human activity patterns on epidemiology of plague in Western Usambara Mountains, Tanzania.

    PubMed

    Hubeau, Marianne; Gulinck, Hubert; Kimaro, Didas N; Hieronimo, Proches; Meliyo, Joel

    2014-07-01

    Human plague has been a recurring public health threat in some villages in the Western Usambara Mountains, Tanzania, in the period between 1980 and 2004. Despite intensive past biological and medical research, the reasons for the plague outbreaks in the same set of villages remain unknown. Plague research needs to broaden its scope and formulate new hypotheses. This study was carried out to establish relationships between the nature and the spatial extent of selected human activities on one hand, and the reported plague cases on the other hand. Three outdoor activities namely, fetching water, collecting firewood and going to the market, were selected. Through enquiries the activity patterns related to these activities were mapped in 14 villages. Standard deviation ellipses represent the extent of action spaces. Over 130 activity types were identified and listed. Of these, fetching water, collecting firewood and going to the market were used for further analysis. The results indicate a significant correlation between the plague frequency and the size of these action spaces. Different characteristics of land use and related human activities were correlated with the plague frequency at village and hamlet levels. Significant relationships were found between plague frequency and specific sources of firewood and water, and specific market places.

  20. Mammalian. cap alpha. -polymerase: cloning of partial complementary DNA and immunobinding of catalytic subunit in crude homogenate protein blots

    SciTech Connect

    SenGupta, D.N.; Kumar, P.; Zmudzka, B.Z.; Coughlin, S.; Vishwanatha, J.K.; Robey, F.A.; Parrott, C.; Wilson, S.H.

    1987-02-10

    A new polyclonal antibody against the ..cap alpha..-polymerase catalytic polypeptide was prepared by using homogeneous HeLa cell..cap alpha..-polymerase. The antibody neutralized ..cap alpha..-polymerase activity and was strong and specific for the ..cap alpha..-polymerase catalytic polypeptide (M/sub r/ 183,000) in Western blot analysis of crude extracts of HeLa cells. The antibody was used to screen a cDNA library of newborn rat brain poly(A+) RNA in lambdagt11. A positive phage was identified and plaque purified. This phage, designated lambdapol..cap alpha..1.2, also was found to be positive with an antibody against Drosophila ..cap alpha..-polymerase. The insert in lambdapol..cap alpha..1.2 (1183 base pairs) contained a poly(A) sequence at the 3' terminus and a short in-phase open reading frame at the 5' terminus. A synthetic oligopeptide (eight amino acids) corresponding to the open reading frame was used to raise antiserum in rabbits. Antibody affinity purified from this serum was found to be immunoreactive against purified ..cap alpha..-polymerase by enzyme-linked immunosorbent assay and was capable of immunoprecipitating ..cap alpha..-polymerase. This indicated the lambdapol..cap alpha..1.2 insert encoded an ..cap alpha..-polymerase epitope and suggested that the cDNA corresponded to an ..cap alpha..-polymerase mRNA. This was confirmed in hybrid selection experiments using pUC9 containing the cDNA insert and poly(A+) RNA from newborn rat brain; the insert hybridized to mRNA capable of encoding ..cap alpha..-polymerase catalytic polypeptides. Northern blot analysis of rat brain poly(A+) RNA revealed that this mRNA is approx.5.4 kilobases.

  1. Detection of PrP**Sc in formalin-fixed tissues by western blot

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Formalin fixation is the most prevalent form of tissue preservative. As such, formalin fixed tissue represents an important source of archival material for study. Formalin fixation requires little environmental control and preserves the cellular architecture of a wide range of tissues, an importan...

  2. Detection of Traces of Ovalbumin and Casein in White and Red Wines by Quantitative Western Blotting.

    PubMed

    Meyer, Philippe; Zanetti, Simone

    2015-09-01

    Fining of wine with agents containing cow's milk or hen's egg white is a common and traditional procedure. In light of increasing food allergies all over the world, the presence of fining residues has been subject of intense debate. Switzerland does not make exception, and since 2009 the Federal Department of Home Affairs has modified its food regulations stating that the labels must show if traces of fining agents are present. Nevertheless, the application of this regulation is not based on an official analytical method. In this study we show that immunoblotting is an efficient technique to detect and quantify ovalbumin and casein residues in bottled wine. We showed that final filtration is an essential step to remove finings in red wine, and that overfining of white wine may result in fining residues in finished products. Finally, for the first time in Switzerland, 22 samples were taken by food safety inspectors and officially analyzed for the regional food control authority of the Canton of Vaud. These samples were allergen free, but a larger study is currently planned in collaboration with other regional authorities of Switzerland to complete these results and make a complete picture of the Swiss wine production.

  3. Western blotting of high and low molecular weight proteins using heat.

    PubMed

    Kurien, Biji T; Scofield, R Hal

    2015-01-01

    A method for the electrophoretic transfer of high and low molecular weight proteins to nitrocellulose membranes following sodium dodecyl sulfate (SDS) polyacrylamide gel is described here. The transfer was performed with heated (70-75 °C) normal transfer buffer from which methanol had been omitted. Complete transfer of high and low molecular weight antigens (molecular weight protein standards, a purified protein, and proteins from a human tissue extract) could be carried out in 10 min for a 7 % (0.75 mm) SDS polyacrylamide gel. For 10 and 12.5 % gels (0.75 mm) the corresponding time was 15 min. A complete transfer could be carried out in 20 min for 7, 10, and 12.5 % gels (1.5 mm gels). The permeability of the gel is increased by heat, such that the proteins trapped in the polyacrylamide gel matrix can be easily transferred to the membrane. The heat mediated transfer method was compared with a conventional transfer protocol, under similar conditions. The conventional method transferred minimal low molecular weight proteins while retaining most of the high molecular weight proteins in the gel. In summary, this procedure is particularly useful for the transfer of high molecular weight proteins, very rapid, and avoids the use of methanol.

  4. Mucin Agarose Gel Electrophoresis: Western Blotting for High-molecular-weight Glycoproteins.

    PubMed

    Ramsey, Kathryn A; Rushton, Zachary L; Ehre, Camille

    2016-06-14

    Mucins, the heavily-glycosylated proteins lining mucosal surfaces, have evolved as a key component of innate defense by protecting the epithelium against invading pathogens. The main role of these macromolecules is to facilitate particle trapping and clearance while promoting lubrication of the mucosa. During protein synthesis, mucins undergo intense O-glycosylation and multimerization, which dramatically increase the mass and size of these molecules. These post-translational modifications are critical for the viscoelastic properties of mucus. As a result of the complex biochemical and biophysical nature of these molecules, working with mucins provides many challenges that cannot be overcome by conventional protein analysis methods. For instance, their high-molecular-weight prevents electrophoretic migration via regular polyacrylamide gels and their sticky nature causes adhesion to experimental tubing. However, investigating the role of mucins in health (e.g., maintaining mucosal integrity) and disease (e.g., hyperconcentration, mucostasis, cancer) has recently gained interest and mucins are being investigated as a therapeutic target. A better understanding of the production and function of mucin macromolecules may lead to novel pharmaceutical approaches, e.g., inhibitors of mucin granule exocytosis and/or mucolytic agents. Therefore, consistent and reliable protocols to investigate mucin biology are critical for scientific advancement. Here, we describe conventional methods to separate mucin macromolecules by electrophoresis using an agarose gel, transfer protein into nitrocellulose membrane, and detect signal with mucin-specific antibodies as well as infrared fluorescent gel reader. These techniques are widely applicable to determine mucin quantitation, multimerization and to test the effects of pharmacological compounds on mucins.

  5. Analysis of casein using two-dimensional electrophoresis, western blot, and computer imaging.

    PubMed

    Goldfarb, M F

    2001-01-01

    Casein from milk of 20 mothers, 8 weeks postpartum, was analyzed using 2-dimensional electrophoresis. The casein micelle under 2-D denaturing conditions gives approximately 15 major spots. Beta-Casein, kappa-casein, para-kappa-casein, and casein peptides can be tentatively identified by molecular size, isoelectric point, and spot shape characteristics. All spots are not present in all samples. Two spots thought to represent different phosphorylated forms of beta-casein were selected for area and density measurements by means of the NIH-Image program. The larger area at approximately 26 kDa, isoelectric point 5.3, consisted of 1 to 3 closely linked spots, perhaps reflecting phenotype. The smaller spot 24kDa, isoelectric point 5.4, was always a single unit. Visual scanning of gels suggests a wide range of casein concentration. Computer comparison will allow a statistical analysis.

  6. Active basement uplift as seen with cosmogenic lenses: the Sierra Pie de Palo case (Western Argentina)

    NASA Astrophysics Data System (ADS)

    Siame, L. L.; Sébrier, M.; Costa, C. H.; Ahumada, E. A.; Bellier, O.

    2013-12-01

    The Andean foreland of western Argentina (28°S-33°S) corresponds to retroarc deformations associated with the ongoing flat subduction of the Nazca plate beneath the South American lithosphere, and associated with high levels of seismic activity and crustal active faulting. To improve earthquake source identification and characterization in the San Juan region, data from seismology, structural geology and quantitative geomorphology can be integrated and combined to provide a seismotectonic model. In this model, the Andean back-arc of western Argentina has to be regarded as an obliquely converging foreland where Plio-Quaternary deformations are partitioned between strike-slip and thrust motions that are localized on the E-verging, thin-skinned Argentine Precordillera, and the W-verging thick-skinned Sierras Pampeanas, respectively. In this domain, the Sierra Pie de Palo is a key structure playing a major role in the partitioning of the Plio-Quaternary deformations. Located in the westernmost Sierras Pampeanas, the Sierra Pie de Palo forms a NNE striking, 80 km-long and 35-40 km-wide, ellipsoid range that reaches elevation as high as 3162 m. This mountain range is an actively growing basement fold associated with a high level of seismic activity (e.g., the November 23, 1977, Caucete, Mw 7.4 earthquake). To evaluate the degree of tectonic activity around the Sierra Pie de Palo, we combined a detailed morphometric analysis of the topography together with in situ-produced cosmogenic 10Be concentrations measured in (1) bedrock outcrops corresponding to the exhumed erosional regional surface, (2) surface boulders abandoned on alluvial fans deformed by active faults, and (3) in fluvial sediments sampled at the outlets of selected watersheds that drains out from the Sierra Pie de Palo. All together, our results allows: (1) assessing quantitative constraints on the rate of tectonic and denudation processes that are responsible for the active growth and erosion of the Sierra

  7. Documenting Western Burrowing Owl Reproduction and Activity Patterns Using Motion-Activated Cameras

    SciTech Connect

    Hall, Derek B.; Greger, Paul D.

    2014-08-01

    We used motion-activated cameras to monitor the reproduction and patterns of activity of the Burrowing Owl (Athene cunicularia) above ground at 45 burrows in south-central Nevada during the breeding seasons of 1999, 2000, 2001, and 2005. The 37 broods, encompassing 180 young, raised over the four years represented an average of 4.9 young per successful breeding pair. Young and adult owls were detected at the burrow entrance at all times of the day and night, but adults were detected more frequently during afternoon/early evening than were young. Motion-activated cameras require less effort to implement than other techniques. Limitations include photographing only a small percentage of owl activity at the burrow; not detecting the actual number of eggs, young, or number fledged; and not being able to track individual owls over time. Further work is also necessary to compare the accuracy of productivity estimates generated from motion-activated cameras with other techniques.

  8. The Central-Western Mediterranean: Anomalous igneous activity in an anomalous collisional tectonic setting

    NASA Astrophysics Data System (ADS)

    Lustrino, Michele; Duggen, Svend; Rosenberg, Claudio L.

    2011-01-01

    The central-western Mediterranean area is a key region for understanding the complex interaction between igneous activity and tectonics. In this review, the specific geochemical character of several 'subduction-related' Cenozoic igneous provinces are described with a view to identifying the processes responsible for the modifications of their sources. Different petrogenetic models are reviewed in the light of competing geological and geodynamic scenarios proposed in the literature. Plutonic rocks occur almost exclusively in the Eocene-Oligocene Periadriatic Province of the Alps while relatively minor plutonic bodies (mostly Miocene in age) crop out in N Morocco, S Spain and N Algeria. Igneous activity is otherwise confined to lava flows and dykes accompanied by relatively greater volumes of pyroclastic (often ignimbritic) products. Overall, the igneous activity spanned a wide temporal range, from middle Eocene (such as the Periadriatic Province) to the present (as in the Neapolitan of southern Italy). The magmatic products are mostly SiO 2-oversaturated, showing calcalkaline to high-K calcalcaline affinity, except in some areas (as in peninsular Italy) where potassic to ultrapotassic compositions prevail. The ultrapotassic magmas (which include leucitites to leucite-phonolites) are dominantly SiO 2-undersaturated, although rare, SiO 2-saturated (i.e., leucite-free lamproites) appear over much of this region, examples being in the Betics (southeast Spain), the northwest Alps, northeast Corsica (France), Tuscany (northwest Italy), southeast Tyrrhenian Sea (Cornacya Seamount) and possibly in the Tell region (northeast Algeria). Excepted for the Alpine case, subduction-related igneous activity is strictly linked to the formation of the Mediterranean Sea. This Sea, at least in its central and western sectors, is made up of several young (< 30 Ma) V-shaped back-arc basins plus several dispersed continental fragments, originally in crustal continuity with the European

  9. Increasing western US forest wildfire activity: sensitivity to changes in the timing of spring.

    PubMed

    Westerling, Anthony LeRoy

    2016-06-01

    Prior work shows western US forest wildfire activity increased abruptly in the mid-1980s. Large forest wildfires and areas burned in them have continued to increase over recent decades, with most of the increase in lightning-ignited fires. Northern US Rockies forests dominated early increases in wildfire activity, and still contributed 50% of the increase in large fires over the last decade. However, the percentage growth in wildfire activity in Pacific northwestern and southwestern US forests has rapidly increased over the last two decades. Wildfire numbers and burned area are also increasing in non-forest vegetation types. Wildfire activity appears strongly associated with warming and earlier spring snowmelt. Analysis of the drivers of forest wildfire sensitivity to changes in the timing of spring demonstrates that forests at elevations where the historical mean snow-free season ranged between two and four months, with relatively high cumulative warm-season actual evapotranspiration, have been most affected. Increases in large wildfires associated with earlier spring snowmelt scale exponentially with changes in moisture deficit, and moisture deficit changes can explain most of the spatial variability in forest wildfire regime response to the timing of spring.This article is part of the themed issue 'The interaction of fire and mankind'.

  10. Increasing western US forest wildfire activity: sensitivity to changes in the timing of spring.

    PubMed

    Westerling, Anthony LeRoy

    2016-06-01

    Prior work shows western US forest wildfire activity increased abruptly in the mid-1980s. Large forest wildfires and areas burned in them have continued to increase over recent decades, with most of the increase in lightning-ignited fires. Northern US Rockies forests dominated early increases in wildfire activity, and still contributed 50% of the increase in large fires over the last decade. However, the percentage growth in wildfire activity in Pacific northwestern and southwestern US forests has rapidly increased over the last two decades. Wildfire numbers and burned area are also increasing in non-forest vegetation types. Wildfire activity appears strongly associated with warming and earlier spring snowmelt. Analysis of the drivers of forest wildfire sensitivity to changes in the timing of spring demonstrates that forests at elevations where the historical mean snow-free season ranged between two and four months, with relatively high cumulative warm-season actual evapotranspiration, have been most affected. Increases in large wildfires associated with earlier spring snowmelt scale exponentially with changes in moisture deficit, and moisture deficit changes can explain most of the spatial variability in forest wildfire regime response to the timing of spring.This article is part of the themed issue 'The interaction of fire and mankind'. PMID:27216510

  11. EarthScope Transportable Array Siting Outreach Activities in Alaska and Western Canada

    NASA Astrophysics Data System (ADS)

    Gardine, L.; Dorr, P. M.; Tape, C.; McQuillan, P.; Taber, J.; West, M. E.; Busby, R. W.

    2014-12-01

    The EarthScopeTransportable Array is working to locate over 260 stations in Alaska and western Canada. In this region, new tactics and partnerships are needed to increase outreach exposure. IRIS and EarthScope are partnering with the Alaska Earthquake Center, part of University of Alaska Geophysical Institute, to spread awareness of Alaska earthquakes and the benefits of the Transportable Array for Alaskans. Nearly all parts of Alaska are tectonically active. The tectonic and seismic variability of Alaska requires focused attention at the regional level, and the remoteness and inaccessibility of most Alaska villages and towns often makes frequent visits difficult. For this reason, Alaska outreach most often occurs at community events. When a community is accessible, every opportunity to engage the residents is made. Booths at state fairs and large cultural gatherings, such as the annual convention of the Alaska Federation of Natives, are excellent venues to distribute earthquake information and to demonstrate a wide variety of educational products and web-based applications related to seismology and the Transportable Array that residents can use in their own communities. Region-specific publications have been developed to tie in a sense of place for residents of Alaska. The Alaska content for IRIS's Active Earth Monitor will emphasize the widespread tectonic and seismic features and offer not just Alaska residents, but anyone interested in Alaska, a glimpse into what is going on beneath their feet. The concerted efforts of the outreach team will have lasting effects on Alaskan understanding of the seismic hazard and tectonics of the region. Efforts to publicize the presence of the Transportable Array in Alaska, western Canada, and the Lower 48 also continue. There have been recent articles published in university, local and regional newspapers; stories appearing in national and international print and broadcast media; and documentaries produced by some of the world

  12. Active normal faulting along the Mt. Morrone south-western slopes (central Apennines, Italy)

    NASA Astrophysics Data System (ADS)

    Gori, Stefano; Giaccio, Biagio; Galadini, Fabrizio; Falcucci, Emanuela; Messina, Paolo; Sposato, Andrea; Dramis, Francesco

    2011-01-01

    In the present work we analyse one of the active normal faults affecting the central Apennines, i.e. the Mt. Morrone normal fault system. This tectonic structure, which comprises two parallel, NW-SE trending fault segments, is considered as potentially responsible for earthquakes of magnitude ≥ 6.5 and its last activation probably occurred during the second century AD. Structural observations performed along the fault planes have allowed to define the mainly normal kinematics of the tectonic structure, fitting an approximately N 20° trending extensional deformation. Geological and geomorphological investigations performed along the whole Mt. Morrone south-western slopes permitted us to identify the displacement of alluvial fans, attributed to Middle and Late Pleistocene by means of tephro-stratigraphic analyses and geomorphological correlations with dated lacustrine sequences, along the western fault branch. This allowed to evaluate in 0.4 ± 0.07 mm/year the slip rate of this segment. On the other hand, the lack of synchronous landforms and/or deposits that can be correlated across the eastern fault segment prevented the definition of the slip rate related to this fault branch. Nevertheless, basing on a critical review of the available literature dealing with normal fault systems evolution, we hypothesised a total slip rate of the fault system in the range of 0.4 ± 0.07 to 0.8 ± 0.09 mm/year. Moreover, basing on the length at surface of the Mt. Morrone fault system (i.e. 22-23 km) we estimated the maximum expected magnitude of an earthquake that might originate along this tectonic structure in the order of 6.6-6.7.

  13. Evaluation of antiviral activity of aqueous extracts from Achyrocline satureioides against Western equine encephalitis virus.

    PubMed

    Sabini, María Carola; Escobar, Franco Matías; Tonn, Carlos Eugenio; Zanon, Silvia Matilde; Contigiani, Marta Silvia; Sabini, Liliana Inés

    2012-01-01

    Achyrocline satureioides (Asteraceae) is a medicinal plant traditionally used in Argentina for the treatment of intestinal infections and various digestive disorders. Its infusion is widely utilised for respiratory problems and viral infections. The objective of this study was to investigate cytotoxicity, virucidal and antiviral properties of the cold aqueous extract (CAE) and hot aqueous extract (HAE) of this plant against Western equine encephalitis virus (WEEV). Cytotoxicity in Vero cells was evaluated by maximum non-cytotoxic concentration (MNCC), neutral red (NR) uptake and MTT reduction methods. To study the antiviral activity of aqueous extracts, plaque reduction assay was performed after pre-treatment of host cells, adsorption, penetration and post-penetration of the virus. Extracellular virus inactivation was also analysed by the same method. Extracts showed strong inhibitory activity after virus penetration with selective index values of 32 (NR) and 63.3 (MTT) for the CAE, and 16.2 (NR) and 24.3 (MTT) for the HAE. Both extracts exhibited virucidal action with lower efficacy than their antiviral properties. The present results demonstrate that aqueous extracts of A. satureioides are active against WEEV. Further studies are needed in order to identify which compounds could be responsible for this effect, and how they exert antiviral action.

  14. Discovery of Active Hydrothermal Sites Along the Mariana Volcanic Arc, Western Pacific Ocean

    NASA Astrophysics Data System (ADS)

    Baker, E. T.; Embley, R. W.; Resing, J. A.; Lupton, J. E.; Massoth, G. J.; de Ronde, C. E.; Nakamura, K.; Walker, S. L.

    2003-12-01

    Some 20,000 km of volcanic arcs, roughly one-third the total length of the global midocean ridge (MOR) system, rim the western Pacific Ocean. But compared to 25 years of hydrothermal investigations along MORs, exploration of similar activity on the estimated 600 submarine arc volcanoes is only beginning. In February 2003, as part of the Submarine Ring of Fire project funded by NOAA's Ocean Exploration Program, we made the first systematic survey of hydrothermal activity along the 1270-km-long Mariana intraoceanic volcanic arc, which lies almost entirely within the US EEZ. Prior fieldwork had documented active (but low-temperature) hydrothermal discharge on only three volcanoes: Kasuga 2, Kasuga 3, and Esmeralda Bank. During the cruise, we conducted 70 CTD operations over more than 50 individual volcanoes from 13° N to 23° N, plus a continuous CTD survey along 75 km of the back-arc spreading center (13° 15'N to 13° 41'N) adjacent to the southern end of the arc. We found evidence for active hydrothermal venting at 11 submarine volcanoes with summit (or caldera floor) depths ranging from 50 to 1550 m. Two additional sites were identified on the back-arc spreading center. Ongoing analyses of collected water samples could increase these totals. Our results confirmed continuing hydrothermal activity at Kasuga 2 (but not Kasuga 3) and Esmeralda Bank, in addition to newly discovered sites on nine other volcanoes. Many of these sites produce intense and widely dispersed plumes indicative of vigorous, high-temperature discharge. The volcanoes with active hydrothermal systems are about equally divided between those with and without summit calderas. The addition of the Marianas data greatly improves our view of hydrothermal sources along arcs. The 20,000 km of Pacific arcs can be divided between 6380 km of intraoceanic (i.e., mostly submarine) arcs and 13,880 km of island (i.e., mostly subaerial) arcs. At present, ˜15% of the total length of Pacific arcs has been surveyed

  15. Low Prevalence of Ocular Chlamydia trachomatis Infection and Active Trachoma in the Western Division of Fiji

    PubMed Central

    Mudaliar, Umesh; Natutusau, Kinisimere; Pavluck, Alexandre L.; Willis, Rebecca; Alexander, Neal; Mabey, David C. W.; Cikamatana, Luisa; Kama, Mike; Rafai, Eric; Roberts, Chrissy H.; Solomon, Anthony W.

    2016-01-01

    Background Trachoma is the leading infectious cause of blindness and is caused by ocular infection with the bacterium Chlamydia trachomatis (Ct). While the majority of the global disease burden is found in sub-Saharan Africa, the Western Pacific Region has been identified as trachoma endemic. Population surveys carried out throughout Fiji have shown an abundance of both clinically active trachoma and trachomatous trichiasis in all divisions. This finding is at odds with the clinical experience of local healthcare workers who do not consider trachoma to be highly prevalent. We aimed to determine whether conjunctival infection with Ct could be detected in one administrative division of Fiji. Methods A population-based survey of 2306 individuals was conducted using the Global Trachoma Mapping Project methodology. Population prevalence of active trachoma in children and trichiasis in adults was estimated using the World Health Organization simplified grading system. Conjunctival swabs were collected from 1009 children aged 1–9 years. DNA from swabs was tested for the presence of the Ct plasmid and human endogenous control. Results The prevalence of active trachoma in 1–9 year olds was 3.4%. The age-adjusted prevalence was 2.8% (95% CI: 1.4–4.3%). The unadjusted prevalence of ocular Ct infection in 1–9 year-olds was 1.9% (19/1009), and the age-adjusted infection prevalence was 2.3% (95% CI: 0.4–2.5%). The median DNA load was 41 Ct plasmid copies per swab (min 20, first quartile 32, mean 6665, third quartile 161, max 86354). There was no association between current infection and follicular trachoma. No cases of trachomatous trichiasis were identified. Discussion The Western Division of Fiji has a low prevalence of clinical trachoma. Ocular Ct infections were observed, but they were predominantly low load infections and were not correlated with clinical signs. Our study data suggest that trachoma does not meet the WHO definition of a public health problem in

  16. Measles Elimination Activities in the Western Pacific Region: Experience from the Republic of Korea

    PubMed Central

    Choe, Young June; Jee, Youngmee; Oh, Myoung-don

    2015-01-01

    We describe the global status of measles control and elimination, including surveillance and vaccination coverage data provided by the World Health Organization (WHO). Since 2000, two doses of measles vaccine (MCV2) became recommended globally and the achievement of high vaccination coverage has led to dramatic decrease in the measles incidence. Our finding indicates that, in the Western Pacific Region (WPR), substantial progress has been made to control measles transmission in some countries; however, the measles virus continues to circulate, causing outbreaks. The Republic of Korea (ROK) experienced a series of resurgence of measles due to the importation and healthcare-associated transmission in infants, however overall incidence and surveillance indicators met the WHO criteria for measles elimination. The ROK was verified to be measles-free along with Australia, Mongolia, and Macau, China in 2014. One of the effective elimination activities was the establishment of solid keep-up vaccination system in school settings. The lessons learnt from the measles elimination activities in Korea may contribute to enhancing the surveillance schemes and strengthening of vaccination programs in member countries and areas of WPR. PMID:26617443

  17. Decadal changes in tropical cyclone activity over the western North Pacific in the late 1990s

    NASA Astrophysics Data System (ADS)

    He, Haozhe; Yang, Jing; Gong, Daoyi; Mao, Rui; Wang, Yuqing; Gao, Miaoni

    2015-12-01

    A pronounced decadal change in tropical cyclone (TC) activity over the western North Pacific (WNP) in the late 1990s was identified. Based on a comparison of the two epochs that occurred before and after the late 1990s, the TC genesis number exhibited an evident decrease over the southern WNP (S-WNP: 5°-20°N, 105°-170°E) and an increase over the northern WNP (N-WNP: 20°-25°N, 115°-155°E), which partly corresponded to a significant northward migration in the seasonal mean latitudinal location of TC genesis, i.e., from 17.2°N to 18.7°N. After the late 1990s, the northwestward-moving track became the most dominant track mode, accompanied by the weakening of both the westward-moving track and the northeastward-recurving track. Meanwhile, the TC occurrence frequency (TCF) experienced evident increases over southeastern China and the Okinawa islands, while prominent decreases occurred over the South China Sea, the Philippine Sea, Japan and east of Japan. Changes in the TCF were determined by TC genesis changes, TC track shifts and variations in regional TC durations, which were all ascribed to the decadal change in tropical Indo-Pacific sea surface temperature. The full picture on the decadal changes in the WNP TC activity revealed in this study may provide useful guidance for regional TC seasonal forecasts and future projections.

  18. Measles Elimination Activities in the Western Pacific Region: Experience from the Republic of Korea.

    PubMed

    Choe, Young June; Jee, Youngmee; Oh, Myoung-don; Lee, Jong-Koo

    2015-11-01

    We describe the global status of measles control and elimination, including surveillance and vaccination coverage data provided by the World Health Organization (WHO). Since 2000, two doses of measles vaccine (MCV2) became recommended globally and the achievement of high vaccination coverage has led to dramatic decrease in the measles incidence. Our finding indicates that, in the Western Pacific Region (WPR), substantial progress has been made to control measles transmission in some countries; however, the measles virus continues to circulate, causing outbreaks. The Republic of Korea (ROK) experienced a series of resurgence of measles due to the importation and healthcare-associated transmission in infants, however overall incidence and surveillance indicators met the WHO criteria for measles elimination. The ROK was verified to be measles-free along with Australia, Mongolia, and Macau, China in 2014. One of the effective elimination activities was the establishment of solid keep-up vaccination system in school settings. The lessons learnt from the measles elimination activities in Korea may contribute to enhancing the surveillance schemes and strengthening of vaccination programs in member countries and areas of WPR.

  19. Effects of ammonium and nitrate on nutrient uptake and activity of nitrogen assimilating enzymes in western hemlock

    SciTech Connect

    Knoepp, J.D.; Turner, D.P.; Tingey, D.T.

    1993-01-01

    Western hemlock seedlings were grown in nutrient solutions with ammonium, nitrate or ammonium plus nitrate as nitrogen sources. The objectives were to examine (1) possible selectivity for ammonium or nitrate as an N source, (2) the maintenance of charge balance during ammonium and nitrate uptake, and (3) the activity of the nitrogen assimilating enzymes, nitrate reductase, glutamine synthetase, and glutamine dehydrogenase, in relation to the uptake of different nitrogen sources. The uptake studies revealed that western hemlock takes up ammonium faster than nitrate and that ammonium partially inhibits nitrate uptake. Nitrate reductase activity varied with nitrate availability in root tissue, but showed no response in needles, indicating that most nitrate is reduced in the roots. Results indicate that western hemlock may be adapted to sites where NH(4+) is the predominate N source.

  20. Acceleration of thaw slump activity in glaciated landscapes of the Western Canadian Arctic

    NASA Astrophysics Data System (ADS)

    Segal, Rebecca A.; Lantz, Trevor C.; Kokelj, Steven V.

    2016-03-01

    Climate change is increasing the frequency and intensity of thermokarst, but the influences of regional climate and physiography remain poorly understood. Retrogressive thaw slumping is one of the most dynamic forms of thermokarst and affects many areas of glaciated terrain across northwestern Canada. In this study, we used airphotos and satellite imagery to investigate the influence of climate and landscape factors on thaw slump dynamics. We assessed slump size, density, and growth rates in four regions of ice-rich terrain with contrasting climate and physiographic conditions: the Jesse Moraine, the Tuktoyaktuk Coastlands, the Bluenose Moraine, and the Peel Plateau. Observed increases in: (1) the area impacted by slumps (+2 to +407%), (2) average slump sizes (+0.31 to +1.82 ha), and (3) slump growth rates (+169 to +465 m2 yr-1) showed that thermokarst activity is rapidly accelerating in ice-rich morainal landscapes in the western Canadian Arctic, where slumping has become a dominant driver of geomorphic change. Differences in slump characteristics among regions indicate that slump development is strongly influenced by topography, ground ice conditions, and Quaternary history. Observed increases in slump activity occurred in conjunction with increases in air temperature and precipitation, but variation in slump activity among the four regions suggests that increased precipitation has been an important driver of change. Our observation that the most rapid intensification of slump activity occurred in the coldest environment (the Jesse Moraine on Banks Island) indicates that ice-cored landscapes in cold permafrost environments are highly vulnerable to climate change.

  1. On Active Layer Environments and Processes in Western Dronning Maud Land, Antarctica

    NASA Astrophysics Data System (ADS)

    Hansen, C. D.; Meiklejohn, I.; Nel, W.

    2012-12-01

    The current understanding of Antarctic permafrost is poor, particularly regarding its evolution, the current thermal characteristics, and relationships with pedogenesis, hydrology, geomorphic, dynamics, biotic activity and response to global changes. Results from borehole temperature measurements over a four-year period in Western Dronning Maud Land suggest that the active layer depth is dependent on the substrate, latitude, altitude and the volume of ground exposed; the latter alludes to the potential impact of surrounding ice on the ground thermal regime. The active layer depths at the monitoring sites, varied between 16 cm at Vesleskarvet, a small nunatak at 850 masl to 28 cm in granitic till at Jutulsessen (1 270 masl). The mean near surface (1.5 cm depth) ground temperatures from 2009 to 2012 in the region have a narrow range from -16.4°C at 850m to -17.5°C at 1270 masl. Permafrost temperatures for the same locations vary between -16.3°C and -18.3°C. While little variability exists between the mean temperatures at the study locations, each site is distinct and seasonal and shorter-term frost cycles have produced landforms that are characteristic of both permafrost and diurnal frost environments. One of the key aspects of investigation is the control that the active layer has on autochthonous blockfield development in the region. The, thus far, exploratory research is being used to understand controls on the landscape and the relationship between distribution and abundance of biota. Given the rapidly changing climates in the region, improving knowledge of what drives patterns of biodiversity at a local and regional scale is vital to assess consequences of environmental change.

  2. 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

  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. Primary centers and secondary concentrations of tectonic activity through time in the western hemisphere of Mars

    USGS Publications Warehouse

    Anderson, R.C.; Dohm, J.M.; Golombek, M.P.; Haldemann, A.F.C.; Franklin, B.J.; Tanaka, K.L.; Lias, J.; Peer, B.

    2001-01-01

    Five main stages of radial and concentric structures formed around Tharsis from the Noachian through the Amazonian as determined by geologic mapping of 24,452 structures within the stratigraphic framework of Mars and by testing their radial and concentric orientations. Tectonic activity peaked in the Noachian (stage 1) around the largest center, Claritas, an elongate center extending more than 20?? in latitude and defined by about half of the total grabens which are concentrated in the Syria Planum, Thaumasia, and Tempe Terra regions. During the Late Noachian and Early Hesperian (stage 2), extensional structures formed along the length of present-day Valles Marineris and in Thaumasia (with a secondary concentration near Warrego Vallis) radial to a region just to the south of the central margin of Valles Marineris. Early Hesperian (stage 3) radial grabens in Pavonis, Syria, Ulysses, and Tempe Terra and somewhat concentric wrinkle ridges in Lunae and Solis Plana and in Thaumasia, Sirenum, Memnonia, and Amazonis are centered northwest of Syria with secondary centers at Thaumasia, Tempe Terra, Ulysses Fossae, and western Valles Marineris. Late Hesperian/Early Amazonian (stage 4) structures around Alba Patera, the northeast trending alignment of Tharsis Montes, and Olympus Mons appears centered on Alba Patera. Stage 5 structures (Middle-Late Amazonian) represent the last pulse of Tharsis-related activity and are found around the large shield volcanoes and are centered near Pavonis Mons. Tectonic activity around Tharsis began in the Noachian and generally decreased through geologic time to the Amazonian. Statistically significant radial distributions of structures formed during each stage, centered at different locations within the higher elevations of Tharsis. Secondary centers of radial structures during many of the stages appear related to previously identified local magmatic centers that formed at different times and locations throughout Tharsis. Copyright 2001 by

  5. An Evaluation of Activated Bismuth Isotopes in Environmental Samples From the Former Western Pacific Proving Grounds

    SciTech Connect

    Robison, W.L.; Brunk, J.A.; Jokela, T.A.

    2000-03-21

    {sup 207}Bi (t{sub 1/2}=32.2 y) was generated by activation of weapons material during a few ''clean'' nuclear tests at the U.S. Western Pacific Proving Grounds of Enewetak and Bikini Atolls. The radionuclides first appeared in the Enewetak environment during 1958 and in the environment of Bikini during 1956. Crater sediments from Bikini with high levels of {sup 207}Bi were analyzed by gamma spectrometry in an attempt to determine the relative concentrations of {sup 208}Bi (t{sup 1/2} = 3.68 x 10{sup 5} y). The bismuth isotopes were probably generated during the ''clean'', 9.3 Mt Poplar test held on 7/12/58. The atom ratio of {sup 208}Bi to {sup 207}Bi (R value) ranges from {approx}12 to over 200 in sections of core sediments from the largest nuclear crater at Bikini atoll. The presence of bismuth in the device is suggested to account for R values in excess of 10.

  6. Hotei Arcus And Western Xanadu, Titan: Evidence For Cryovolcanic Activity From Radar

    NASA Astrophysics Data System (ADS)

    Lopes, Rosaly M. C.; Wall, S. D.; Stofan, E.; Wood, C.; Radebaugh, J.; Mitchell, K. L.; Stiles, B.; Nelson, R. M.; Kamp, L.; Janssen, M.; Lorenz, R. D.; Lunine, J.; Farr, T.; Mitri, G.; Paillou, P.; Paganelli, F.; Cassini RADAR Team

    2008-09-01

    Several surface features interpreted as cryovolcanic in origin have been observed on Titan's surface by both Cassini's RADAR (in SAR mode) and VIMS instruments throughout the Cassini prime mission. The Titan flyby T41 of 22 February 2008 includes a SAR image of part of Hotei Arcus, a semi-circular albedo feature, some 650 km in length along the arc, centered at 26S 79W. A second SAR image of Hotei was acquired May 12, 2008 on flyby T43. These images show that the arcuate southern boundary of Hotei, also seen in ISS data, appears somewhat mountainous in the SAR imagery, and 5 distinct narrow channels, presumably fluvial, flow radially inwards. In the center of the arc, the images reveal lobate, flowlike features that embay surrounding terrains and the channels. Analysis of these features suggest that they are of cryovolcanic origin and younger than surrounding terrain. Their appearance is superficially similar to a region in western Xanadu at 10S 140W imaged by RADAR on flyby T13, on Apr 30, 2006. These two regions are morphologically unlike most of the other cryovolcanic regions so far seen on Titan. Both regions correspond to those identified by the Cassini VIMS as having anomalous and variable infrared brightness, probably due to recent cryovolcanic activity. The RADAR images provide morphological evidence that is consistent with cryovolcanism.

  7. Change of tropical cyclone activity by Pacific-Japan teleconnection pattern in the western North Pacific

    NASA Astrophysics Data System (ADS)

    Choi, Ki-Seon; Wu, Chun-Chieh; Cha, Eun-Jeong

    2010-10-01

    This study shows that the Pacific-Japan (PJ) teleconnection pattern has a significant influence on tropical cyclone (TC) activities over the western North Pacific (WNP) during the boreal summer (July, August, and September). During positive (negative) PJ phase, TCs form at a more northward (southward) location, recurve at a more northeastward (southwestward) location, and frequently pass over the northeast Asian (southeast Asian) region, including Korea and Japan (South China Sea and southern China). In particular, this difference in the TC track between the two phases is observed as a dipole-like pattern between the regions of Southeast and Northeast Asia. The TC characteristics during the positive PJ phase are caused by the following two stronger atmospheric circulations over the WNP: an anticyclonic circulation centered to the east of Japan and a cyclonic circulation centered to the east of Taiwan. The southeasterly between these two circulations serves as steering flow that TCs move northward toward Korea and Japan from the northeast of the Philippines. Conversely, TCs during the negative PJ phase mainly move westward toward the South China Sea and southern China by the easterly from a stronger anticyclonic circulation centered to the east of Taiwan. As a result of this feature of TC track during the negative PJ phase, TC lifetime is shorter and TC intensity is weaker.

  8. Possible relationship between western North Pacific tropical cyclone activity and Arctic Oscillation

    NASA Astrophysics Data System (ADS)

    Choi, Ki-Seon; Byun, Hi-Ryong

    2010-05-01

    This study found that Arctic Oscillation (AO) has a significant influence on tropical cyclone (TC) activities in the western North Pacific during the boreal summer (July, August, and September). During low- (high-) AO years, more TCs formed over east (west) of 150° E, recurved in the east (west), and passed over the midlatitudes (southeast Asian region), including Korea and Japan (South China Sea and south China), compared to the high- (low-) AO years. In particular, the TC passage frequency difference between the two periods showed a dipole-like pattern between the regions of Southeast and Northeast Asia. The differences between these two periods were caused by a stronger anticyclonic circulation located around Korea and Japan during high-AO years. This circulation played a significant role in blocking the movement of TCs toward Korea and Japan during high-AO years. Instead, TCs moved westward toward the SCS and southern China along the easterly and southeasterly steering flow of this anticyclonic circulation. As a result, TC lifetime and intensity were shorter and weaker during high-AO years.

  9. Seismicity on the western Greenland Ice Sheet: Surface fracture in the vicinity of active moulins

    SciTech Connect

    Carmichael, Joshua D.; Joughin, Ian; Behn, Mark D.; Das, Sarah; King, Matt A.; Stevens, Laura; Lizarralde, Dan

    2015-06-25

    We analyzed geophone and GPS measurements collected within the ablation zone of the western Greenland Ice Sheet during a ~35 day period of the 2011 melt season to study changes in ice deformation before, during, and after a supraglacial lake drainage event. During rapid lake drainage, ice flow speeds increased to ~400% of winter values, and icequake activity peaked. At times >7 days after drainage, this seismicity developed variability over both diurnal and longer periods (~10 days), while coincident ice speeds fell to ~150% of winter values and showed nightly peaks in spatial variability. Approximately 95% of all detected seismicity in the lake basin and its immediate vicinity was triggered by fracture propagation within near-surface ice (<330 m deep) that generated Rayleigh waves. Icequakes occurring before and during drainage frequently were collocated with the down flow (west) end of the primary hydrofracture through which the lake drained but shifted farther west and outside the lake basin after the drainage. We interpret these results to reveal vertical hydrofracture opening and local uplift during the drainage, followed by enhanced seismicity and ice flow on the downstream side of the lake basin. This region collocates with interferometric synthetic aperture radar-measured speedup in previous years and could reflect the migration path of the meltwater supplied to the bed by the lake. The diurnal seismic signal can be associated with nightly reductions in surface melt input that increase effective basal pressure and traction, thereby promoting elevated strain in the surficial ice.

  10. Assessment of relative tectonic activity in the Trichonis Lake graben (Western Greece) using geomorphometry

    NASA Astrophysics Data System (ADS)

    Karymbalis, Efthimios; Valkanou, Kanella; Fubelli, Giandomenico; Ferentinou, Maria; Giles, Philip; Papanastassiou, Dimitris; Gaki-Papanastassiou, Kalliopi; Tsanakas, Konstantinos

    2016-04-01

    In tectonically active areas fluvial systems and mountain fronts are controlled by the type, geometry, and recent activity of faults. The aim of this study is to investigate the contribution of neotectonics to the development of the fluvial landscape of the broader Trichonis Lake area (located in western continental Greece) through quantitative geomorphological analysis. The Trichonis Lake graben is a well-known tectonic depression of Quaternary age, which cuts across the early Tertiary NW-SE fold and thrust structures of the Pindos Mountain belt. It strikes WNW-ESE for a distance of 32 km and has a width of 10 km. The graben at the north and south flanks of the lake is bounded by E-W and NW-SE trending faults. Recent seismic activity (a shallow earthquake sequence in 1975 and a 2007 earthquake swarm) showed the existence of a NNW-SSE normal fault that dips to the NE and bounds the south-eastern shore of the lake. The studied catchments are developed on the hanging walls of these active normal faults. To evaluate the relative tectonic activity in the study area, various morphometric indices were measured for 35 catchments (slope of the valley sides of the catchment, hypsometric integral, catchment asymmetry factor, relief ratio, Melton's ruggedness number, stream-gradient index, ratio of valley floor width to valley height, and catchment shape) and 20 mountain fronts (mountain-front sinuosity index) around the lake. For the measurement of the geomorphometric variables a digital elevation model (DEM) with 2-m spatial resolution was derived from topographic maps at 1:5000 scale with 4-m contour lines, and a series of maps showing the spatial distribution of the variables were produced in a GIS environment. For each morphometric variable the catchments were classified into three classes. The combination of these morphometric variables allowed us to yield two new indices of relative tectonic activity (named IRTA - Index of Relative Tectonic Activity and IAT - Index of

  11. Possible effects of ongoing and predicted climate change on snow avalanche activity in western Norway

    NASA Astrophysics Data System (ADS)

    Laute, Katja; Beylich, Achim A.

    2016-04-01

    As snow avalanche formation is mainly governed by meteorological conditions as, e.g., air temperature fluctuations, heavy precipitation and wind conditions, it is likely that the frequency and magnitude of both ordinary and extreme snow avalanche events is modified through the documented effects of current and future climate change. In the Northern Hemisphere, 1983-2013 was likely the warmest 30-year period of the last 1400 years (IPCC, 2013). Meteorological records of western Norway show the general trend that the last 100 years, especially the last three decades, have been warmer and wetter than the time periods before. However, it is not evident that snow avalanche activity will increase in the near future. Today, the number of studies assessing the impact of climate change on the occurrence and magnitude of snow avalanches is limited. This work focuses on recent and possible future effects of climate change on snow avalanche activity along the western side of the Jostedalsbreen ice cap representing one of the areas with the highest snow avalanche activity in entire Norway. We have analyzed long-term homogenized meteorological data from five meteorological stations in different elevations above sea level, three of them with a long-term record of 120 years (1895-2015). In addition to the statistical analyses of long-term datasets, gained results and insights from a four-year (2009-2012) high-resolution snow avalanche monitoring study conducted in the same study area are incorporated. The statistical analyses of mean monthly air temperature, monthly precipitation sums and mean monthly snow depths showed that there is a trend of increasing air temperatures and precipitation sums whereas no clear trend was found for mean snow depths. Magnitude-frequency analyses conducted for three defined time intervals (120, 90, 60 years) of monthly precipitation sums exhibit an increase of precipitation especially during the last 30 years with the tendency that more precipitation

  12. Tectonic and Structural Controls of Geothermal Activity in the Great Basin Region, Western USA

    NASA Astrophysics Data System (ADS)

    Faulds, J. E.; Hinz, N.; Kreemer, C. W.

    2012-12-01

    We are conducting a thorough inventory of structural settings of geothermal systems (>400 total) in the extensional to transtensional Great Basin region of the western USA. Most of the geothermal systems in this region are not related to upper crustal magmatism and thus regional tectonic and local structural controls are the most critical factors controlling the locations of the geothermal activity. A system of NW-striking dextral faults known as the Walker Lane accommodates ~20% of the North American-Pacific plate motion in the western Great Basin and is intimately linked to N- to NNE-striking normal fault systems throughout the region. Overall, geothermal systems are concentrated in areas with the highest strain rates within or proximal to the eastern and western margins of the Great Basin, with the high temperature systems clustering in transtensional areas of highest strain rate in the northwestern Great Basin. Enhanced extension in the northwestern Great Basin probably results from the northwestward termination of the Walker Lane and the concomitant transfer of dextral shear into west-northwest directed extension, thus producing a broad transtensional region. The capacity of geothermal power plants also correlates with strain rates, with the largest (hundreds of megawatts) along the Walker Lane or San Andreas fault system, where strain rates range from 10-100 nanostrain/yr to 1,000 nanostrain/yr, respectively. Lesser systems (tens of megawatts) reside in the Basin and Range (outside the Walker Lane), where local strain rates are typically < 10 nanostrain/yr. Of the 250+ geothermal fields catalogued, step-overs or relay ramps in normal fault zones serve as the most favorable setting, hosting ~32% of the systems. Such areas have multiple, overlapping fault strands, increased fracture density, and thus enhanced permeability. Other common settings include a) intersections between normal faults and strike-slip or oblique-slip faults (27%), where multiple minor

  13. Alkaline phosphatase activity in the western English Channel: Elevations induced by high summertime rainfall

    NASA Astrophysics Data System (ADS)

    Rees, Andrew P.; Hope, Sam B.; Widdicombe, Claire E.; Dixon, Joanna L.; Woodward, E. Malcolm S.; Fitzsimons, Mark F.

    2009-03-01

    Alkaline phosphatase activity (APA) was determined in bulk particulate material and in a single-cell (ELF) assay at station L4 in the western English Channel during the summer of 2007. Throughout this period, the UK experienced its heaviest summertime rainfall since records began in 1914; with the result that riverine run-off into coastal waters was also elevated relative to long-term averages. Between May and August 2007, three distinct periods of elevated river run-off were observed which resulted in salinity minima at L4 on days 141, 190 and 232. An extended period of high river run-off between days 170 and 210 was responsible for decreases in near-surface salinity at L4 from 35.2068 to a minimum on day 190 of 34.7422. This contributed to the development of haline stratification which supported the development of an intense bloom of the centric diatom Chaetoceros debelis, with maximum observed chlorophyll a concentration of 8.69 μg l -1. Minima in salinity, and maxima in chlorophyll concentration on day 190 were coincident with a peak in river-derived dissolved inorganic nitrogen (DIN) of 1.9 μmol l -1 which was >5 times greater than the summertime mean and 24 times the concentrations experienced at L4 on weeks immediately before and after. There was no accompanying increase in dissolved inorganic phosphorus (DIP), and the DIN:DIP ratio increased to 49. With the inherent phosphorus stress that this caused, rates of APA increased from <4 to 42.4 nmolP l -1 h -1. ELF analysis on day 197 identified two taxa actively expressing alkaline phosphatase: the dinoflagellate Prorocentrum micans and ciliate Tiarana sp.

  14. 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

  15. Geologic Mapping to Constrain the Sources and Timing of Fluvial Activity in Western Ladon Basin, Mars

    NASA Astrophysics Data System (ADS)

    Weitz, C. M.; Wilson, S. A.; Irwin, R. P.; Grant, J. A.

    2016-06-01

    We are mapping two quadrangles in Margaritifer Terra (-15032 and -20032) to define the evolution of the western Ladon basin region as it relates to fluvial/alluvial events occurring on surrounding surfaces.

  16. Soil microbial activity and structure in mineralized terranes of the Western US

    NASA Astrophysics Data System (ADS)

    Blecker, S. W.; Stillings, L. L.; Decrappeo, N.; Ippolito, J.

    2009-12-01

    Mineralized terranes (areas enriched in metal-bearing minerals) occur throughout the Western US, and are characterized by highly variable soil trace metal concentrations across small spatial scales. Assuming that non-lithologic (extrinsic) soil forming factors are relatively constant between mineralized and unmineralized zones, these mineralized areas allowed us to evaluate the effect of lithology on soil microbial activity. We established the following study sites: 1) sage-grassland on a Mo/Cu deposit (Battle Mountain, NV); 2) pine-chaparral on Ni/Cr bearing rocks (Chinese Camp, CA); and 3) two pine woodland sites on acid-sulfate altered rocks (Reno, NV; Bridgeport, CA). Microbial, physical and chemical measurements were performed on soils from undisturbed mineralized areas and adjacent unmineralized areas to determine baseline conditions for comparison to sites disturbed by mining. A host of abiotic soil parameters, along with bioavailable (diethylenetriaminepentaacetic acid (DTPA)-extractable) and total metals, were measured to examine their correlation with the following measures of microbial activity: enzyme assays (arylsulfatase, phosphatase, fluorescein diacetate hydrolysis), C/N mineralization potential, C substrate utilization (Biolog Ecoplate), and microbial biomass and community structure (phospholipid fatty acid analysis). Within the Battle Mountain study area, both microbial activity and structure were statistically similar between mineralized and unmineralized soils. Nutrient and metal concentrations were also similar; the only differences being higher Cu and lower P in the mineralized soils. Within the Chinese Camp study area, soil organic carbon and total nitrogen concentrations were similar between the serpentine (Ni/Cr bearing) and adjacent andesite soils, while differences were noted for other nutrients (S, P, Ca, Mg). For the serpentine soils, Co, Fe, Mn, and Ni showed the strongest correlations with microbial activity, where Cr, Mn showed the

  17. Extraction of DNA from paraffin blocks for Southern blot analysis.

    PubMed

    Mies, C; Houldsworth, J; Chaganti, R S

    1991-02-01

    Tissues stored as paraffin blocks are a potential source of DNA for retrospective clinicogenetic analysis. To assess the feasibility of Southern blot analysis, DNA extracted from paraffin blocks was compared with DNA obtained from fresh-frozen controls of the same tissues. Sections 50-100 microns thick cut from paraffin blocks of 11 normal tissues, 18 lymphoid lesions, and 9 gastric carcinoma samples were deparaffinized and incubated at 45 degrees C for 48 to 72 h in a sodium dodecyl sulfate (SDS)/proteinase K solution. Following organic extraction, alcohol precipitation, restriction endonuclease digestion, and gel electrophoresis, DNA was transferred to nylon membranes. 32P-labelled DNA probes for the immunoglobulin heavy-chain locus and T-cell receptor beta-chain gene were hybridized to the normal tissue and lymphoid samples; the gastric cancers were probed for the HER-2/neu protooncogene. Intact DNA was obtained from the majority of formalin-fixed samples, yielding results qualitatively similar to those from fresh tissues. Degradation is the most significant problem in analyzing DNA extracted from paraffin blocks and compromises accurate quantitation. DNA analysis using paraffin-embedded tissue has potential clinical and research applications and may be a particularly useful way to study gene abnormalities in unusual tumors infrequently available as fresh specimens.

  18. Fire activity and severity in the western US vary along proxy gradients representing fuel amount and fuel moisture.

    PubMed

    Parks, Sean A; Parisien, Marc-André; Miller, Carol; Dobrowski, Solomon Z

    2014-01-01

    Numerous theoretical and empirical studies have shown that wildfire activity (e.g., area burned) at regional to global scales may be limited at the extremes of environmental gradients such as productivity or moisture. Fire activity, however, represents only one component of the fire regime, and no studies to date have characterized fire severity along such gradients. Given the importance of fire severity in dictating ecological response to fire, this is a considerable knowledge gap. For the western US, we quantify relationships between climate and the fire regime by empirically describing both fire activity and severity along two climatic water balance gradients, actual evapotranspiration (AET) and water deficit (WD), that can be considered proxies for fuel amount and fuel moisture, respectively. We also concurrently summarize fire activity and severity among ecoregions, providing an empirically based description of the geographic distribution of fire regimes. Our results show that fire activity in the western US increases with fuel amount (represented by AET) but has a unimodal (i.e., humped) relationship with fuel moisture (represented by WD); fire severity increases with fuel amount and fuel moisture. The explicit links between fire regime components and physical environmental gradients suggest that multivariable statistical models can be generated to produce an empirically based fire regime map for the western US. Such models will potentially enable researchers to anticipate climate-mediated changes in fire recurrence and its impacts based on gridded spatial data representing future climate scenarios. PMID:24941290

  19. Interdecadal modulation on the relationship between ENSO and typhoon activity during the late season in the western North Pacific

    NASA Astrophysics Data System (ADS)

    Zhao, Haikun; Wang, Chunzai

    2016-07-01

    The present study identifies an interdecadal modulation of the Pacific decadal oscillation (PDO) on the relationship between El Niño-Southern oscillation (ENSO) and typhoon activity during the late season (October-December) in the western North Pacific. The PDO is uncorrelated with ENSO during the warm phase of 1979-1997, while the PDO is positively correlated with ENSO during the cold phase of 1998-2012. Further analyses show that the warm phase is associated with the reduced ENSO-typhoon activity relationship and more typhoons, whereas the cold phase is corresponded to the enhanced ENSO-typhoon activity relationship and fewer typhoons. These variations are mainly manifested by a significant difference of typhoon activity in the southeastern part of the western North Pacific. Moreover, the change of ENSO-typhoon relationship is largely due to changes in large-scale environmental conditions especially from low-level vorticity and vertical wind shear between the two phases, which are related to the changes in tropical Indo-Pacific sea surface temperature. The study implies that the phase of the PDO should be taken into account when ENSO is used as a predictor for predicting typhoon activity in the western North Pacific.

  20. Fire Activity and Severity in the Western US Vary along Proxy Gradients Representing Fuel Amount and Fuel Moisture

    PubMed Central

    Parks, Sean A.; Parisien, Marc-André; Miller, Carol; Dobrowski, Solomon Z.

    2014-01-01

    Numerous theoretical and empirical studies have shown that wildfire activity (e.g., area burned) at regional to global scales may be limited at the extremes of environmental gradients such as productivity or moisture. Fire activity, however, represents only one component of the fire regime, and no studies to date have characterized fire severity along such gradients. Given the importance of fire severity in dictating ecological response to fire, this is a considerable knowledge gap. For the western US, we quantify relationships between climate and the fire regime by empirically describing both fire activity and severity along two climatic water balance gradients, actual evapotranspiration (AET) and water deficit (WD), that can be considered proxies for fuel amount and fuel moisture, respectively. We also concurrently summarize fire activity and severity among ecoregions, providing an empirically based description of the geographic distribution of fire regimes. Our results show that fire activity in the western US increases with fuel amount (represented by AET) but has a unimodal (i.e., humped) relationship with fuel moisture (represented by WD); fire severity increases with fuel amount and fuel moisture. The explicit links between fire regime components and physical environmental gradients suggest that multivariable statistical models can be generated to produce an empirically based fire regime map for the western US. Such models will potentially enable researchers to anticipate climate-mediated changes in fire recurrence and its impacts based on gridded spatial data representing future climate scenarios. PMID:24941290

  1. Attribution of extreme events in the western US to human activities

    NASA Astrophysics Data System (ADS)

    Mera, R. J.

    2015-12-01

    A project to investigate the role of human activities on the changing nature of extreme events in the western US began as part of a CLIVAR-sponsored Postdocs Applying Climate Expertise (PACE) project. The climate institution was the Oregon State University and the application partner was the Oregon Department of Land Conservation and Development (DLCD). DLCD was interested in the changes in weather extremes in the Pacific Northwest, specifically extreme rainfall, flooding, and droughts. The project employs very large ensembles of regional model simulations through volunteer computing resources and allows for probabilistic event attribution (PEA), an important climate research technique. The model was found to have good representation of atmospheric rivers, a major source of extreme precipitation in the Pacific Northwest. The model domain also encompasses California and Nevada. One of the studies focused on attribution of extreme heat in relation to vulnerable populations in California's Central Valley, where heat waves have become progressively more severe due to increasing nighttime temperatures. Specifically, we found that that (1) simulations of the hottest summer days during the 2000s were twice as likely to occur using observed levels of greenhouse gases than in a counterfactual world without major human activities, (2) detrimental impacts of heat on public health-relevant variables, such as the number of days above 40°C, can be quantified and attributed to human activities using PEA, and (3) PEA can serve as a tool for addressing climate justice concerns of populations within developed nations. The research conducted through the PACE program has also provided a framework for a pioneering climate attribution study at the Union of Concerned Scientists (UCS). The UCS project takes advantage of new research that shows that nearly two-thirds of carbon pollution released into the atmosphere, reported as carbon dioxide equivalent with hundred-year global warming

  2. Active tectonics and Quaternary landscape evolution across the western Panama block, Costa Rica, Central America

    NASA Astrophysics Data System (ADS)

    Marshall, Jeffrey Scott

    Three aspects of active tectonism are examined across central Costa Rica: (1) fault kinematics; (2) volcanic arc retreat; and (3) spatially variable coastal uplift. Diffuse faulting along the Central Costa Rica Deformed Belt (CCRDB) defines the western margin of the Panama block and aligns with the rough-smooth boundary (RSB) on the subducting Cocos plate. Sub-horizontal subduction of rough, hotspot thickened crust (Cocos Ridge and seamounts) shifts active shortening into the volcanic arc along the CCRDB. Mesoscale faults express variable kinematics across three domains: transtension in the forearc, transcurrent motion across the volcanic arc, and transpression in the back arc. Fault kinematics agree with seismicity and GPS data, and isotopic ages confirm that faulting postdates the late Neogene onset of shallow subduction. Stratigraphic correlation augmented by 40Ar/39Ar dating constrain the timing of Quaternary arc migration from the Neogene Aguacate range to the modern Cordillera Central. The Valle Central basin, between the cordilleras, filled with thick sequences of lavas, pyroclastic flows, and lahars. Middle Pleistocene drainage capture across the Aguacate arc linked the Valle Central with the Pacific slope and ash flows descended onto the coastal Orotina debris fan. Arc retreat reflects slab shallowing and enhanced tectonic erosion as rough crust entered the subduction zone. Differing subduction parameters across the RSB (crustal age, slab dip, roughness) produce marked contrasts in coastal tectonism. Varying uplift rates across coastal faults reflect sub-horizontal subduction of seamount roughness. Three groups (I--III) of fluvial terraces are correlated along the coast by isotopic ages and geomorphic characteristics. Base level fluctuations and terrace genesis reflect interaction between eustatic sea level and spatially variable rock uplift. Low uplift rates (north of RSB), yield one surface per terrace group, whereas moderate rates (south of RSB

  3. Seismicity on the western Greenland Ice Sheet: Surface fracture in the vicinity of active moulins

    DOE PAGES

    Carmichael, Joshua D.; Joughin, Ian; Behn, Mark D.; Das, Sarah; King, Matt A.; Stevens, Laura; Lizarralde, Dan

    2015-06-25

    We analyzed geophone and GPS measurements collected within the ablation zone of the western Greenland Ice Sheet during a ~35 day period of the 2011 melt season to study changes in ice deformation before, during, and after a supraglacial lake drainage event. During rapid lake drainage, ice flow speeds increased to ~400% of winter values, and icequake activity peaked. At times >7 days after drainage, this seismicity developed variability over both diurnal and longer periods (~10 days), while coincident ice speeds fell to ~150% of winter values and showed nightly peaks in spatial variability. Approximately 95% of all detected seismicitymore » in the lake basin and its immediate vicinity was triggered by fracture propagation within near-surface ice (<330 m deep) that generated Rayleigh waves. Icequakes occurring before and during drainage frequently were collocated with the down flow (west) end of the primary hydrofracture through which the lake drained but shifted farther west and outside the lake basin after the drainage. We interpret these results to reveal vertical hydrofracture opening and local uplift during the drainage, followed by enhanced seismicity and ice flow on the downstream side of the lake basin. This region collocates with interferometric synthetic aperture radar-measured speedup in previous years and could reflect the migration path of the meltwater supplied to the bed by the lake. The diurnal seismic signal can be associated with nightly reductions in surface melt input that increase effective basal pressure and traction, thereby promoting elevated strain in the surficial ice.« less

  4. K-T magmatism of western Rajasthan, India: Manifestation of Reunion plume activity or extensional lithospheric tectonics?

    NASA Astrophysics Data System (ADS)

    Sharma, K.

    2004-12-01

    Seychelles microcontinent from India, sedimentary basin development in western Rajasthan and the alkaline magmatism of Mundwara, Sarnu-Dandali and elsewhere are considered to be the products of Reunion plume activity in western India. However, basin development began in western Rajasthan in the Jurassic period and no plume has been suggested for this. The continual extensional tectonic regime caused deep fractures in the continental and oceanic lithosphere. The Cambay-Sanchor-Barmer rift developed in continental lithosphere. The Mundwara, Sarnu-Dandali and Barmer magmatism with nephelinite-carbonatite affinity at the basin margin represents a typical rift-tectonic setting. The tectonic setting and crustal development during the K-T period in western Rajasthan represents an extensional tectonic regime rather than the manifestation of Reunion plume activity.

  5. Seasonal Activity, Density, and Collection Efficiency of the Blacklegged Tick (Ixodes scapularis) (Acari: Ixodidae) in Mid-Western Pennsylvania.

    PubMed

    Simmons, T W; Shea, J; Myers-Claypole, M A; Kruise, R; Hutchinson, M L

    2015-11-01

    Although Pennsylvania has recently reported the greatest number of Lyme disease cases in the United States, with the largest increase for PA occurring in its western region, the population biology of the blacklegged tick (Ixodes scapularis Say) has not been adequately characterized in western PA. We studied the seasonal activity of host-seeking I. scapularis larvae, nymphs, and adults in mid-western PA over the course of a year, including a severe winter, and determined their absolute densities and collection efficiencies using replicated mark-release-recapture or removal methods. Our results are compared to those from similar studies conducted in the highly Lyme disease endemic Hudson Valley region of southeastern New York State. The seasonal activity of I. scapularis was intermediate between patterns observed in the coastal northeastern and upper Midwestern United States. Only one peak of larval activity was observed, which was later than the major peak in the Midwest, but earlier than in the northeast. Seasonal synchrony of larvae and nymphs was similar to the northeast, but the activity peaks were much closer together, although not completely overlapping as in the Midwest. Pre- and postwinter relative densities of questing adult I. scapularis were not significantly different from one another. The absolute densities and collection efficiencies of larvae, nymphs, and adults were comparable to results from classic research conducted at the Louis Calder Center in Westchester County, NY. We conclude that the population biology of I. scapularis in mid-western PA is similar to southeastern NYS contributing to a high acarological Lyme disease risk. PMID:26336271

  6. Western Samoa.

    PubMed

    1985-12-01

    This discussion of Western Samoa, which lies 2575 km northeast of Auckland, New Zealand, focuses on the following: geography; the people; history; government; political conditions; the economy; foreign relations; and relations the US. The population of Western Samoa, as of 1985, totals 163,000 with an annual growth rate of 0.9%. The infant mortality rate is 13/1000; life expectancy is 65 years. The main islands are formed ranges of extinct volcanoes. Volcanic activity last occurred in 1911. More than 2000 years age, waves of Polynesians migrated from Southeast Asia to the Samoan Islands. Samoans are the 2nd largest Polynesian group, after the Maoris of New Zealand, and speak a Polynesian dialect. Samoans have tended to retain their traditional ways despite exposure to European influence for more than 150 years. Most Samoans live within the traditional social system based on an extended family group, headed by a chief. Western Samoans are Christian. Education is free but not compulsory. In 1967, 95% of the children of primary school age attended school. From 1947 to 1961, a series of constitutional advances, assisted by visits from UN missions, brought Western Samoa from dependent status to self-government and finally to independence. The 1960 constitution is based on the British pattern of parliamentary democracy, modified to take Samoan customs into account. The present head of state holds his position for life. Future heads of state will be elected by the Legislative Assembly for 5-year terms. The Parliament consists of the Legislative Assembly and the head of state. The Supreme Court is the superior court of record and has full jurisdiction in civil, criminal, and constitutional matters. The "matai" of chief system still dominates the politics of Western Samoa, although several political parties have been formed and seem to be taking root. The "matai" system is a predominantly conservative force but does provide for change. Western Samoa is predominantly

  7. Igneous activity and related ore deposits in the western and southern Tushar Mountains, Marysvale volcanic field, west-central Utah

    USGS Publications Warehouse

    Steven, Thomas A.

    1984-01-01

    PART A: Igneous activity in the Marysvale volcanic field of western Utah can be separated into many episodes of extrusion, intrusion, and hydrothermal activity. The rocks of the western Tushar Mountains, near the western part of the volcanic field, include intermediate-composition, calc-alkalic volcanic rocks erupted from scattered volcanoes in Oligocene through earliest Miocene time and related monzonitic intrusions emplaced 24-23 m.y. ago. Beginning 22-21 m.y. ago and extending through much of the later Cenozoic, a bimodal basalt-rhyolite assemblage was erupted widely throughout the volcanic field. Only volcanic and intrusive rocks belonging to the rhyolitic end member of this bimodal assemblage are present in the western Tushar Mountains; most of these rocks either fill the Mount Belknap caldera (19 m.y. old) or are part of the rhyolite of Gillies Hill (9---8 m.y. old). Episodic hydrothermal activity altered and mineralized rocks at many places in the western Tushar Mountains during Miocene time. The earliest activity took place in and adjacent to monzonitic calcalkalic intrusions emplaced in the vicinity of Indian Creek and Cork Ridge. These rocks were widely propylitized, and gold-bearing quartz-pyrite-carbonate veins formed in local fractures. Hydrothermal activity associated with the Mount Belknap caldera mobilized and redeposited uranium contained in the caldera-fill rocks and formed primary concentrations of lithophile elements (including molybdenum and uranium) in the vicinity of intrusive bodies. Hydrothermal activity associated with the rhyolite of Gillies Hill altered and mineralized rocks at several places along the fault zone that marks the western margin of the Tushar Mountains; the zoned alunite and gold deposits at Sheep Rock, the gold deposit at the Sunday Mine, and an alunite deposit near Indian Creek were thus produced. Resetting of isotopic ages suggests that another center of hydrothermally altered rocks associated with a buried pluton about

  8. Hydrologic Vulnerability of Western US Rangelands in the Wake of Woodland Encroachment and Increasing Wildfire Activity

    NASA Astrophysics Data System (ADS)

    Williams, C. J.; Pierson, F. B.; Al-Hamdan, O. Z.; Kormos, P. R.

    2013-12-01

    Pinyon and juniper woodlands have dramatically increased their range in the past 150 years and currently occupy more than 30 million ha of the western US. Range expansion has primarily occurred through encroachment into sagebrush rangelands. Woodland expansion and infill on western rangelands have altered the ecological structure and function of these ecosystems and have made much of the western US prone to large severe wildfires. Early-succession woodlands are now burning in large, high-severity wildfires due to heavy woody-fuel loading and extensive horizontal-to-vertical fuel connectivity. Tree infill on late-succession woodlands coupled with extreme fire weather has increased the occurrence of large, high-severity woodland fires in recent decades. We investigated the effects of woodland encroachment and burning on hydrologic vulnerability at multiple woodlands and at a sagebrush rangeland in the early stages of woodland encroachment. Artificial rainfall and overland flow simulations were paired with vegetation and soil measures to evaluate ecohydrologic ramifications of woodland encroachment and burning at multiple spatial scales and over time. Our results provide insight into the ecohydrologic consequences of landscape-scale conversion of sagebrush rangelands to woodlands and the effects of increasing wildfire across this domain in the western US.

  9. Synthetic feeding stimulants enhance insecticide activity against western corn rootworm larvae, Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae)

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In behavioral bioassays, the addition of a synthetic feeding stimulant blend improved the efficacy of the insecticide thiamethoxam against neonate western corn rootworm, Diabrotica virgifera virgifera LeConte, larvae. In 4-h bioassays, the concentration of thiamethoxam required for 50% mortality (LC...

  10. Reappraisal of the 1887 Ligurian earthquake (western Mediterranean) from macroseismicity, active tectonics and tsunami modelling

    NASA Astrophysics Data System (ADS)

    Larroque, C.; Scotti, O.; Ioualalen, M.; Hassoun, V.; Migeon, S.

    2012-04-01

    Early in the morning, of February 23, 1887 a major damaging earthquake hit the towns along the Italian and French Riviera. The earthquake was followed by a tsunami wave with a maximum runup of 2 m near Imperia. At least 600 hundred people died, mainly due to building collapses. The "Ligurian earthquake" occurred at the junction between the Southern French-Italian Alps and the Ligurian Basin in the western Mediterranean. For such historical event, the epicentre and the equivalent magnitude are difficult to characterize with a high degree of precision, and the tectonic fault responsible for the earthquake is still debated today. The recent MALISAR marine geophysical survey allowed identifying a set of N60-70°E recent scarps at the foot of the northern Ligurian margin, revealing a large system of active faults. The scarps correspond to cumulative reverse faulting, with a minor strike-slip component, consistent with the present-day kinematics of earthquakes. Since we have also identified submarine failures in the time-range of the Ligurian earthquake we addressed the question of the submarine slide-induced tsunami. Nevertheless, the maximum volume involved by these submarine slides was in the range of 0.005 km3. Such a volume appears too small to trigger a tsunami with the observed extent and run-up characteristics. Therefore, we propose that the rupture of fault segments belonging to the 80 km-long northern Ligurian Faults system is the source of the 1887 Ligurian earthquake. We investigate the macroseismic data from the historical databases SISFRANCE-08 and DBMI-04 using several models of intensity attenuation with distance and focal depth. Modelling results are consistent with the location offshore, indicating an epicentre around 43.70°-43.78°N and 7.81°-8.07°E with a magnitude Mw in the range of 6.3-7.5. A refinement of this range of magnitude is discussed in the light of the tsunami modelling. Numerous earthquake sources scenarios have been tested with

  11. Protein blot assays specific for the discrimination of the centromere autoantigen, CENP-A, from human cells.

    PubMed

    Billings, P B; Martinez, A; Haselby, J A; Hoch, S O

    1993-09-01

    The Western or protein blot has proven to be a valuable resource in detecting discrete, immunoreactive antigen targets associated with a variety of autoimmune diseases. As the roster of autoantigens has expanded, it has become increasingly common to tailor specific gel or blot conditions to a particular polypeptide antigen. Two such assays are reported here as applied to the fractionation and visualization of human centromere protein (CENP-A), a centromere autoantigen associated with the rheumatic disease, systemic sclerosis. The centromere antigens are effectively solubilized in the presence of 1 M MgCl2 to allow for further purification. CENP-A copurifies with the histone proteins, primarily H3 and H4. The two CENP-A-specific protein blot assays separate CENP-A from the histone proteins and enhance CENP-A immunoreactivity. The first assay is based on the use of acid-urea gels with a Triton X-100 concentration chosen to maximize separation of CENP-A from all the histones. The second assay is based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis to differentiate two very basic proteins of similar molecular weight, namely CENP-A and histone H3. For each gel system, a selective choice of associated immunoblot parameters allows for the reproducible discrimination of the CENP-A antigen. PMID:8223400

  12. Lectin blot studies on proteins of Myxobolus cerebralis, the causative agent of whirling disease.

    PubMed

    Knaus, Martin; El-Matbouli, Mansour

    2005-07-18

    It is known that Myxobolus cerebralis antigens, both surficial and secreted, are key modulators for, or targets of, host immune system compounds. We undertook SDS-PAGE glycoprotein characterisation of M. cerebralis developmental stages isolated from infected rainbow trout and Western blot analyses using selected biotin-labelled plant lectins (GSA-I, PHA-E, SJA, GSA-II) and anti-triactinomyxon polyclonal antibodies. Glycoproteins were isolated with lectin-affinity chromatography, and prominent bands were characterised by matrix-assisted laser desorption/ionisation-mass spectrometry (MALDI/MS). We identified glycoproteins of M. cerebralis myxospores that contained carbohydrate motifs reactive with Phaseolus vulgaris erythroagglutinin (proteins 20 to 209 kDa, PHA-E), Sophora japonica agglutinin (proteins 7 to 70 kDa, SJA), Griffonia simplicifolia Agglutinin I (proteins 10 to 209 kDa, GSA-I) and G. simplicifolia Agglutinin II (proteins 5 to 40 kDa, GSA-II). Mcgp33, a glycoprotein isolated by lectin-affinity chromatography, was reactive with SJA (about 33 kDa). Antiserum produced against M. cerebralis triactinomyxons was found to have differences in the antigenicity of isolated glycoproteins from both M. cerebralis myxospores and actinospores. We also demonstrated modified antigen expression, especially involving the glycoprotein Mcgp33, in different developmental stages of M. cerebralis. PMID:16119891

  13. A sensitive non-radioactive northern blot method to detect small RNAs.

    PubMed

    Kim, Sang Woo; Li, Zhihua; Moore, Patrick S; Monaghan, A Paula; Chang, Yuan; Nichols, Mark; John, Bino

    2010-04-01

    The continuing discoveries of potentially active small RNAs at an unprecedented rate using high-throughput sequencing have raised the need for methods that can reliably detect and quantitate the expression levels of small RNAs. Currently, northern blot is the most widely used method for validating small RNAs that are identified by methods such as high-throughput sequencing. We describe a new northern blot-based protocol (LED) for small RNA (approximately 15-40 bases) detection using digoxigenin (DIG)-labeled oligonucleotide probes containing locked nucleic acids (LNA) and 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide for cross-linking the RNA to the membrane. LED generates clearly visible signals for RNA amounts as low as 0.05 fmol. This method requires as little as a few seconds of membrane exposure to outperform the signal intensity using overnight exposure of isotope-based methods, corresponding to approximately 1000-fold improvement in exposure-time. In contrast to commonly used radioisotope-based methods, which require freshly prepared and hazardous probes, LED probes can be stored for at least 6 months, facilitate faster and more cost-effective experiments, and are more environmentally friendly. A detailed protocol of LED is provided in the Supplementary Data. PMID:20081203

  14. Heterotrophic bacterial production and extracellular enzymatic activity in sinking particulate matter in the western North Pacific Ocean.

    PubMed

    Yamada, Namiha; Fukuda, Hideki; Ogawa, Hiroshi; Saito, Hiroaki; Suzumura, Masahiro

    2012-01-01

    Heterotrophic activities on sinking particulate matter (SPM) play an important role in SPM fluxes in the ocean. To demonstrate regional differences in heterotrophic activities on SPM, we measured heterotrophic bacterial production (HBP) in seawater (HBP(SW)) and SPM (HBP(SPM)) as well as potential extracellular enzyme activity (EEA) in SPM on a transect along 155°E in the western North Pacific Ocean in the subarctic (44°N), the Kuroshio Extension area (35°N), and the subtropical gyre (20°N). Depth-integrated HBP(SW) from the surface to 500 m was comparable between the locations, whereas HBP(SPM) at 44°N was substantially lower than at the other sites. We found the highest particulate organic carbon (POC) export flux and export efficiency to bathypelagic depths, and the lowest water temperatures, at 44°N. We found significant correlations between leucine aminopeptidase (LAPase) activity, β-glucosidase (BGase) activity, POC flux and particulate organic nitrogen flux. LAPase activity was two orders of magnitude higher than BGase activity, with a BGase:LAPase activity ratio of 0.027. There were no significant correlations between HBP and EEA in SPM except for lipase, and lipase activity was significantly correlated with temperature. We propose that hydrographic conditions are an important factor controlling heterotrophic bacterial activity and export efficiency of organic carbon to the deep ocean, as are the sources and abundance of SPM produced in the euphotic zone via primary production. PMID:23109933

  15. New insights into the active deformation of accretionary prisms: examples from the Western Makran, Iran

    NASA Astrophysics Data System (ADS)

    Penney, Camilla; Copley, Alex; Oveisi, Benham

    2016-04-01

    The Makran subduction zone, along the southern coasts of Iran and Pakistan, hosts one of the largest exposed accretionary wedges in the world. The western Makran has been characterised by a lack of shallow and thrust seismicity in both the instrumental and historical periods. The Mw 6.1 2013 Minab earthquake thus provides a rare opportunity to study the deformation of the accretionary wedge in the transition region between continent-continent collision, in the Zagros, and oceanic subduction, in the Makran. We study the source parameters and slip distribution of this earthquake using seismology, geodesy and field observations. We observe left-lateral strike-slip motion on a fault striking ENE-WSW; approximately perpendicular to the faults of the Minab-Zendan-Palami fault zone, the main structure previously thought to accommodate the right-lateral shear between the Zagros and the Makran. The fault that ruptured in 2013 is one of a series of approximately E-W striking left-lateral faults visible in the geology and geomorphology. These accommodate a velocity field equivalent to right-lateral shear on N-S striking planes by clockwise rotations about vertical axes. The longitudinal range of shear in the western Makran is likely to be controlled by the distance over which the underthrusting Arabian lithosphere deepens in the transition from continent-continent collision to oceanic subduction. The lack of observed megathrust seismicity in the western Makran has led to assertions that the convergence in this region may be aseismic, in contrast to the eastern Makran, which experienced an Mw8.1 earthquake in 1945. The right-lateral Sistan Suture Zone, which runs ~N-S along the Iran-Afghanistan border to the north of the Makran, appears to separate these regimes. However, right-lateral faulting is not observed south of ~27°N, within the wedge. The Minab earthquake and the 2013 Balochistan earthquake show that the Makran accretionary wedge is dominated by strike-slip faulting

  16. Recent Extreme Forest Fire Activity in Western Russia: Fire Danger Conditions, Fire Behavior and Smoke Transport

    NASA Astrophysics Data System (ADS)

    Stocks, B. J.; Fromm, M.; Goldammer, J.; Carr, R.; Sukhinin, A. I.

    2010-12-01

    During the summer of 2010, widespread forest and peatland fires in western Russia burned over hundreds of thousands of hectares, burning over croplands, destroying hundreds of homes, and directly causing the death of more than 50 people. Unprecedented drought conditions, combined with an extended heat wave, resulted in extreme fire danger conditions and explosive fire behavior in a region of Russia not noted for large fires. Several fires exhibited pyroconvection, injecting smoke directly into the upper troposphere and lower stratosphere, while deep-burning fires created major regional smoke problems. This smoke persisted in the heavily-populated areas around Moscow, exposing millions to high levels of ozone and particulate matter, and creating both immediate and longer-term health risks. This presentation will explore the drought conditions leading to the catastrophic fire behavior experienced in western Russia, and analyze fire behavior in terms of fuel consumption, smoke production, fire intensity levels, and pyroconvection. Impacts of regional and long-range smoke transport will also be discussed.

  17. Restoration of antibody binding to blotted meningococcal outer membrane proteins using various detergents.

    PubMed

    Wedege, E; Bryn, K; Frøholm, L O

    1988-10-01

    Restoration of IgG antibody binding to heat-denatured meningococcal outer membrane proteins has been studied on immunoblots with a series of 14 detergents. Nitrocellulose strips with the blotted proteins were incubated with the detergents and sera from human volunteers vaccinated with meningococcal membrane proteins. Zwitterionic and ionic detergents, containing substituted quarternary ammonium or amino groups with a minimum of 10 C atoms in the alkyl chain, restored the antigenicity of the serotype-specific class 2 porin protein. The concentrations of the Zwittergent detergents necessary for activation decreased with increasing alkyl chain length of the homologues. Only zwitterionic detergents renatured the class 1 protein. Both proteins were weakly antigenic in the presence of the nonionic detergents Triton X-100 and Tween 20. Meningococcal lipopolysaccharide restored antibody binding to the porin, but not to the class 1 protein. Similar concentrations of lipopolysaccharides from two other gram-negative bacteria had no effect.

  18. 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.

  19. Stratigraphic development and hydrothermal activity in the central western Cascade Range, Oregon

    SciTech Connect

    Cummings, M.L.; Bull, M.K. ); Pollock, J.M. ); Thompson, G.D. )

    1990-11-10

    Two volcanic sequences bounded by erosional unconformities compose the stratigraphy of the North Santiam mining district, Western Cascade Range, Oregon. Diorite, grandodiorite, and leucocratic quartz porphyry dikes, stocks, and sills intrude the breccias, flows, and tuffs of a volcanic center in the older Sardine Formation. Tourmaline-bearing breccia pipes are associated with the porphyritic granodiorite intrusions. An erosional unconformity separates the Sardine Formation from the overlying Elk Lake formation. The alteration patterns in the two formations are consistent with the development of hydrothermal systems during the eruption of each formation. However, the development of the two hydrothermal systems is separated by a period of erosion of the older volcanic pile. Early formation of mineralization that resembles porphyry copper deposits occurred within the Sardine Formation, and later, after eruption of the Elk Lake formation, epithermal veins and alteration developed along faults, fractures, and the margins of dikes in the Sardine Formation.

  20. RNase protection assays and RNA gel blots: a direct comparison of sensitivity.

    PubMed

    Higgs, D C; Colbert, J T

    1992-01-01

    RNase protection assays are commonly thought to be a more sensitive means of detecting and quantitating specific mRNAs than are RNA gel blots (Northern blots). We have directly compared the sensitivity of these two approaches by assaying for known amounts of in vitro synthesized beta-glucuronidase mRNA. With the probes and protocols employed here, the ability to detect a specific mRNA was similar whether RNase protection or RNA gel blot analyses were performed.

  1. Provenance and sediment-dispersal system in tectonically active rapidly evolving foreland basin, Western Interior

    SciTech Connect

    Khandaker, N.I.; Vondra, C.F.

    1989-03-01

    The Upper Cretaceous Frontier Formation, along the mobile edge of the Western Interior foreland basin, is composed mainly of clastic sediments and was deposited during the initial Late Cretaceous transgressive-regressive phases of the Western Interior seaway across Wyoming. The formation contains many persistent bentonite beds and several sandstone packages in its lower part and a thin, lenticular lithic wacke-polymictic conglomerate association at its upper contact (Torchlight Sandstone Member). Abundant granule to cobble-sized clasts of andesite, granite, chert, and quartzite are set in a poorly sorted sand-to-granule grade volcaniclastic matrix. There is a lithologic continuity of this volcaniclastic unit across the Bighorn Mountains into the Powder River basin. A high-energy distributary complex of sizable areal extent is invoked for the deposition of this linear conglomerate facies. Geochemical investigations of the whole-rock andesite clasts and bentonite allowed more precise definition of character, tectonic setting, and evolutionary stages of sedimentary distributive provinces. Bentonites and andesites are strongly enriched in strontium and barium, but only mildly enriched in heavy rare earth elements and high field-strength elements. These analyzed rocks have trace element characteristics similar in a general way to those of typical orogenic volcanics; they show some significant differences in detail. Composition of volcaniclasts and paleocurrent data indicate a proximal sediment source for the extrabasinal detritus within the Frontier Formation. The possibility of a contribution from a Mesozoic volcanic center in the neighborhood of southwestern Montana is strongly favored. The products of this volcanism constitute an assemblage of deep crustal to mantle( ) derived rocks, and their composition record time-integrated enrichment in light over heavy rare earth elements.

  2. Transition from collision to subduction in Western Greece: the Katouna-Stamna active fault system and regional kinematics

    NASA Astrophysics Data System (ADS)

    Pérouse, E.; Sébrier, M.; Braucher, R.; Chamot-Rooke, N.; Bourlès, D.; Briole, P.; Sorel, D.; Dimitrov, D.; Arsenikos, S.

    2016-06-01

    Transition from subduction to collision occurs in Western Greece and is accommodated along the downgoing plate by the Kefalonia right-lateral fault that transfers the Hellenic subduction front to the Apulian collision front. Here we present an active tectonic study of Aitolo-Akarnania (Western Greece) that highlights how such a transition is accommodated in the overriding plate. Based on new multi-scale geomorphic and tectonic observations, we performed an accurate active fault trace mapping in the region, and provide evidence for active normal and left-lateral faulting along the Katouna-Stamna Fault (KSF), a 65-km-long NNW-striking fault system connecting the Amvrakikos Gulf to the Patras Gulf. We further show that the Cenozoic Hellenide thrusts located west of the KSF are no longer active, either in field observation or in GPS data, leading us to propose that the KSF forms the northeastern boundary of a rigid Ionian Islands-Akarnania Block (IAB). Cosmic ray exposure measurements of 10Be and 36Cl were performed on a Quaternary alluvial fan offset along the KSF (~50 m left-lateral offset). A maximum abandonment age of ~12-14 ka for the alluvial fan surface can be determined, giving an estimated KSF minimum geological left-lateral slip rate of ~4 mm year-1, in agreement with high GPS slip rates (~10 mm year-1). Despite this high slip rate, the KSF is characterized by subdued morphological evidence of tectonic activity, a gypsum-breccia bedrock and a low level of seismicity, suggesting a dominantly creeping behavior for this fault. Finally, we discuss how the IAB appears to have been progressively individualized during the Pleistocene (younger than ~1.5 Ma).

  3. Chronic consumption of a western diet induces robust glial activation in aging mice and in a mouse model of Alzheimer's disease.

    PubMed

    Graham, Leah C; Harder, Jeffrey M; Soto, Ileana; de Vries, Wilhelmine N; John, Simon W M; Howell, Gareth R

    2016-01-01

    Studies have assessed individual components of a western diet, but no study has assessed the long-term, cumulative effects of a western diet on aging and Alzheimer's disease (AD). Therefore, we have formulated the first western-style diet that mimics the fat, carbohydrate, protein, vitamin and mineral levels of western diets. This diet was fed to aging C57BL/6J (B6) mice to identify phenotypes that may increase susceptibility to AD, and to APP/PS1 mice, a mouse model of AD, to determine the effects of the diet in AD. Astrocytosis and microglia/monocyte activation were dramatically increased in response to diet and was further increased in APP/PS1 mice fed the western diet. This increase in glial responses was associated with increased plaque burden in the hippocampus. Interestingly, given recent studies highlighting the importance of TREM2 in microglia/monocytes in AD susceptibility and progression, B6 and APP/PS1 mice fed the western diet showed significant increases TREM2+ microglia/monocytes. Therefore, an increase in TREM2+ microglia/monocytes may underlie the increased risk from a western diet to age-related neurodegenerative diseases such as Alzheimer's disease. This study lays the foundation to fully investigate the impact of a western diet on glial responses in aging and Alzheimer's disease. PMID:26888450

  4. Chronic consumption of a western diet induces robust glial activation in aging mice and in a mouse model of Alzheimer’s disease

    PubMed Central

    Graham, Leah C.; Harder, Jeffrey M.; Soto, Ileana; de Vries, Wilhelmine N.; John, Simon W. M.; Howell, Gareth R.

    2016-01-01

    Studies have assessed individual components of a western diet, but no study has assessed the long-term, cumulative effects of a western diet on aging and Alzheimer’s disease (AD). Therefore, we have formulated the first western-style diet that mimics the fat, carbohydrate, protein, vitamin and mineral levels of western diets. This diet was fed to aging C57BL/6J (B6) mice to identify phenotypes that may increase susceptibility to AD, and to APP/PS1 mice, a mouse model of AD, to determine the effects of the diet in AD. Astrocytosis and microglia/monocyte activation were dramatically increased in response to diet and was further increased in APP/PS1 mice fed the western diet. This increase in glial responses was associated with increased plaque burden in the hippocampus. Interestingly, given recent studies highlighting the importance of TREM2 in microglia/monocytes in AD susceptibility and progression, B6 and APP/PS1 mice fed the western diet showed significant increases TREM2+ microglia/monocytes. Therefore, an increase in TREM2+ microglia/monocytes may underlie the increased risk from a western diet to age-related neurodegenerative diseases such as Alzheimer’s disease. This study lays the foundation to fully investigate the impact of a western diet on glial responses in aging and Alzheimer’s disease. PMID:26888450

  5. Chronic consumption of a western diet induces robust glial activation in aging mice and in a mouse model of Alzheimer's disease.

    PubMed

    Graham, Leah C; Harder, Jeffrey M; Soto, Ileana; de Vries, Wilhelmine N; John, Simon W M; Howell, Gareth R

    2016-02-18

    Studies have assessed individual components of a western diet, but no study has assessed the long-term, cumulative effects of a western diet on aging and Alzheimer's disease (AD). Therefore, we have formulated the first western-style diet that mimics the fat, carbohydrate, protein, vitamin and mineral levels of western diets. This diet was fed to aging C57BL/6J (B6) mice to identify phenotypes that may increase susceptibility to AD, and to APP/PS1 mice, a mouse model of AD, to determine the effects of the diet in AD. Astrocytosis and microglia/monocyte activation were dramatically increased in response to diet and was further increased in APP/PS1 mice fed the western diet. This increase in glial responses was associated with increased plaque burden in the hippocampus. Interestingly, given recent studies highlighting the importance of TREM2 in microglia/monocytes in AD susceptibility and progression, B6 and APP/PS1 mice fed the western diet showed significant increases TREM2+ microglia/monocytes. Therefore, an increase in TREM2+ microglia/monocytes may underlie the increased risk from a western diet to age-related neurodegenerative diseases such as Alzheimer's disease. This study lays the foundation to fully investigate the impact of a western diet on glial responses in aging and Alzheimer's disease.

  6. Hydrothermal fluids circulation and travertine deposition in an active tectonic setting: Insights from the Kamara geothermal area (western Anatolia, Turkey)

    NASA Astrophysics Data System (ADS)

    Brogi, Andrea; Alçiçek, M. Cihat; Yalçıner, Cahit Çağlar; Capezzuoli, Enrico; Liotta, Domenico; Meccheri, Marco; Rimondi, Valentina; Ruggieri, Giovanni; Gandin, Anna; Boschi, Chiara; Büyüksaraç, Aydin; Alçiçek, Hülya; Bülbül, Ali; Baykara, Mehmet Oruç; Shen, Chuan-Chou

    2016-06-01

    Coexistence of thermal springs, travertine deposits and tectonic activity is a recurring feature for most geothermal areas. Although such a certainty, their relationships are debated mainly addressing on the role of the tectonic activity in triggering and controlling fluids flow and travertine deposition. In this paper, we present the results of an integrated study carried out in a geothermal area located in western Anatolia (Turkey), nearby the well-known Pamukkale area (Denizli Basin). Our study focused on the relationships among hydrothermal fluids circulation, travertine deposition and tectonic activity, with particular emphasis on the role of faults in controlling fluids upwelling, thermal springs location and deposition of travertine masses. New field mapping and structural/kinematics analyses allowed us to recognize two main faults systems (NW- and NE-trending), framed in the Neogene-Quaternary extensional tectonic evolution of western Anatolia. A geo-radar (GPR) prospection was also provided in a key-area, permitting us to reconstruct a buried fault zone and its relationships with the development of a fissure-ridge travertine deposit (Kamara fissure-ridge). The integration among structural and geophysical studies, fluids inclusion, geochemical, isotopic data and 230 Th/238 U radiometric age determination on travertine deposits, depict the characteristics of the geothermal fluids and their pathway, up to the surface. Hydrological and seismological data have been also taken in account to investigate the relation between local seismicity and fluid upwelling. As a main conclusion we found strict relationships among tectonic activity, earthquakes occurrence, and variation of the physical/chemical features of the hydrothermal fluids, presently exploited at depth, or flowing out in thermal springs. In the same way, we underline the tectonic role in controlling the travertine deposition, making travertine (mainly banded travertine) a useful proxy to reconstruct the

  7. Imaging active faults in a region of distributed deformation from joint focal mechanism and hypocenter clustering: Application to western Iberia

    NASA Astrophysics Data System (ADS)

    Custodio, S.; Lima, V.; Vales, D.; Carrilho, F.; Cesca, S.

    2015-12-01

    Mainland Portugal, on the SW edge of the European continent, is located directly north of the boundary between the Eurasian and Nubian plates. It lies in a region of slow lithospheric deformation, which has generated some of the largest earthquakes in Europe, both intraplate (mainland) and interplate (offshore). The seismicity of mainland Portugal and its adjacent offshore has been repeatedly classified as diffuse. We analyse the instrumental earthquake catalog for western Iberia, enriched with data from recent dense broadband deployments. We show that although the plate boundary south of Portugal is diffuse, in that deformation is accommodated along several distributed faults rather than along one long linear plate boundary, the seismicity itself is not diffuse. Rather, when located using high quality data, earthquakes collapse into well-defined clusters and lineations. We then present a new joint focal mechanism and hypocenter cluster algorithm that is able to extract coherent information between hypocenter locations and focal mechanisms. We apply the method to the Azores-western Mediterranean region, with emphasis on western Iberia. In addition to identifying well-known seismo-tectonic features, the joint clustering algorithm identifies eight new clusters of earthquakes with a good match between the directions of epicentre lineations and focal mechanism fault planes. These clusters may signal single active faults or wider fault zones accommodating a consistent type of faulting. Mainland Portugal is dominated by strike-slip faulting, consistent with the NNE-SSW and WNW-ESE oriented lineations. The region offshore SW Iberia displays clusters that are either predominantly strike-slip or reverse, indicating slip partitioning. This work shows that the study of low-magnitude earthquakes using dense seismic deployments is a powerful tool to study lithospheric deformation in slowly deforming regions, where high-magnitude earthquakes occur with long recurrence intervals.

  8. Depopulation of rural landscapes exacerbates fire activity in the western Amazon.

    PubMed

    Uriarte, María; Pinedo-Vasquez, Miquel; DeFries, Ruth S; Fernandes, Katia; Gutierrez-Velez, Victor; Baethgen, Walter E; Padoch, Christine

    2012-12-26

    Destructive fires in Amazonia have occurred in the past decade, leading to forest degradation, carbon emissions, impaired air quality, and property damage. Here, we couple climate, geospatial, and province-level census data, with farmer surveys to examine the climatic, demographic, and land use factors associated with fire frequency in the Peruvian Amazon from 2000 to 2010. Although our results corroborate previous findings elsewhere that drought and proximity to roads increase fire frequency, the province-scale analysis further identifies decreases in rural populations as an additional factor. Farmer survey data suggest that increased burn scar frequency and size reflect increased flammability of emptying rural landscapes and reduced capacity to control fire. With rural populations projected to decline, more frequent drought, and expansion of road infrastructure, fire risk is likely to increase in western Amazonia. Damage from fire can be reduced through warning systems that target high-risk locations, coordinated fire fighting efforts, and initiatives that provide options for people to remain in rural landscapes. PMID:23236144

  9. Study of thunderstorm activity connection with weather system structures over the North-Western Pacific Ocean

    NASA Astrophysics Data System (ADS)

    Permyakov, M. S.; Cherneva, N. V.; Shevtsov, B. M.; Potalova, E. Yu.; Holzworth, Robert

    2014-11-01

    Applying the data of VLF direction finder receiving station of IKIR FEB RAS, included into the World Wide Lightning Location Network (WWLLN), the paper investigates the relations of field characteristics of recorded lightning discharges in the north-western part of the Pacific ocean with field characteristics of weather formation meteorological elements, evaluated according to the data of Earth remote sounding from satellites. On the example of separate tropical cyclones (TC) for 2012-2013, the relation of lightning discharge frequency and density with spatial distribution of driving wind whirl is shown. TC structure evolution is traced in cloudiness fields, driving wind whirl, and lightning discharge distribution. This publication is based on work supported by a grant from the U.S. Civilian Research and Development Foundation (RUG1-7084-PA-13) with funding from the United States Department of State. The opinions, findings and conclusions stated herein are those of the authors and do not necessarily reflect those of CRDF Global or the United States Department of State.

  10. Depopulation of rural landscapes exacerbates fire activity in the western Amazon.

    PubMed

    Uriarte, María; Pinedo-Vasquez, Miquel; DeFries, Ruth S; Fernandes, Katia; Gutierrez-Velez, Victor; Baethgen, Walter E; Padoch, Christine

    2012-12-26

    Destructive fires in Amazonia have occurred in the past decade, leading to forest degradation, carbon emissions, impaired air quality, and property damage. Here, we couple climate, geospatial, and province-level census data, with farmer surveys to examine the climatic, demographic, and land use factors associated with fire frequency in the Peruvian Amazon from 2000 to 2010. Although our results corroborate previous findings elsewhere that drought and proximity to roads increase fire frequency, the province-scale analysis further identifies decreases in rural populations as an additional factor. Farmer survey data suggest that increased burn scar frequency and size reflect increased flammability of emptying rural landscapes and reduced capacity to control fire. With rural populations projected to decline, more frequent drought, and expansion of road infrastructure, fire risk is likely to increase in western Amazonia. Damage from fire can be reduced through warning systems that target high-risk locations, coordinated fire fighting efforts, and initiatives that provide options for people to remain in rural landscapes.

  11. Antagonistic and plant growth activity of Trichoderma isolates of Western Himalayas.

    PubMed

    Joshi, B B; Bhatt, R P; Bahukhandi, D

    2010-11-01

    The genus Trichoderma is rapidly growing colonies bearing tufted or postulate, repeatedly branched conidiophores with lageniform phialides and hyaline or green conidia born in slimy heads. 62 isolates of Trichoderma species were isolated from different rhizospheric soil samples collected from different places located in Western Himalayas region. Out of these only two species were found i.e. Trichoderma hazianum and Trichoderma viride. Their efficacy against soil borne plant pathogens like Sclerotium rolfsii, Rhizoctonia solani and Sclerotinia sclerotiorum revealed that only three isolates amounting to 5% of the total collected isolates of this region were found highly antagonist. Among them 5% isolates were found against S. rolfsii, 13% isolates against R. solani, 10% against sclerotium caused above 80% inhibition of mycelial growth respectively. 6% isolates out of twenty seven utilized chitin by more than 80 and 16% isolates consumed cellulose by above 80% and therefore are producers of chitinase and cellulases. 58% isolates produced colonies having cottony texture and 41% produced dark green colonies. Pigmentation as observed from reverse side of the colony revealed that 70% of them did not produced pigment in the medium. Plant growth promotion measured as root and shoot lengths were significantly higher than in control. The maximum root length and shoot length were recorded when seeds were treated with isolates were recorded at Srinagar Garhwal was 4.70 and 4.75 cm out of all the isolates in which isolate recorded from Srinagar no 3 caused maximum percent seed germination which was significantly higher 79.49%.

  12. Depopulation of rural landscapes exacerbates fire activity in the western Amazon

    PubMed Central

    Uriarte, María; Pinedo-Vasquez, Miquel; DeFries, Ruth S.; Fernandes, Katia; Gutierrez-Velez, Victor; Baethgen, Walter E.; Padoch, Christine

    2012-01-01

    Destructive fires in Amazonia have occurred in the past decade, leading to forest degradation, carbon emissions, impaired air quality, and property damage. Here, we couple climate, geospatial, and province-level census data, with farmer surveys to examine the climatic, demographic, and land use factors associated with fire frequency in the Peruvian Amazon from 2000 to 2010. Although our results corroborate previous findings elsewhere that drought and proximity to roads increase fire frequency, the province-scale analysis further identifies decreases in rural populations as an additional factor. Farmer survey data suggest that increased burn scar frequency and size reflect increased flammability of emptying rural landscapes and reduced capacity to control fire. With rural populations projected to decline, more frequent drought, and expansion of road infrastructure, fire risk is likely to increase in western Amazonia. Damage from fire can be reduced through warning systems that target high-risk locations, coordinated fire fighting efforts, and initiatives that provide options for people to remain in rural landscapes. PMID:23236144

  13. Antagonistic and plant growth activity of Trichoderma isolates of Western Himalayas.

    PubMed

    Joshi, B B; Bhatt, R P; Bahukhandi, D

    2010-11-01

    The genus Trichoderma is rapidly growing colonies bearing tufted or postulate, repeatedly branched conidiophores with lageniform phialides and hyaline or green conidia born in slimy heads. 62 isolates of Trichoderma species were isolated from different rhizospheric soil samples collected from different places located in Western Himalayas region. Out of these only two species were found i.e. Trichoderma hazianum and Trichoderma viride. Their efficacy against soil borne plant pathogens like Sclerotium rolfsii, Rhizoctonia solani and Sclerotinia sclerotiorum revealed that only three isolates amounting to 5% of the total collected isolates of this region were found highly antagonist. Among them 5% isolates were found against S. rolfsii, 13% isolates against R. solani, 10% against sclerotium caused above 80% inhibition of mycelial growth respectively. 6% isolates out of twenty seven utilized chitin by more than 80 and 16% isolates consumed cellulose by above 80% and therefore are producers of chitinase and cellulases. 58% isolates produced colonies having cottony texture and 41% produced dark green colonies. Pigmentation as observed from reverse side of the colony revealed that 70% of them did not produced pigment in the medium. Plant growth promotion measured as root and shoot lengths were significantly higher than in control. The maximum root length and shoot length were recorded when seeds were treated with isolates were recorded at Srinagar Garhwal was 4.70 and 4.75 cm out of all the isolates in which isolate recorded from Srinagar no 3 caused maximum percent seed germination which was significantly higher 79.49%. PMID:21506476

  14. Geomorphic signatures of active tectonics in the Trans-Yamuna segment of the western Doon valley, northwest Himalaya, India

    NASA Astrophysics Data System (ADS)

    Philip, George; Sah, Madho P.

    Being involved in the late orogenic movements of the sub-Himalaya, the Doon valley and its Quaternary formations have received considerable attention from Earth scientists in the study of active tectonics and paleoseismic events. Study of aerial photographs and satellite data, and selected field checks not only confirmed neotectonic features already reported by various authors but also revealed the presence of more such features. In response to active tectonics, these features have affected very young terraces and Quaternary sediments in the Trans-Yamuna segment of the Doon valley in the western sub-Himalaya. In the present study, an attempt has been made to understand the neotectonic implications of these movements on landforms in and around Sataun-Sirmuri Tal. Ground evidence indicates that the area has experienced at least three major tectonic impulses since the generation of the Main Boundary Thrust. The major tectonic disturbances are most likely due to co-seismic activity along the ongoing Himalayan tectonic processes. In this paper, we discuss some of the strong geomorphic signatures, such as lineament and active fault traces, pressure ridges, sag ponds, alluvial fans, river terraces and finally landslides, which are indicative of active tectonics in this area. On the basis of the present-day geomorphic configuration of this sub-Himalayan basin, a possible evolutionary history is also presented.

  15. Active Deformation in the Greater Himalayan Zone in Western Nepal from Inversion of New (U-Th)/He Cooling Ages

    NASA Astrophysics Data System (ADS)

    Harvey, J. E.; Burbank, D.

    2015-12-01

    Much of the central Himalaya features an abrupt rise in mean elevation from ~1.5 km in the Lesser Himalaya to ~4-5 km Greater Himalaya and Tibetan Plateau. This physiographic transition is known as PT2, and is often interpreted as the surface expression of transport over a ramp in the Main Himalayan Thrust (MHT). In western Nepal, however, the same rise in elevation occurs over two distinct topographic steps (PT2-N and PT2-S). In previous work, Harvey et al. (2015) argue that this anomalous topography is the result of recent southward-migration of mid-crustal deformation along the MHT. Due to the seismogenic potential of the MHT it is important to constrain its geometry in the western Nepal seismic gap, which has not had a large earthquake in over 600 years. To test the above hypothesis, we perform [U-Th]/He dating on 39 apatite and 47 zircon samples collected along seven relief transects throughout western Nepal. We constrain exhumation histories by inverting these new cooling ages with the 3-D thermo-kinematic model Pecube. Five transects collected from the Greater Himalaya north of PT2-N are best fit by relatively rapid exhumation rates (~1-2 km/Myr) since ~4 Ma. The other two, collected from farther south near PT2-S, require rapid (~1-2 km/Myr) exhumation until around 8-11 Ma, followed by much slower (~0.1-0.2 km/Myr) exhumation until at least the late Pliocene. Assuming that exhumation rates reflect uplift rates, the rapid Plio-Pleistocene exhumation in the Greater Himalaya north of PT2-N suggest that this physiographic transition is similar to that at the foot of the Greater Himalaya in central Nepal. It follows that active deformation is occurring along a NW-trend as much as 100 km farther north than would be expected if simply projecting PT2 across western Nepal. This finding is consistent with transport over a more northerly MHT ramp or perhaps oblique slip along the recently identified, surface-breaking WNFZ. Although the geomorphology and microseismicity

  16. Antiviral activity and underlying molecular mechanisms of Matrine against porcine reproductive and respiratory syndrome virus in vitro.

    PubMed

    Sun, Na; Wang, Zhi-Wei; Wu, Cai-Hong; Li, E; He, Jun-Ping; Wang, Shao-Yu; Hu, Yuan-Liang; Lei, Hai-Min; Li, Hong-Quan

    2014-04-01

    Porcine reproductive and respiratory syndrome (PRRS), caused by porcine reproductive and respiratory syndrome virus (PRRSV), is an acute infectious disease. The prevalence of PRRS has made swine industry suffered huge financial losses. Matrine, a natural compound, has been demonstrated to possess anti-PRRSV activity in Marc-145 cells. However, the underlying molecular mechanisms were still unknown. The main objective of our study was to discuss the effect of Matrine on PRRSV N protein expression and PRRSV induced apoptosis. Indirect immunofluorescence assay (IFA) and Western blot were used to assess the effect of Matrine on N protein expression. Apoptosis was analyzed by fluorescence staining. In addition, the effect of Matrine on caspase-3 activation was investigated by Western blot. Indirect immunofluorescence assay and Western blot analysis demonstrated that Matrine could inhibit N protein expression in Marc-145 cells. And Matrine was found to be able to impair PRRSV-induced apoptosis by inhibiting caspase-3 activation.

  17. Prioritising weed management activities in a data deficient environment: the Pilbara islands, Western Australia.

    PubMed

    Lohr, Cheryl; Passeretto, Kellie; Lohr, Michael; Keighery, Greg

    2015-12-01

    Along the Pilbara coast of Western Australia (WA) there are approximately 598 islands with a total area of around 500 km(2). Budget limitations and logistical complexities mean the management of these islands tends to be opportunistic. Until now there has been no review of the establishment and impacts of weeds on Pilbara islands or any attempt to prioritise island weed management. In many instances only weed occurrence has been documented, creating a data deficient environment for management decision making. The purpose of this research was to develop a database of weed occurrences on WA islands and to create a prioritisation process that will generate a ranked list of island-weed combinations using currently available data. Here, we describe a model using the pairwise comparison formulae in the Analytical Hierarchy Process (AHP), four metrics describing the logistical difficulty of working on each island (island size, ruggedness, travel time, and tenure), and two well established measures of conservation value of an island (maximum representation and effective maximum rarity of eight features). We present the sensitivity of the island-weed rankings to changes in weights applied to each decision criteria using Kendall's tau statistics. We also present the top 20 ranked island-weed combinations for four modelling scenarios. Many conservation prioritisation tools exist. However, many of these tools require extrapolation to fill data gaps and require specific management objectives and dedicated budgets. To our knowledge, this study is one of a few attempts to prioritise conservation actions using data that are currently available in an environment where management may be opportunistic and spasmodic due to budgetary restrictions. PMID:27441230

  18. Prioritising weed management activities in a data deficient environment: the Pilbara islands, Western Australia.

    PubMed

    Lohr, Cheryl; Passeretto, Kellie; Lohr, Michael; Keighery, Greg

    2015-12-01

    Along the Pilbara coast of Western Australia (WA) there are approximately 598 islands with a total area of around 500 km(2). Budget limitations and logistical complexities mean the management of these islands tends to be opportunistic. Until now there has been no review of the establishment and impacts of weeds on Pilbara islands or any attempt to prioritise island weed management. In many instances only weed occurrence has been documented, creating a data deficient environment for management decision making. The purpose of this research was to develop a database of weed occurrences on WA islands and to create a prioritisation process that will generate a ranked list of island-weed combinations using currently available data. Here, we describe a model using the pairwise comparison formulae in the Analytical Hierarchy Process (AHP), four metrics describing the logistical difficulty of working on each island (island size, ruggedness, travel time, and tenure), and two well established measures of conservation value of an island (maximum representation and effective maximum rarity of eight features). We present the sensitivity of the island-weed rankings to changes in weights applied to each decision criteria using Kendall's tau statistics. We also present the top 20 ranked island-weed combinations for four modelling scenarios. Many conservation prioritisation tools exist. However, many of these tools require extrapolation to fill data gaps and require specific management objectives and dedicated budgets. To our knowledge, this study is one of a few attempts to prioritise conservation actions using data that are currently available in an environment where management may be opportunistic and spasmodic due to budgetary restrictions.

  19. Western Gas Sands Project. Quarterly basin activities report, January 1-March 31, 1980

    SciTech Connect

    Not Available

    1980-01-01

    This report is a summary of drilling and testing activities in the four primary study areas of the WGSP: Greater Green River Basin, Northern Great Plains Province, Uinta Basin, and Piceance Basin. (DLC)

  20. [Effects of snow pack on soil nitrogen transformation enzyme activities in a subalpine Abies faxioniana forest of western Sichuan, China].

    PubMed

    Xiong, Li; Xu, Zhen-Feng; Wu, Fu-Zhong; Yang, Wan-Qin; Yin, Rui; Li, Zhi-Ping; Gou, Xiao-Lin; Tang, Shi-Shan

    2014-05-01

    This study characterized the dynamics of the activities of urease, nitrate reductase and nitrite reductase in both soil organic layer and mineral soil layer under three depths of snow pack (deep snowpack, moderate snowpack and shallow snowpack) over the three critical periods (snow formed period, snow stable period, and snow melt period) in the subalpine Abies faxoniana forest of western Sichuan in the winter of 2012 and 2013. Throughout the winter, soil temperature under deep snowpack increased by 46.2% and 26.2%, respectively in comparison with moderate snowpack and shallow snowpack. In general, the three nitrogen-related soil enzyme activities under shallow snowpack were 0.8 to 3.9 times of those under deep snowpack during the winter. In the beginning and thawing periods of seasonal snow pack, shallow snowpack significantly increased the activities of urease, nitrate and nitrite reductase enzyme in both soil organic layer and mineral soil layer. Although the activities of the studied enzymes in soil organic layer and mineral soil layer were observed to be higher than those under deep- and moderate snowpacks in deep winter, no significant difference was found under the three snow packs. Meanwhile, the effects of snowpack on the activities of the measured enzymes were related with season, soil layer and enzyme type. Significant variations of the activities of nitrogen-related enzymes were found in three critical periods over the winter, and the three measured soil enzymes were significantly higher in organic layer than in mineral layer. In addition, the activities of the three measured soil enzymes were closely related with temperature and moisture in soils. In conclusion, the decrease of snow pack induced by winter warming might increase the activities of soil enzymes related with nitrogen transformation and further stimulate the process of wintertime nitrogen transformation in soils of the subalpine forest.

  1. 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

  2. Installation of a digital, wireless, strong-motion network for monitoring seismic activity in a western Colorado coal mining region

    SciTech Connect

    Peter Swanson; Collin Stewart; Wendell Koontz

    2007-01-15

    A seismic monitoring network has recently been installed in the North Fork Valley coal mining region of western Colorado as part of a NIOSH mine safety technology transfer project with two longwall coal mine operators. Data recorded with this network will be used to characterize mining related and natural seismic activity in the vicinity of the mines and examine potential hazards due to ground shaking near critical structures such as impoundment dams, reservoirs, and steep slopes. Ten triaxial strong-motion accelerometers have been installed on the surface to form the core of a network that covers approximately 250 square kilometers (100 sq. miles) of rugged canyon-mesa terrain. Spread-spectrum radio networks are used to telemeter continuous streams of seismic waveform data to a central location where they are converted to IP data streams and ported to the Internet for processing, archiving, and analysis. 4 refs.

  3. Bat activity in thinned, unthinned, and old-growth forests in western Oregon

    USGS Publications Warehouse

    Humes, Marcia L.; Hayes, J.P.; Collopy, M.W.

    1999-01-01

    Many aspects of the influences of forest management activities on bats (Chiroptera) in the Pacific Northwest are poorly known. We compared thinned and unthinned forest stands of the same age and old-growth forest stands to determine potential differences in structure and amount of use by bats. We hypothesized that activity levels of bats would differ in stands differing in structure as a result of management history and that activity of bats would be similar in stands of similar structure. We used automated ultrasonic detectors (Anabat II) to record calls of bats in 50-100-year-old thinned and unthinned stands, and in old-growth (a?Y200 yr old) stands in the Oregon Coast Range during the summers of 1994 and 1995. Our median index of bat activity was higher in old-growth than in unthinned stands and higher in thinned than in unthinned stands. We were not able to detect a significant difference between the index of median bat activity for old-growth and thinned stands. More than 90% of identifiable passes were identified as calls from Myotis species. The 3 stand types we examined differed in certain structural characteristics such as density and size of trees, and amount of overstory and understory cover. We concluded that the structural changes caused by thinning may benefit bats by creating habitat structure in young stands that bats are able to use more effectively.

  4. Morphological changes in the blastocyst of the western spotted skunk during activation from delayed implantation.

    PubMed

    Enders, A C; Schlafke, S; Hubbard, N E; Mead, R A

    1986-03-01

    Blastocysts collected from the spotted skunk during delay of implantation, early activation and late activation demonstrate three-tiered growth and developmental changes. The slow-growing blastocyst from the several months of delay is small (less than 1.1 mm) with a rounded inner cell mass consisting of clusters of rounded, lipid-filled cells. During the several days of early activation, the lipid in both inner cell mass and trophoblast diminishes, polyribosomes increase in number, and the endodermal layer differentiates as the blastocyst grows (1.2-1.6 mm). At activation the inner cell mass flattens, becomes uncovered by polar trophoblast, and forms a disc of columnar epiblast cells. The blastocyst expands rapidly during the last 24-48 h prior to implantation to 1.7-2.0 mm, and the trophoblast becomes cuboidal with a marked endocytotic apparatus. The morphological evidence, together with previous studies of protein and RNA synthesis, suggests a tooling-up period during early activation with progressive increases in rates of growth and differentiation in the last hours as implantation approaches.

  5. [L-Lysine-α-Oxidase in vitro Activity in Experiments on Models of Viruses Sindbis, Forest-Spring Encephalitis, Western Nile, Tyaginya and Dhori].

    PubMed

    Smirnova, I P; Larichev, V F; Shneider, Yu A

    2015-01-01

    The antitumor effect of L-lysine-α-oxidase from the culture fluid of Trichoderma harzianum Rifai F-180 was investigated for the first time. The in vitro studies revealed its high activity on a model of the forest-spring encephalitis virus and no activity against the Sindbis, Western Nile, Tyaginya and Dhori viruses. PMID:26415376

  6. Differential methane oxidation activity and microbial community composition at cold seeps in the Arctic off western Svalbard

    NASA Astrophysics Data System (ADS)

    Gründger, Friederike; Svenning, Mette M.; Niemann, Helge; Silyakova, Anna; Serov, Pavel; Li Hong, Wei; Wegener, Gunter; Panieri, Giuliana; Carroll, JoLynn

    2016-04-01

    Most models considering climate change related bottom water warming suggest that gas hydrates may become destabilized, leading to the mobilization of methane into seabed and water column ecosystems, and, eventually, into the atmosphere. However, the capacity of methanotrophic microbes retaining methane in sediments and the hydrosphere is not well constrained. Here, we investigate the microbial utilization of methane in sediments and the water column, focusing on cold seeps discovered at the arctic continental margin of western Svalbard. We measured ex situ rates of methane oxidation and sulfate reduction in two active gas flare sites with different geological settings at the Vestnesa Ridge (1204 m water depth) and within a pingolike feature area southwest off Svalbard (PLF; 380 m water depth). Our results show contrarily situations at our two sampling sites: At Vestnesa Ridge we find high methane oxidation rates with values up to 2055 nmol cm-3 d-1 at the sediment surface where the sediments are oversaturated with methane. Whereas, methane concentration and oxidation rates are low in the overlying water column (2 pmol cm-3 d-1). In contrast, at the sediment surface at PLF methane concentration and oxidation rates are considerably lower (up to 1.8 nmol cm-3 d-1). While the overlying bottom water contains high concentration of methane and shows oxidation rates with values of up to 3.8 nmol cm-3 d-1. The data on methane oxidation and sulfate reduction activity are compared to the sediment geochemistry and to data from metagenomic analysis identifying the methanotrophic community composition. These results provide unique insight into the dynamic responses of the seabed biological filter at cold seeps in the Arctic off western Svalbard. This study is part of the Centre for Arctic Gas Hydrate, Environment and Climate and was supported by the Research Council of Norway through its Centres of Excellence funding scheme grant No. 223259.

  7. Ecological Activism in Post-Soviet Russia and the Western World (A Comparative Analysis)

    ERIC Educational Resources Information Center

    Usacheva, O. A.

    2012-01-01

    Ecological activism (henceforth ecoactivism) in Russia, a country with a predominant European culture, has common roots with the Europe of the late 19th and early 20th centuries. A process of intensive industrialization and urbanization required that unspoiled, natural landscapes be preserved for rest, recreation, and ecological education. This…

  8. Searching for Active Faults in the Western Eurasia-Nubia plate boundary

    NASA Astrophysics Data System (ADS)

    Antunes, Veronica; Custodio, Susana; Arroucau, Pierre; Carrilho, Fernando

    2016-04-01

    The repeated occurrence of large magnitude earthquakes in southwest Iberia in historical and instrumental times suggests the presence of active faults in the region. However, the region undergoes slow deformation, which results in low rates of seismic activity, and the location, dimension and geometry of active structures remains unsettled. We recently developed a new algorithm for earthquake location in 3D complex media with laterally varying interface depths, which allowed us to relocate 2363 events that occurred from 2007 to 2013. The method takes as inputs P- and S-wave catalog arrival times obtained from the Portuguese Meteorological Institute (IPMA, Instituto Portugues do Mar e da Atmosfera), for a study area defined by 8.5°W < lon < 5°W and 36° < lat < 37.5°. After relocation, we obtain a lineation of events in the Guadalquivir bank region, in the northern Gulf of Cadiz. The lineation defines a low-angle northward-dipping plane rooted at the base of the crust, which could indicate the presence of a major fault. We provide seismological evidence for the existence of this seemingly active structure based on earthquake relocations, focal mechanisms and waveform similarity between neighboring events.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Aims: To compare the ability of the Italian and the U.S. bovine spongiform encephalopathy (BSE) confirmatory protocols in the detection of classical (C-) and atypical - low (L-) and high (H-) type- BSE forms. Methods and Results: Obex samples from U.S. and Italian C-type BSE cases, a U.S. H-type and...

  10. Identification of hantavirus infection by Western blot assay and TaqMan PCR in patients hospitalized with acute kidney injury.

    PubMed

    Oldal, Miklós; Németh, Viktória; Madai, Mónika; Kemenesi, Gábor; Dallos, Bianka; Péterfi, Zoltán; Sebők, Judit; Wittmann, István; Bányai, Krisztián; Jakab, Ferenc

    2014-06-01

    Hantaviruses, one of the causative agents of viral hemorrhagic fevers, represent a considerable healthcare threat. In Hungary, Dobrava-Belgrade virus (DOBV) and Puumala virus (PUUV) are the main circulating hantavirus species, responsible for the clinical picture known as hemorrhagic fever with renal syndrome, a disease that may be accompanied by acute kidney injury (AKI), requiring hospitalization with occasionally prolonged recovery phase. A total of 20 patient sera were collected over a 2-year period from persons hospitalized with AKI, displaying clinical signs and laboratory findings directly suggestive for hantavirus infection. Samples were tested using an immunoblot assay, based on complete viral nucleocapsid proteins to detect patients' IgM and IgG antibodies against DOBV and PUUV. In parallel, all specimens were also tested by 1-step real-time TaqMan reverse-transcriptase polymerase chain reaction to confirm infection and to determine the causative hantavirus genotype. We present here the first Hungarian clinical study spanning across 2 years and dedicated specifically to assess acute kidney injuries, in the context of hantavirus prevalence.

  11. Promoting Physics Among Female Learners in the Western Cape Through Active Engagement (abstract)

    NASA Astrophysics Data System (ADS)

    Arendse, Gillian J.

    2009-04-01

    In 2006 the author organized a one-day intervention aimed at promoting physics among female learners at the University of Stellenbosch. The activities included an interactive lecture demonstration promoting active engagement, a hands-on session, and short presentations by female physicists addressing issues such as balancing family and career, breaking the stereotypes, and launching a successful career in physics. Each learner was expected to evaluate the program. In 2007 the author joined forces with Hip2B2 (Shuttleworth Foundation) to host a competition among grade-10 learners with the theme, ``promoting creativity through interactivity.'' The author was tasked by the Hip2B2-team to assist with a program for female learners planned for August 2008, coinciding with our national celebration of Women's Day. The event targeted 160 learners and took place in Durban, East London, Cape Town, and Johannesburg. The author shares some of the learners' experiences and personal triumphs.

  12. Geodynamic significance of the TRM segment in the East African Rift: active tectonics and paleostress in western Tanzania

    NASA Astrophysics Data System (ADS)

    Delvaux, D.; Kervyn, F.; Macheyeki, A. S.; Temu, E. B.

    2012-04-01

    The Tanganyika-Rukwa-Malawi (TRM) rift segment in western Tanzania is a key sector for understanding the opening dynamics of the East African rift system (EARS). In an oblique opening model, it is considered as a dextral transfer fault zone that accommodates the general opening of the EARS in a NW-SE direction. In an orthogonal opening model, it accommodates pure dip-slip normal faulting with extension orthogonal to the rift segments and a general E-W extension for the entire EARS. We investigated the active tectonic architecture and paleostress evolution of the Ufipa plateau and adjacent Rukwa basin and in order to define their geodynamic role in the development of the EARS and highlight their pre-rift brittle tectonic history. The active fault architecture, fault-kinematic analysis and paleostress reconstruction show that the recent to active fault systems that control the rift structure develop in a pure extensional setting with extension direction orthogonal to the trend of the TRM segment. Two pre-rift brittle events are evidenced. An older brittle thrusting is related to the interaction between the Bangweulu block and the Tanzanian craton during the late Pan-African (early Paleozoic). It was followed by a transpressional inversion during the early Mesozoic. This inversion stage caused dextral strike-slip faulting along the fault systems that now control the major rift structures. It has been erroneously interpreted as related to the late Cenozoic EARS which instead is characterized by pure normal faulting.

  13. Middle Miocene hiatus in volcanic activity in the Great Basin area of the Western United States

    USGS Publications Warehouse

    McKee, E.H.; Noble, D.C.; Silberman, M.L.

    1970-01-01

    A summary of potassium-argon dates shows that a high level of igneous activity in the Great Basin and adjacent regions during middle Tertiary time (40 to 20 my ago) was followed by a period of relative quiescence in middle Miocene time that lasted for several million years (from 20 to 17 my ago). Volcanism resumed 16 my ago mainly at the margins of the region and has continued to the present. ?? 1970.

  14. Normal faulting along the western side of the Matese Mountains: Implications for active tectonics in the Central Apennines (Italy)

    NASA Astrophysics Data System (ADS)

    Boncio, Paolo; Dichiarante, Anna Maria; Auciello, Eugenio; Saroli, Michele; Stoppa, Francesco

    2016-01-01

    We provide new field data from geologic mapping and bedrock structural geology along the western side of the Matese Mts in central Italy, a region of high seismicity, strain rates among the highest of the entire Apennines (4-5 mm/yr GPS-determined extension), and poorly constrained active faults. The existing knowledge on the Aquae Iuliae normal fault (AIF) was implemented with geometric and kinematic data that better constrain its total length (16.5 km), the minimum long-term throw rate (0.3-0.4 mm/yr, post-late glacial maximum, LGM), and the segmentation. For the first time, we provide evidence of post-350 ka and possibly late Quaternary activity of the Ailano - Piedimonte Matese normal fault (APMF). The APMF is 18 km long. It is composed of a main 11 km-long segment striking NW-SE and progressively bending to the E-W in its southern part, and a 7 km-long segment striking E-W to ENE-WSW with very poor evidence of recent activity. The available data suggest a possible post-LGM throw rate of the main segment of ≳0.15 mm/yr. There is no evidence of active linkage in the step-over zone between the AIF and APMF (Prata Sannita step-over). An original tectonic model is proposed by comparing structural and geodetic data. The AIF and APMF belong to two major, nearly parallel fault systems. One system runs at the core of the Matese Mts and is formed by the AIF and the faults of the Gallo-Letino-Matese Lake system. The other system runs along the western side of the Matese Mts and is formed by the APMF, linked to the SE with the Piedimonte Matese - Gioia Sannitica fault. The finite extension of the APMF might be transferred to the NW towards the San Pietro Infine fault. The nearly 2-3 mm/yr GPS-determined extension rate is probably partitioned between the two systems, with a ratio that is difficult to establish due to poor GPS coverage. The proposed model, though incomplete (several faults/transfer zones need further investigations), aids in the seismotectonic

  15. Novel Indole-2-Carboxamide Compounds Are Potent Broad-Spectrum Antivirals Active against Western Equine Encephalitis Virus In Vivo

    PubMed Central

    Delekta, Phillip C.; Dobry, Craig J.; Sindac, Janice A.; Barraza, Scott J.; Blakely, Pennelope K.; Xiang, Jianming; Kirchhoff, Paul D.; Keep, Richard F.; Irani, David N.; Larsen, Scott D.

    2014-01-01

    ABSTRACT Neurotropic alphaviruses, including western, eastern, and Venezuelan equine encephalitis viruses, cause serious and potentially fatal central nervous system infections in humans for which no currently approved therapies exist. We previously identified a series of thieno[3,2-b]pyrrole derivatives as novel inhibitors of neurotropic alphavirus replication, using a cell-based phenotypic assay (W. Peng et al., J. Infect. Dis. 199:950–957, 2009, doi:http://dx.doi.org/10.1086/597275), and subsequently developed second- and third-generation indole-2-carboxamide derivatives with improved potency, solubility, and metabolic stability (J. A. Sindac et al., J. Med. Chem. 55:3535–3545, 2012, doi:http://dx.doi.org/10.1021/jm300214e; J. A. Sindac et al., J. Med. Chem. 56:9222–9241, 2013, http://dx.doi.org/10.1021/jm401330r). In this report, we describe the antiviral activity of the most promising third-generation lead compound, CCG205432, and closely related analogs CCG206381 and CCG209023. These compounds have half-maximal inhibitory concentrations of ∼1 μM and selectivity indices of >100 in cell-based assays using western equine encephalitis virus replicons. Furthermore, CCG205432 retains similar potency against fully infectious virus in cultured human neuronal cells. These compounds show broad inhibitory activity against a range of RNA viruses in culture, including members of the Togaviridae, Bunyaviridae, Picornaviridae, and Paramyxoviridae families. Although their exact molecular target remains unknown, mechanism-of-action studies reveal that these novel indole-based compounds target a host factor that modulates cap-dependent translation. Finally, we demonstrate that both CCG205432 and CCG209023 dampen clinical disease severity and enhance survival of mice given a lethal western equine encephalitis virus challenge. These studies demonstrate that indole-2-carboxamide compounds are viable candidates for continued preclinical development as inhibitors of

  16. Novel indole-2-carboxamide compounds are potent broad-spectrum antivirals active against western equine encephalitis virus in vivo.

    PubMed

    Delekta, Phillip C; Dobry, Craig J; Sindac, Janice A; Barraza, Scott J; Blakely, Pennelope K; Xiang, Jianming; Kirchhoff, Paul D; Keep, Richard F; Irani, David N; Larsen, Scott D; Miller, David J

    2014-10-01

    Neurotropic alphaviruses, including western, eastern, and Venezuelan equine encephalitis viruses, cause serious and potentially fatal central nervous system infections in humans for which no currently approved therapies exist. We previously identified a series of thieno[3,2-b]pyrrole derivatives as novel inhibitors of neurotropic alphavirus replication, using a cell-based phenotypic assay (W. Peng et al., J. Infect. Dis. 199:950-957, 2009, doi:http://dx.doi.org/10.1086/597275), and subsequently developed second- and third-generation indole-2-carboxamide derivatives with improved potency, solubility, and metabolic stability (J. A. Sindac et al., J. Med. Chem. 55:3535-3545, 2012, doi:http://dx.doi.org/10.1021/jm300214e; J. A. Sindac et al., J. Med. Chem. 56:9222-9241, 2013, http://dx.doi.org/10.1021/jm401330r). In this report, we describe the antiviral activity of the most promising third-generation lead compound, CCG205432, and closely related analogs CCG206381 and CCG209023. These compounds have half-maximal inhibitory concentrations of ∼1 μM and selectivity indices of >100 in cell-based assays using western equine encephalitis virus replicons. Furthermore, CCG205432 retains similar potency against fully infectious virus in cultured human neuronal cells. These compounds show broad inhibitory activity against a range of RNA viruses in culture, including members of the Togaviridae, Bunyaviridae, Picornaviridae, and Paramyxoviridae families. Although their exact molecular target remains unknown, mechanism-of-action studies reveal that these novel indole-based compounds target a host factor that modulates cap-dependent translation. Finally, we demonstrate that both CCG205432 and CCG209023 dampen clinical disease severity and enhance survival of mice given a lethal western equine encephalitis virus challenge. These studies demonstrate that indole-2-carboxamide compounds are viable candidates for continued preclinical development as inhibitors of neurotropic

  17. Structural Evidence for Fault Reactivation: the Active Priene-Sazli Fault Zone, Söke-Milet Basin, Western Anatolia

    NASA Astrophysics Data System (ADS)

    Sümer, Ö.; Inci, U.; Sözbilir, H.; Uzel, B.

    2009-04-01

    Western Anatolia is located at tha eastern part of the Aegean region that forms one of the most seismically active and rapidly extending regions in the world. One of the most prominent structural component of the Western Anatolia is E-W trending grabens. One of them is the Büyük Menderes Graben (BMG) showing a major change in strike ranging from E-W to NE-SW in its western end. This NE-SW oriented part of the graben is known as the Söke-Milet basin (SMB). The depression is 35 km long and 16 km wide. NW border of the basin is characterized by a morphotectonic structure namely Priene-Sazlı fault zone (PSFZ). The 16 July 1955 Söke-Balat earthquake (M=6.8) was atributed to this fault (Eyidogan and Jackson, 1985; Sengör, 1987; Altunel, 1998). However, field based kinematic studies on the PSFZ are lacking except for Gürer et. al. (2001). In this paper, we studied several reactivated fault segments of the PSFZ that are repeatedly formed under changing stress fields in order to evaluate the kinematic and stress history of the region by using structural relationships between striations and fault-plane related structures. The PSFZ consists of 5 fault segments which are en échelon arranged on the basis of mapping geological structures. The northern segments that strikes NE in the north and bends into an approximately E-W direction around Doganbey to the SW. Each segment is identified as steep opographic scarps ranging in height from a few meters to several hundred meters. Fault segments become to linkage and show breaching of the relay ramps between them. We interpret that such fault patterns have been formed in a region where extension has reactivated on pre-existing structures in an oblique sense. Evidence for this is the presence of three sets of striations each with different orientations on the same slip surface of the studied fault segments. Here, two differently oriented strike-slip slickenlines are postdated by dip-slip striations. Based on our structural

  18. Analysis of methanogenic and methanotrophic activity at the western margin of the Greenland Ice Sheet

    NASA Astrophysics Data System (ADS)

    Broemsen, E. L.; Webster, K. D.; Dieser, M.; Pratt, L. M.; Christner, B. C.

    2012-12-01

    Anoxic conditions in environments beneath the world's glaciers and ice sheets provide plausible habitats supporting the microbial production of methane. Recent reports of potential methane sources beneath the Greenland Ice Sheet (GrIS) suggest in situ production by an active community of methanogens. Beneath the GrIS, microbially derived methane can be dissolved in subglacial water, and during periods of melting, can exchange with the atmosphere at sites of subglacial discharge. Transfer of methane from subglacial fluids to the atmosphere could be a significant climate factor, but few data are available to make such assessments. The specific aim of this study was to characterize the composition and activity of methanogens and methanotrophs present in samples of subglacial outflow at the ice sheet margin near Kangerlussuaq, Greenland. Subglaical water was collected twice-weekly over a nine week period (mid July to mid September of 2012) and the dissolved methane concentration in the samples was determined via gas chromatography. Extracted RNA and DNA from the subglacial water was analyzed by analysis of 16s rRNA and rRNA genes present in the subglacial assemblages. From the molecular results we infer the presence of active methanogens related to the order Methanosarcinales. Further, locally elevated concentrations of atmospheric methane as high as 1.92 ± 0.03 ppmv, were detected in the ice tunnel of the subglacial outflow using open-path laser spectrometry. From these data we estimate rates of methane release at the ice sheet margin during the summer melt months at this geographical location. The results provide a context for addressing the impact that deglaciation will have on the release of greenhouse gases from ice sheets on a warming Earth.

  19. Active transtensional intracontinental basins: Walker Lane in the western Great Basin

    USGS Publications Warehouse

    Jayko, Angela S.; Bursik, Marcus

    2012-01-01

    The geometry and dimensions of sedimentary basins within the Walker Lane are a result of Plio-Pleistocene transtensive deformation and partial detachment of the Sierra Nevada crustal block from the North American plate. Distinct morpho-tectonic domains lie within this active transtensive zone. The northeast end of the Walker Lane is partly buried by active volcanism of the southern Cascades, and adjacent basins are filled or poorly developed. To the south, the basin sizes are moderate, 25–45km × 15–10 km, with narrow 8-12km wide mountain ranges mainly oriented N-S to NNE. These basins form subparallel arrays in discrete zones trending about 300° and have documented clockwise rotation. This is succeeded to the south by a releasing stepover domain ∼85-100km wide, where the basins are elongated E-W to ENE, small (∼15-30km long, 5-15km wide), and locally occupied by active volcanic centers. The southernmost part of the Walker Lane is structurally integrated, with high to extreme relief. Adjacent basins are elongate, 50-200km long and ∼5 -20km wide. Variations in transtensive basin orientations in the Walker Lane are largely attributable to variations in strain partitioning. Large basins in the Walker Lane have 2-6km displacement across basin bounding faults with up to 3 km of clastic accumulation based on gravity and drill hole data. The sedimentary deposits of the basins may include interbedded volcanic deposits with bimodal basaltic and rhyolitic associations. The basins may include lacustrine deposits that record a wide range of water chemistry from cold fresh water conditions to saline-evaporative

  20. Evolution of magmatic and hydrothermal activity in the western Arctic and North Atlantic regions

    NASA Astrophysics Data System (ADS)

    Sorokhtin, N. O.; Lobkovsky, L. I.; Novikov, G. V.; Kozlov, E. E.; Bogdanova, O. Yu.; Nikiforov, S. L.

    2016-07-01

    This paper discusses the geodynamic evolution of the lithosphere in the Arctic region during the Phaneorozic and its polyphase lithotectonic reorganization. Spatiotemporal patterns of the mosaic junction of lithospheric plates of different age are presented for the Caledonian-Hercynian stage and for the Cenozoic evolution of the North Atlantic and Arctic oceanic basins. Special attention is given to the intersections of fault systems with different kinematics, which control the manifestation of peculiar magmatism and the formation of numerous mineral deposits. It is shown that the hydrothermal activity of the region is related to the ocean opening in the Eocene and is confined to the mid-ocean ridge.

  1. Active low-angle (?) normal faulting along the North Lunggar rift, western Tibet

    NASA Astrophysics Data System (ADS)

    Logan, M. A.; Taylor, M. H.; Styron, R. H.; Gosse, J. C.; Ding, L.; Yang, G.

    2012-12-01

    Here we present surface exposure ages of faulted fluvial terraces using cosmogenic nuclides from the North Lunggar rift. The Lunggar rift is one of seven major north-striking rift basins accommodating east-west directed extension on the Tibetan Plateau. The Lunggar rift in west-central Tibet is divided into two distinct north and south segments based on fault geometry. The North Lunggar range is bounded on its east side by a <40 degree dipping, ~N-striking normal fault. This normal fault is considered inactive as the main detachment is unconformably overlain by unfaulted moraines and alluvial fans. Farther into the hanging wall basin, approximately 6 km eastward, several fault scarps parallel the Lunggar detachment. Locally, active faulting is distributed in the hanging wall with as many as seven normal fault scarps accommodating active east-west directed extension. Recent activity of these smaller faults is apparent from cross-cut fluvial terraces that have been uplifted by as much as 75 m. The geomorphology and fault geometry of the North Lunggar rift are consistent with high-angle normal faults that sole into a single master detachment fault at depth. A high-resolution digital elevation model constructed from real-time kinematic-GPS data has made details of the geomorphology clear and allowed for precise measurements of geomorphic offsets across the fault scarps. We estimate the surface abandonment ages using the depth profiling approach with cosmogenic nuclides. Three cosmogenic depth profiles are being analyzed in this study with each depth profile consisting of five samples at varying depths in order to account for inheritance. Site 1 is the southernmost and is on the highest uplifted fluvial terrace and is being prepared for 10Be analysis. Site 2 comprises two depth profiles on the highest and intermediate uplifted terraces, respectively. Samples at site 2 have low quartz yields and are being prepared for 36Cl analysis. Combining the fault offsets and

  2. Ensemble projections of wildfire activity and carbonaceous aerosol concentrations over the western United States in the mid-21st century

    NASA Astrophysics Data System (ADS)

    Yue, Xu; Mickley, Loretta J.; Logan, Jennifer A.; Kaplan, Jed O.

    2013-10-01

    We estimate future wildfire activity over the western United States during the mid-21st century (2046-2065), based on results from 15 climate models following the A1B scenario. We develop fire prediction models by regressing meteorological variables from the current and previous years together with fire indexes onto observed regional area burned. The regressions explain 0.25-0.60 of the variance in observed annual area burned during 1980-2004, depending on the ecoregion. We also parameterize daily area burned with temperature, precipitation, and relative humidity. This approach explains ˜0.5 of the variance in observed area burned over forest ecoregions but shows no predictive capability in the semi-arid regions of Nevada and California. By applying the meteorological fields from 15 climate models to our fire prediction models, we quantify the robustness of our wildfire projections at midcentury. We calculate increases of 24-124% in area burned using regressions and 63-169% with the parameterization. Our projections are most robust in the southwestern desert, where all GCMs predict significant (p < 0.05) meteorological changes. For forested ecoregions, more GCMs predict significant increases in future area burned with the parameterization than with the regressions, because the latter approach is sensitive to hydrological variables that show large inter-model variability in the climate projections. The parameterization predicts that the fire season lengthens by 23 days in the warmer and drier climate at midcentury. Using a chemical transport model, we find that wildfire emissions will increase summertime surface organic carbon aerosol over the western United States by 46-70% and black carbon by 20-27% at midcentury, relative to the present day. The pollution is most enhanced during extreme episodes: above the 84th percentile of concentrations, OC increases by ˜90% and BC by ˜50%, while visibility decreases from 130 km to 100 km in 32 Federal Class 1 areas in

  3. Ensemble projections of wildfire activity and carbonaceous aerosol concentrations over the western United States in the mid-21st century.

    PubMed

    Yue, Xu; Mickley, Loretta J; Logan, Jennifer A; Kaplan, Jed O

    2013-10-01

    We estimate future wildfire activity over the western United States during the mid-21(st) century (2046-2065), based on results from 15 climate models following the A1B scenario. We develop fire prediction models by regressing meteorological variables from the current and previous years together with fire indexes onto observed regional area burned. The regressions explain 0.25-0.60 of the variance in observed annual area burned during 1980-2004, depending on the ecoregion. We also parameterize daily area burned with temperature, precipitation, and relative humidity. This approach explains ~0.5 of the variance in observed area burned over forest ecoregions but shows no predictive capability in the semi-arid regions of Nevada and California. By applying the meteorological fields from 15 climate models to our fire prediction models, we quantify the robustness of our wildfire projections at mid-century. We calculate increases of 24-124% in area burned using regressions and 63-169% with the parameterization. Our projections are most robust in the southwestern desert, where all GCMs predict significant (p<0.05) meteorological changes. For forested ecoregions, more GCMs predict significant increases in future area burned with the parameterization than with the regressions, because the latter approach is sensitive to hydrological variables that show large inter-model variability in the climate projections. The parameterization predicts that the fire season lengthens by 23 days in the warmer and drier climate at mid-century. Using a chemical transport model, we find that wildfire emissions will increase summertime surface organic carbon aerosol over the western United States by 46-70% and black carbon by 20-27% at midcentury, relative to the present day. The pollution is most enhanced during extreme episodes: above the 84(th) percentile of concentrations, OC increases by ~90% and BC by ~50%, while visibility decreases from 130 km to 100 km in 32 Federal Class 1 areas in

  4. Zooplankton biomass and electron transport system activity around the Balearic Islands (western Mediterranean)

    NASA Astrophysics Data System (ADS)

    Herrera, A.; Gómez, M.; Packard, T. T.; Fernández de Puelles, M. L.

    2014-03-01

    Measuring electron transport system (ETS) activity in zooplankton provides an index of respiration, theoretically, the potential respiration rate. We apply the ETS technique to estimate potential respiration and carbon demand from the zooplankton community in the upper 200 m of the water column near the Balearic Islands. The investigation was focused on two areas with different oceanographic conditions: the Balearic and Algerian subbasins. It compared the biomass, potential respiration and specific potential respiration of different size fractions (53-200, 200-500, > 500 μm) in both areas. In these regions the largest contribution to respiration was found in the larger sizes. The specific respiration (per unit biomass) was greater in smaller fractions, indicating that they have a more active metabolism. Both biomass and potential respiration increased in the Algerian subbasin and for both regions biomass and potential respiration were greater in shallow waters over the continental shelf (< 200 m). Using Kleiber's law as a tool to investigate the relationships between these two variables, we found that the exponential relation coefficient (b) was less than 0.75, indicating that the respiration was depressed (shifted down). In cultures and in eutrophic ocean waters (upwelling areas) b normally is greater than 0.75, consequently we intuit that the low value of b over the Balearic and Algerian subbasins indicates that the zooplankton is not well fed and that they are living under oligotrophic stress.

  5. Overview of electromagnetic methods applied in active volcanic areas of western United States

    NASA Astrophysics Data System (ADS)

    Skokan, Catherine K.

    1993-06-01

    A better understanding of active volcanic areas in the United States through electromagnetic geophysical studies received foundation from the many surveys done for geothermal exploration in the 1970's. Investigations by governmental, industrial, and academic agencies include (but are not limited to) mapping of the Cascades. Long Valley/Mono area, the Jemez volcanic field, Yellowstone Park, and an area in Colorado. For one example — Mt. Konocti in the Mayacamas Mountains, California — gravity, magnetic, and seismic, as well as electromagnetic methods have all been used in an attempt to gain a better understanding of the subsurface structure. In each of these volcanic regions, anomalous zones were mapped. When conductive, these anomalies were interpreted to be correlated with hydrothermal activity and not to represent a magma chamber. Electrical and electromagnetic geophysical methods can offer valuable information in the understanding of volcanoes by being the method which is most sensitive to change in temperature and, therefore, can best map heat budget and hydrological character to aid in prediction of eruptions.

  6. Dramatic increase in SHP2 binding activity of Helicobacter pylori Western CagA by EPIYA-C duplication: its implications in gastric carcinogenesis

    PubMed Central

    Nagase, Lisa; Hayashi, Takeru; Senda, Toshiya; Hatakeyama, Masanori

    2015-01-01

    Infection with cagA-positive Helicobacter pylori is critically associated with the development of gastric cancer. The cagA-encoded CagA is delivered into gastric epithelial cells via type IV secretion, where it interacts with and thereby deregulates the pro-oncogenic phosphatase SHP2. East Asian CagA and Western CagA are two major CagA species produced by H. pylori circulating in East Asian countries and in the rest of the world, respectively. The SHP2 binding site of Western CagA, termed the EPIYA-C segment, variably duplicates and infection with H. pylori carrying Western CagA with multiple EPIYA-C segments is a distinct risk factor of gastric cancer. Here we show that duplication of EPIYA-C from one to two or more increases SHP2 binding of Western CagA by more than one hundredfold. Based on the decisive difference in SHP2 binding, Western CagA can be divided into two types: type I CagA carrying a single EPIYA-C segment and type II CagA carrying multiple EPIYA-C segments. Gastric epithelial cells expressing type II CagA acquire the ability to invade extracellular matrices, a malignant cellular trait associated with deregulated SHP2. A big leap in SHP2 binding activity may therefore provide molecular basis that makes type II Western CagA a distinct gastric cancer risk. PMID:26507409

  7. 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.

  8. 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.

  9. 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

  10. Binary toxins from Bacillus thuringiensis active against the western corn rootworm, Diabrotica virgifera virgifera LeConte.

    PubMed

    Baum, James A; Chu, Chi-Rei; Rupar, Mark; Brown, Gregory R; Donovan, William P; Huesing, Joseph E; Ilagan, Oliver; Malvar, Thomas M; Pleau, Michael; Walters, Matthew; Vaughn, Ty

    2004-08-01

    The western corn rootworm, Diabrotica virgifera virgifera LeConte, is a significant pest of corn in the United States. The development of transgenic corn hybrids resistant to rootworm feeding damage depends on the identification of genes encoding insecticidal proteins toxic to rootworm larvae. In this study, a bioassay screen was used to identify several isolates of the bacterium Bacillus thuringiensis active against rootworm. These bacterial isolates each produce distinct crystal proteins with approximate molecular masses of 13 to 15 kDa and 44 kDa. Insect bioassays demonstrated that both protein classes are required for insecticidal activity against this rootworm species. The genes encoding these proteins are organized in apparent operons and are associated with other genes encoding crystal proteins of unknown function. The antirootworm proteins produced by B. thuringiensis strains EG5899 and EG9444 closely resemble previously described crystal proteins of the Cry34A and Cry35A classes. The antirootworm proteins produced by strain EG4851, designated Cry34Ba1 and Cry35Ba1, represent a new binary toxin. Genes encoding these proteins could become an important component of a sustainable resistance management strategy against this insect pest.

  11. Relationship between behavior, adrenal activity, and environment in zoo-housed western lowland gorillas (Gorilla gorilla gorilla).

    PubMed

    Clark, Fay E; Fitzpatrick, Malcolm; Hartley, Andy; King, Andrew J; Lee, Tracey; Routh, Andrew; Walker, Susan L; George, Katherine

    2012-01-01

    Monitoring adrenal activity through noninvasive fecal hormone sampling is rapidly gaining popularity as a tool to assess zoo animal welfare. However, few studies have sought to investigate the interrelationships between behavior, adrenal activity, and environment, and ask whether both behavioral and adrenal monitoring strategies are required to assess welfare sufficiently. We present the findings of a 9-month study of a small group (one male, two females) of Western lowland gorillas, Gorilla gorilla gorilla. First, we examined the effect of environmental variables on gorilla behavior. Second, we examined the effect of environmental variables on the concentration of fecal glucocorticoid metabolites (FGC) and the relationship between behavior and FGC. Environmental variables had similar effects on all three gorillas. Negative vigilance of visitors (NVV; staring, posturing, and charging at visitors) significantly increased in all subjects as environmental noise levels increased, and food-related behavior significantly decreased in all subjects as crowd size increased. Exhibit modifications had a number of positive effects on behavior. Notably, when privacy screens were used, NVV significantly decreased in two subjects. We found no significant effects of environmental variables on FGC. However, we did find significant relationships between behavior and FGC in one female. Specifically, her NVV was significantly higher one day before, and on the same day as, raised FGC. Also, hair plucking significantly increased in the two days following raised FGC. Overall, this study demonstrates how concurrent noninvasive fecal and behavioral monitoring can be used for gorilla welfare assessment.

  12. Fracturing and earthquake activity within the Prestahnúkur fissure swarm in the Western Volcanic Rift Zone of Iceland

    NASA Astrophysics Data System (ADS)

    Hjartardóttir, Ásta Rut; Hjaltadóttir, Sigurlaug; Einarsson, Páll; Vogfjörd, Kristín.; Muñoz-Cobo Belart, Joaquín.

    2015-12-01

    The Prestahnúkur fissure swarm is located within the ultraslowly spreading Western Volcanic Zone in Iceland. The fissure swarm is characterized by normal faults, open fractures, and evidence of subglacial fissure eruptions (tindars). In this study, fractures and faults within the Prestahnúkur fissure swarm were mapped in detail from aerial photographs to determine the extent and activity of the fissure swarm. Earthquakes during the last ~23 years were relocated to map the subsurface fault planes that they delineate. The Prestahnúkur fissure swarm is 40-80 km long and up to ~20 km wide. Most of the areas of the fissure swarm have been glacially eroded, although a part of it is covered by postglacial lava flows. The fissure swarm includes numerous faults with tens of meters vertical offset within the older glacially eroded part, whereas open fractures are found within postglacial lava flows. Comparison of relocated earthquakes and surface fractures indicates that some of the surface fractures have been activated at depth during the last ~23 years, although no dike intrusions have been ongoing. The existence of tindars nevertheless indicates that dike intrusions and rifting events do occur within the Prestahnúkur fissure swarm. The low-fracture density within postglacial lava flows and low density of postglacial eruptive fissures indicate that rifting episodes occur less often than in the faster spreading Northern Volcanic Zone.

  13. Validation of MODIS and SEVIRI Active Fire Monitoring products over Western Romania. Case study: Arad County

    NASA Astrophysics Data System (ADS)

    Oanea, Lavinia; Alina Ristea, Mihaela

    2014-05-01

    At the national level, the issue of wildfire monitoring represents a long debated topic. However, in the present situation, fire management requires various improvements in terms of detection, monitoring and post-fire analysis. The objectives of this study are to validate the data provided by MODIS (Terra and Aqua) Active Fire Monitoring and SEVIRI (MSG) FIR (Active Fire Monitoring) satellite products, with wildfires field data from The Romanian General Inspectorate for Emergency Situations (IGSU) (1), to chart the efficiency of satellite products in locating fires and study their strengths and weaknesses using a SWOT analysis (2). This is the initial step of a larger project that aims to implement an online Geographic Information System for fire management that will ease wildfire data manipulation and facilitate the decision making process. In order to do so, the current study objectives must be achieved. Our general strategy is to determine the consistency of direct (field measurements) and indirect (satellite data) observations. Depending on the amount of field information, the fire characteristics (location, frequency, extension area, moment of occurrence, type of fire, and others) will be studied through a statistical analysis. The products show some peculiar restrictiveness like spatial and temporal resolution. Specifically, we will process and interpret satellite products to identify wildfires according to the data from IGSU using specialized software. The case study for the application of these procedures is a set of fire events from Arad county - Romania, that occurred between 2007 and 2013. In order to do so, it is important to compare results from different sensors with field information through various methods and to use only consistent results. The results will play an important role in achieving the above mentioned informational system, which will integrate field information, satellite data and values of parameters that influence the evolution of

  14. Presence of an inhibitor of plasminogen activator in uterine fluid of the western spotted skunk during delayed implantation.

    PubMed

    Fazleabas, A T; Mead, R A; Rourke, A W; Roberts, R M

    1984-03-01

    This study was designed to test the hypothesis that protease inhibitors would be present in uterine fluids of the western spotted skunk during the period of delayed implantation but would decline or disappear during implantation. Blood plasma, uterine flushings and medium from endometrial cultures were collected 40-70 days preimplantation (Stage 1), 20-25 days preimplantation (Stage 2), 1-3 days preimplantation (Stage 3) and 1-5 days postimplantation (Stage 4). Protein content of the flushings was low in Stage 1 (12.9 +/- 3.0 micrograms), increased slightly (21.8 +/- 6.4 micrograms) during Stage 2 and increased markedly in Stages 3 (99 +/- 44.8 micrograms) and 4 (256 +/- 150 micrograms). No protease activity was detected in any fluid at any stage. Uterine flushings, however, contained an inhibitor of plasminogen activator (PA) of apparent molecular weight approximately 70,000. The total amount of inhibitor increased as pregnancy progressed, but specific inhibitory activity was highest during Stages 1 and 2 (1.7 and 3.2 units/micrograms protein, respectively) and declined in Stages 3 and 4 (1.2 and 0.6 units/micrograms protein, respectively). The inhibitor was present in medium from uterine cultures but was low in plasma. All of the fluids could inhibit trypsin, but not plasmin. The antitrypsin activity in uterine fluids is attributed to plasma transudate. The results demonstrate an inhibitor of PA in skunk uterine fluids but show no temporal relationship between its presence and the period of obligate delay of implantation.

  15. Evolution and dynamics of active faults in southeastern Egyptian Western Desert

    NASA Astrophysics Data System (ADS)

    Abdeen, Mamdouh

    2016-07-01

    Remote sensing data processing and analysis together with interpretation of earthquake data that are followed by extensive field studies on some of the prevailing NS and EW striking faults indicate that these faults have an intimate relationship and were formed synchronously as a conjugate Riedel shears. Parallel to the NS and the EW faults open fractures filled with blown sand dominate the area of study. The Quaternary terraces adjacent to these faults are offset by the faults. Kinematic indicators on the NS striking faults indicate major sinistral (left-lateral) strike slip and minor dip-slip (normal) movement. On the other hand, kinematic indicators on the EW striking faults indicate major dextral (right-lateral) strike slip and minor dip-slip (normal) movement. Paleo-stress analysis of the fault striae measured on the NS and EW faults indicate that these faults were formed under NNE-SSW oriented extension. Instrumental earthquake data analysis shows a comparable extension direction to that derived from field measurements of slickenlineation. These observations indicate that the NS- and EW-striking faults are contemporaneous and are related to the Red Sea rifting that is currently active.

  16. Paula Fox's "Western Wind."

    ERIC Educational Resources Information Center

    Corsaro, Julie

    1997-01-01

    Summarizes the novel, "Western Wind," by Paula Fox, lists 12 discussion questions, describes six activities, and contains an annotated bibliography of seven other novels related to birth, death, and families. (DGM)

  17. Human Activity Dampens the Benefits of Group Size on Vigilance in Khulan (Equus hemionus) in Western China.

    PubMed

    Wang, Mu-Yang; Ruckstuhl, Kathreen E; Xu, Wen-Xuan; Blank, David; Yang, Wei-Kang

    2016-01-01

    Animals receive anti-predator benefits from social behavior. As part of a group, individuals spend less time being vigilant, and vigilance decreases with increasing group size. This phenomenon, called "the many-eyes effect", together with the "encounter dilution effect", is considered among the most important factors determining individual vigilance behavior. However, in addition to group size, other social and environmental factors also influence the degree of vigilance, including disturbance from human activities. In our study, we examined vigilance behavior of Khulans (Equus hemionus) in the Xinjiang Province in western China to test whether and how human disturbance and group size affect vigilance. According to our results, Khulan showed a negative correlation between group size and the percentage time spent vigilant, although this negative correlation depended on the groups' disturbance level. Khulan in the more disturbed area had a dampened benefit from increases in group size, compared to those in the undisturbed core areas. Provision of continuous areas of high-quality habitat for Khulans will allow them to form larger undisturbed aggregations and to gain foraging benefits through reduced individual vigilance, as well as anti-predator benefits through increased probability of predator detection.

  18. Field trials on the repellent activity of four plant products against mainly Mansonia population in western Ethiopia.

    PubMed

    Hadis, Mamuye; Lulu, Mesfin; Mekonnen, Yared; Asfaw, Teffera

    2003-03-01

    The repellent activity of essential oils of lemon eucalyptus (Eucalyptus maculata citrodion), rue (Ruta chalepensis), oleoresin of pyrethrum (Chrysanthemum cinerariaefolium) and neem (Azadiracta indica) have been field tested as 40%, 50% and 75% solutions in coconut oil against populations of mosquitoes consisting mainly of Mansonia in Gambella, western Ethiopia. A latin square design was used to randomize the test subjects for possible individual differences for mosquito attraction. Repellency was evaluated as the percentage protection. Deet was included in the study for comparison. All the plant products manifested repellency. At 50% concentration at which the highest repellency was recorded the protection was 91.6%, 87.0%, 96.0%, 97.9% for rue, neem, pyrethrum and deet, respectively. The essential oil of lemon eucalyptus was not tried at this concentration. At a 40% concentration deet, lemon eucalyptus and pyrethrum were significantly (p < 0.05) more effective than rue and neem. At a 50% concentration, deet and pyrethrum were significantly better (p < 0.05) than rue and neem. At a 75% concentration concentration, deet and lemon eucalypus performed significantly better (p < 0.05) than pyrethrum and neem. The difference between pyrethrum and neem was also significant (p < 0.01). PMID:12672146

  19. Biogeochemistry of a low-activity cold seep in the Larsen B area, western Weddell Sea, Antarctica

    NASA Astrophysics Data System (ADS)

    Niemann, H.; Fischer, D.; Graffe, D.; Knittel, K.; Montiel, A.; Heilmayer, O.; Nöthen, K.; Pape, T.; Kasten, S.; Bohrmann, G.; Boetius, A.; Gutt, J.

    2009-06-01

    First videographic indication of an Antarctic cold seep ecosystem was recently obtained from the collapsed Larsen B ice shelf, western Weddell Sea (Domack et al., 2005). Within the framework of the R/V Polarstern expedition ANTXXIII-8, we revisited this area for geochemical, microbiological and further videographical examinations. During two dives with ROV Cherokee (MARUM, Bremen), several bivalve shell agglomerations of the seep-associated, chemo syntheticclam Calyptogena sp. were found in the trough of the Crane and Evans glacier. The absence of living clam specimens indicates that the flux of sulphide and hence the seepage activity is diminished at present. This impression was further substantiated by our geochemical observations. Concentrations of thermogenic methane were moderately elevated with 2 μM in surface sediments of a clam patch, increasing up to 9 μM at a sediment depth of about 1 m in the bottom sections of the sediment cores. This correlated with a moderate decrease in sulphate from 28 mM at the surface down to 23.4 mM, an increase in sulphide to up to 1.43 mM and elevated rates of the anaerobic oxidation of methane (AOM) of up to 600 pmol cm-3 d-1 at about 1 m below the seafloor. Molecular analyses indicate that methanotrophic archaea related to ANME-3 are the most likely candidates mediating AOM in sediments of the Larsen B seep (Domack et al., 2005; EOS 86, 269-276).

  20. Biogeochemistry of a low-activity cold seep in the Larsen B area, western Weddell Sea, Antarctica

    NASA Astrophysics Data System (ADS)

    Niemann, H.; Fischer, D.; Graffe, D.; Knittel, K.; Montiel, A.; Heilmayer, O.; Nöthen, K.; Pape, T.; Kasten, S.; Bohrmann, G.; Boetius, A.; Gutt, J.

    2009-11-01

    First videographic indication of an Antarctic cold seep ecosystem was recently obtained from the collapsed Larsen B ice shelf, western Weddell Sea (Domack et al., 2005). Within the framework of the R/V Polarstern expedition ANTXXIII-8, we revisited this area for geochemical, microbiological and further videographical examinations. During two dives with ROV Cherokee (MARUM, Bremen), several bivalve shell agglomerations of the seep-associated, chemosynthetic clam Calyptogena sp. were found in the trough of the Crane and Evans glacier. The absence of living clam specimens indicates that the flux of sulphide and hence the seepage activity is diminished at present. This impression was further substantiated by our geochemical observations. Concentrations of thermogenic methane were moderately elevated with 2 μM in surface sediments of a clam patch, increasing up to 9 μM at a sediment depth of about 1 m in the bottom sections of the sediment cores. This correlated with a moderate decrease in sulphate from about 28 mM at the surface down to 23.4 mM, an increase in sulphide to up to 1.43 mM and elevated rates of the anaerobic oxidation of methane (AOM) of up to 600 pmol cm-3 d-1 at about 1 m below the seafloor. Molecular analyses indicate that methanotrophic archaea related to ANME-3 are the most likely candidates mediating AOM in sediments of the Larsen B seep.

  1. Sandfly species diversity in association with human activities in the Kani tribe settlements of the Western Ghats, Thiruvananthapuram, Kerala, India

    PubMed Central

    Ranganathan, Srinivasan; Swaminathan, Subramanian

    2015-01-01

    Sandfly prevalence in the Kani tribe settlements of Western Ghats in India was investigated. A total of 1,279 sandflies comprising 17 species was obtained. Sandfly abundance showed a negative correlation (r = -0.97, p = 0.003) with increase in altitudinal ranges from 0-1,000 m. When sandfly samples were grouped according to landscape characteristics of the location, the estimated Shannon-Weiner index (H) and species richness index (S) were high and species evenness index (J) was low in settlements located at 0-300 m altitudinal range. On the contrary, the values of H and J were high, while S was low at 301-600 m altitudinal range. With further increase in altitude, species diversity, S and J were low. Though the relative abundance of sandflies decreased with increase in altitude, the influence of altitudinal variation could not be attributed to determine sandfly diversity, since the number of sampling units were not uniform at all the altitudinal gradients due to nonavailability of suitable resting shelters. Sandfly species showed great aggregation at 0-300 m altitude interval, where not only the number of settlements were maximum (n = 19), but also the environmental conditions favoured sandfly abundance due to the concentration of tribal settlements, human dwellings and his activities. PMID:25946240

  2. Human Activity Dampens the Benefits of Group Size on Vigilance in Khulan (Equus hemionus) in Western China

    PubMed Central

    Wang, Mu-Yang; Ruckstuhl, Kathreen E.; Xu, Wen-Xuan; Blank, David; Yang, Wei-Kang

    2016-01-01

    Animals receive anti-predator benefits from social behavior. As part of a group, individuals spend less time being vigilant, and vigilance decreases with increasing group size. This phenomenon, called “the many-eyes effect”, together with the “encounter dilution effect”, is considered among the most important factors determining individual vigilance behavior. However, in addition to group size, other social and environmental factors also influence the degree of vigilance, including disturbance from human activities. In our study, we examined vigilance behavior of Khulans (Equus hemionus) in the Xinjiang Province in western China to test whether and how human disturbance and group size affect vigilance. According to our results, Khulan showed a negative correlation between group size and the percentage time spent vigilant, although this negative correlation depended on the groups’ disturbance level. Khulan in the more disturbed area had a dampened benefit from increases in group size, compared to those in the undisturbed core areas. Provision of continuous areas of high-quality habitat for Khulans will allow them to form larger undisturbed aggregations and to gain foraging benefits through reduced individual vigilance, as well as anti-predator benefits through increased probability of predator detection. PMID:26756993

  3. Sandfly species diversity in association with human activities in the Kani tribe settlements of the Western Ghats, Thiruvananthapuram, Kerala, India.

    PubMed

    Ranganathan, Srinivasan; Swaminathan, Subramanian

    2015-04-01

    Sandfly prevalence in the Kani tribe settlements of Western Ghats in India was investigated. A total of 1,279 sandflies comprising 17 species was obtained. Sandfly abundance showed a negative correlation (r = -0.97, p = 0.003) with increase in altitudinal ranges from 0-1,000 m. When sandfly samples were grouped according to landscape characteristics of the location, the estimated Shannon-Weiner index (H) and species richness index (S) were high and species evenness index (J) was low in settlements located at 0-300 m altitudinal range. On the contrary, the values of H and J were high, while S was low at 301-600 m altitudinal range. With further increase in altitude, species diversity, S and J were low. Though the relative abundance of sandflies decreased with increase in altitude, the influence of altitudinal variation could not be attributed to determine sandfly diversity, since the number of sampling units were not uniform at all the altitudinal gradients due to nonavailability of suitable resting shelters. Sandfly species showed great aggregation at 0-300 m altitude interval, where not only the number of settlements were maximum (n = 19), but also the environmental conditions favoured sandfly abundance due to the concentration of tribal settlements, human dwellings and his activities. PMID:25946240

  4. BLOT: A mesh and curve plot program for the output of a finite element analysis

    SciTech Connect

    Gilkey, A.P.; Glick, J.H.

    1989-06-01

    BLOT is a graphics program for post-processing of finite element analysis output that is presented in the EXODUS database format. It is command driven with free-format input and can drive any graphics device supported by the Sandia Virtual Device Interface. BLOT produces mesh plots with various representations of the analysis output variables. The major mesh plot capabilities are deformed mesh plots, line contours, banded contours, vector plots of two or three variables (e.g., velocity vectors), and symbol plots of scalar variables (e.g., temperature). Pathlines of analysis variables can also be drawn on the mesh. BLOT's features include element selection by material, element birth and death, multiple views for combining several displays on each plot, symmetry mirroring, and node and element numbering. BLOT can also produce X-Y curve plots of the analysis variables. BLOT generates time-versus-variable plots or variable-versus-variable plots. It also generates distance versus-variable plots at selected time steps where the distance is the accumulated distance between pairs of nodes or element centers. 14 refs.

  5. Composition and antimicrobial activity of the essential oil of Heracleum thomsonii (Clarke) from the cold desert of the western Himalayas.

    PubMed

    Guleria, Shailja; Saini, Rikki; Jaitak, Vikas; Kaul, V K; Lal, Brij; Rahi, Praveen; Gulati, Arvind; Singh, Bikram

    2011-08-01

    Volatile oil composition of hydro-distilled (HD) and supercritical carbon dioxide (SC-CO(2)) essential oil of freshly collected aerial parts of Heracleum thomsonii (Umbeliferae) from the western Himalayas was studied by GC-FID and GC-MS. Results revealed qualitative and quantitative dissimilarity in the composition of hydro-distilled and SC-CO(2) extracted oils. Nineteen constituents, which accounted for 89.32% of total constituents in HD oil, represented by limonene (4.31%), (Z)-β-ocimene (3.69%), terpinolene (22.24%), neryl acetate (36.19%), nerol (9.51%) and p-cymene-8-ol (2.61%) were identified. In SC-CO(2) extracted oil, 24 constituents representing 89.95% of total constituents were identified. Terpinolene (5.08%), germacrene D (2.17%), neryl acetate (51.62%), nerol (9.78%), geranyl acetate (2.06%), α-bisabolol (2.48%) and 1-nonadecanol (4.96%) were the dominating constituents. In vitro antimicrobial activity of hydro-distilled oil was conducted against microrobial strains including two Gram-positive (Staphylococcus aureus and Bacillus subtilis) and five Gram-negative (Burkholderia cepacia, Escherichia coli, Enterobacter cloacae, Klebseilla pneumoniae and Pseudomonas aeruginosa) bacteria as well as seven fungi (Candida albicans, Issatchenkia orientalis, Aspergillus flavus, Aspergillus niger, Aspergillus parasiticus, Aspergillus sydowii and Trichophyton rubrum) using broth microdilution method. The results of bioassay showed that the oil exhibited moderate to high antimicrobial activity against fungi C. albicans (MIC 625 µg ml(-1)), A. parasiticus (MIC 312.5 µg ml(-1)), A. sydowii (MIC 312.5 µg ml(-1)), T. rubrum (MIC 625 µg ml(-1)), Gram-positive bacteria B. subtilis (MIC 625 µg ml(-1)) and Gram-negative bacteria P. aeruginosa (MIC 312.5 µg ml(-1)). PMID:21854172

  6. Dissolved phosphorus pools and alkaline phosphatase activity in the euphotic zone of the Western north pacific ocean.

    PubMed

    Suzumura, Masahiro; Hashihama, Fuminori; Yamada, Namiha; Kinouchi, Shinko

    2012-01-01

    We measured pools of dissolved phosphorus (P), including dissolved inorganic P (DIP), dissolved organic P (DOP) and alkaline phosphatase (AP)-hydrolyzable labile DOP (L-DOP), and kinetic parameters of AP activity (APA) in the euphotic zone in the western North Pacific Ocean. Samples were collected from one coastal station in Sagami Bay, Japan, and three offshore stations between the North Pacific subtropical gyre (NPSG) and the Kuroshio region. Although DIP concentrations in the euphotic zone at all stations were equally low, around the nominal method detection limit of 20 nmol L(-1), chlorophyll a (Chl a) concentrations were one order of magnitude greater at the coastal station. DOP was the dominant P pool, comprising 62-92% of total dissolved P at and above the Chl a maximum layer (CML). L-DOP represented 22-39% of the total DOP at the offshore stations, whereas it accounted for a much higher proportion (about 85%) in the coastal surface layers. Significant correlations between maximum potential AP hydrolysis rates and DIP concentrations or bacterial cell abundance in the offshore euphotic zone suggest that major APA in the oligotrophic surface ocean is from bacterial activity and regulated largely by DIP availability. Although the range of maximum potential APA was comparable among the environmental conditions, the in situ hydrolysis rate of L-DOP in the coastal station was 10 times those in the offshore stations. L-DOP turnover time at the CML ranged from 4.5 days at the coastal station to 84.4 days in the NPSG. The ratio of the APA half-saturation constant to the ambient L-DOP concentration decreased markedly from the NPSG to the coastal station. There were substantial differences in the rate and efficiency of DOP remineralization and its contribution as the potential P source between the low-phosphate/high-biomass coastal ecosystem and the low-phosphate/low biomass oligotrophic ocean. PMID:22457661

  7. Dissolved phosphorus pools and alkaline phosphatase activity in the euphotic zone of the Western north pacific ocean.

    PubMed

    Suzumura, Masahiro; Hashihama, Fuminori; Yamada, Namiha; Kinouchi, Shinko

    2012-01-01

    We measured pools of dissolved phosphorus (P), including dissolved inorganic P (DIP), dissolved organic P (DOP) and alkaline phosphatase (AP)-hydrolyzable labile DOP (L-DOP), and kinetic parameters of AP activity (APA) in the euphotic zone in the western North Pacific Ocean. Samples were collected from one coastal station in Sagami Bay, Japan, and three offshore stations between the North Pacific subtropical gyre (NPSG) and the Kuroshio region. Although DIP concentrations in the euphotic zone at all stations were equally low, around the nominal method detection limit of 20 nmol L(-1), chlorophyll a (Chl a) concentrations were one order of magnitude greater at the coastal station. DOP was the dominant P pool, comprising 62-92% of total dissolved P at and above the Chl a maximum layer (CML). L-DOP represented 22-39% of the total DOP at the offshore stations, whereas it accounted for a much higher proportion (about 85%) in the coastal surface layers. Significant correlations between maximum potential AP hydrolysis rates and DIP concentrations or bacterial cell abundance in the offshore euphotic zone suggest that major APA in the oligotrophic surface ocean is from bacterial activity and regulated largely by DIP availability. Although the range of maximum potential APA was comparable among the environmental conditions, the in situ hydrolysis rate of L-DOP in the coastal station was 10 times those in the offshore stations. L-DOP turnover time at the CML ranged from 4.5 days at the coastal station to 84.4 days in the NPSG. The ratio of the APA half-saturation constant to the ambient L-DOP concentration decreased markedly from the NPSG to the coastal station. There were substantial differences in the rate and efficiency of DOP remineralization and its contribution as the potential P source between the low-phosphate/high-biomass coastal ecosystem and the low-phosphate/low biomass oligotrophic ocean.

  8. Active Extensional Faulting at the Southern Half-Graben Belt of the Tepic-Zacoalco Rift, Western Mexico

    NASA Astrophysics Data System (ADS)

    Rosas-Elguera, J.; Ferrari, L.; Delgado, M.; Uribe, A.; Valdivia, L.; Castillo, R.

    2003-12-01

    In the past decade much debate has centered upon the kinematics and the mechanism of continental deformation in western Mexico and the motion of the Jalisco block relative to North America. Two distinct models have been proposed. The first one suggest a NW-motion of the Jalisco block that would implies a right-lateral faulting along the Tepic-Zacoalco rift (TZR). More recently others authors have documented a N-NE extensional tectonics active since late Miocene and suggested that the continental boundaries of the Jalisco block, are older structures reactivated by plate boundary forces. Studies on the crustal seismicity and the kinematics of Quaternary faults provide another constraint on the direction of motion between the Jalisco block and North America. On November 4, 5, 6, and 7, 1995, one month after the October 09, 1995, Manzanillo earthquake (Mw = 8.0), a swarm of small events was felt in the Amatlan de Ca¤as half-graben and recorded by the regional seismic network of Comision Federal de Electricidad. The coda magnitude of the largest event was Mc = 2.5-3.6 and the events were located depth ranging from 6 to 10 km. This seismic activity provoked that people from Pie de la Cuesta and Yerbabuena villages were evacuated. After that a seismic station equipped with an analogic seismograph MEQ-800 at Pie de la Cuesta was installed for three months. During the same time, October, 1995, some houses distributed along a WNW trend in Ameca city underwent severe damages, they are. The digital elevations model of the Ameca city suggest that several structures tectonics are shorter than 2 km are present in the area. The present direction of motion of the Rivera plate relative to North America plate along Middle America Trench has been estimated between N19° E to N48° E (e.g. Bandy et al., 1996). During the October 09, 1995, subduction-related earthquake (Mw = 8.0) a GPS network recorded a SW motion of the Jalisco block which could be associated to an elastic deformation

  9. Characterization of the spectrum of insecticidal activity of a double-stranded RNA with targeted activity against Western Corn Rootworm (Diabrotica virgifera virgifera LeConte).

    PubMed

    Bachman, Pamela M; Bolognesi, Renata; Moar, William J; Mueller, Geoffrey M; Paradise, Mark S; Ramaseshadri, Parthasarathy; Tan, Jianguo; Uffman, Joshua P; Warren, Joanne; Wiggins, B Elizabeth; Levine, Steven L

    2013-12-01

    The sequence specificity of the endogenous RNA interference pathway allows targeted suppression of genes essential for insect survival and enables the development of durable and efficacious insecticidal products having a low likelihood to adversely impact non-target organisms. The spectrum of insecticidal activity of a 240 nucleotide (nt) dsRNA targeting the Snf7 ortholog in Western Corn Rootworm (WCR; Diabrotica virgifera virgifera) was characterized by selecting and testing insects based upon their phylogenetic relatedness to WCR. Insect species, representing 10 families and 4 Orders, were evaluated in subchronic or chronic diet bioassays that measured potential lethal and sublethal effects. When a specific species could not be tested in diet bioassays, the ortholog to the WCR Snf7 gene (DvSnf7) was cloned and corresponding dsRNAs were tested against WCR and Colorado potato beetle (Leptinotarsa decemlineata); model systems known to be sensitive to ingested dsRNA. Bioassay results demonstrate that the spectrum of activity for DvSnf7 is narrow and activity is only evident in a subset of beetles within the Galerucinae subfamily of Chrysomelidae (>90% identity with WCR Snf7 240 nt). This approach allowed for evaluating the relationship between minimum shared nt sequence length and activity. A shared sequence length of ≥ 21 nt was required for efficacy against WCR (containing 221 potential 21-nt matches) and all active orthologs contained at least three 21 nt matches. These results also suggest that WCR resistance to DvSnf7 dsRNA due to single nucleotide polymorphisms in the target sequence of 240 nt is highly unlikely. PMID:23748931

  10. Characterization of the spectrum of insecticidal activity of a double-stranded RNA with targeted activity against Western Corn Rootworm (Diabrotica virgifera virgifera LeConte).

    PubMed

    Bachman, Pamela M; Bolognesi, Renata; Moar, William J; Mueller, Geoffrey M; Paradise, Mark S; Ramaseshadri, Parthasarathy; Tan, Jianguo; Uffman, Joshua P; Warren, Joanne; Wiggins, B Elizabeth; Levine, Steven L

    2013-12-01

    The sequence specificity of the endogenous RNA interference pathway allows targeted suppression of genes essential for insect survival and enables the development of durable and efficacious insecticidal products having a low likelihood to adversely impact non-target organisms. The spectrum of insecticidal activity of a 240 nucleotide (nt) dsRNA targeting the Snf7 ortholog in Western Corn Rootworm (WCR; Diabrotica virgifera virgifera) was characterized by selecting and testing insects based upon their phylogenetic relatedness to WCR. Insect species, representing 10 families and 4 Orders, were evaluated in subchronic or chronic diet bioassays that measured potential lethal and sublethal effects. When a specific species could not be tested in diet bioassays, the ortholog to the WCR Snf7 gene (DvSnf7) was cloned and corresponding dsRNAs were tested against WCR and Colorado potato beetle (Leptinotarsa decemlineata); model systems known to be sensitive to ingested dsRNA. Bioassay results demonstrate that the spectrum of activity for DvSnf7 is narrow and activity is only evident in a subset of beetles within the Galerucinae subfamily of Chrysomelidae (>90% identity with WCR Snf7 240 nt). This approach allowed for evaluating the relationship between minimum shared nt sequence length and activity. A shared sequence length of ≥ 21 nt was required for efficacy against WCR (containing 221 potential 21-nt matches) and all active orthologs contained at least three 21 nt matches. These results also suggest that WCR resistance to DvSnf7 dsRNA due to single nucleotide polymorphisms in the target sequence of 240 nt is highly unlikely.

  11. Field evidence for an active transtensional basin in western Anatolia: The Inner Bay of İzmir

    NASA Astrophysics Data System (ADS)

    Uzel, B.; Sözbilir, H.; Sümer, Ö.; Inci, U.; Özkaymak, Ç.

    2009-04-01

    The west Anatolian extensional province is dominantly shaped by an E-W-trending graben-forming high-angle normal faults. However, there are some studies revealing the presence of a number of NE-trending strike-slip faults deforming the western Anatolia crust onshore and offsore. The Izmir Bay and the surrounding area is located at the NE-trending strike-slip dominated zone of weakness known as the Izmir-Balıkesir Transfer Zone forming the western end of the west Anatolian extensional province. The Izmir Bay is a marine basin that controlled by NE, NW, N-S and E-W trending active faults. The bay of Izmir is topographically divided into an E-trending inner bay and an NW-trending outer bay. The southern sector of the inner bay is bounded by ˙Izmir Fault which is made up of two main segments trending approximately in E-W. According to field studies, the ˙Izmir Fault is an oblique-slip normal fault dipping to the north. The northern sector of the inner bay is bounded by Kaŗsıyaka Fault which is antithetic to the Izmir Fault. Kaŗsıyaka Fault in this section has a characteristic of oblique-slip normal fault dipping to the south. The NE-trending Seferihisar Fault, which is approximately developed normal to the ˙Izmir Fault is approximately 30 km long on the land. Morphological, geological and seismological evidences suggest that the northern segment of the Seferihisar fault has been activated during Holocene time. The fault is a dextral strike-slip fault with oblique-slip movement. The fault trends in N20E at the southern part where it cuts the Upper Cretaceous Bornova Flysch and Quaternary alluvium. Towards the north, the strike of the fault changes to N40-50E and it gets connected with the ˙Izmir Fault. Within the excavated trenches on the fault segments mapped in 1/1000 scale, it is established that the strike-slip fault (Seferihisar Fault) is transformed by bending into the normal fault (Izmir Fault). Kinematic analysis carried out at 9 stations suggest that

  12. The Modulation of Tropical Storm Activity in the Western North Pacific by the Madden-Julian Oscillation in GEOS-5 AGCM Experiments

    NASA Technical Reports Server (NTRS)

    Kim, Dongmin; Lee, Myong-In; Kim, Hye-Mi; Schubert, Siegfried D.; Yoo, Jin H.

    2014-01-01

    This study examines the influence of the Madden-Julian Oscillation (MJO) on tropical storm (TS) activity in the western North Pacific, using observations and GEOS-5 simulations at 50-km horizontal resolution. While GEOS-5 produces an MJO of faster propagation and weaker amplitude, it nevertheless reproduces the observed modulation of TS activity by the MJO with the highest TS genesis and increased track density in the active phases of MJO. The study suggests that the simulation of the sub-seasonal variability of TS activity could be improved by improving the simulations of the MJO in climate models.

  13. 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.

  14. Evaluation of a Treponema pallidum western immunoblot assay as a confirmatory test for syphilis.

    PubMed Central

    Byrne, R E; Laska, S; Bell, M; Larson, D; Phillips, J; Todd, J

    1992-01-01

    Tests for the detection of antibodies to Treponema pallidum are recommended for the confirmation of reactive nontreponemal test results and the accurate diagnosis of syphilis. The present-day use of Western blot (immunoblot) technology for the diagnosis of retroviruses prompted the development and evaluation of a Western blot assay with whole-cell T. pallidum as the antigen. The assay detected antibodies in syphilitic serum or plasma from dilutions of specimens incubated overnight with test strips. A test was considered positive when at least three of four major antigens having molecular masses of 15.5, 17, 44.5, and 47 kDa were detected. The Western blot assay had 93.8% sensitivity and 100% specificity for clinically defined samples. The Western blot assay was compared with double-staining fluorescent treponemal antibody absorption [FTA-ABS (DS)], which had a sensitivity and a specificity of 91.7 and 92.0%, respectively. Dilution series studies of syphilis-positive specimens indicated that the Western blot assay has an endpoint of reactivity at least 3 to 4 serial dilutions greater than that for FTA-ABS (DS). Overall, the greater than 95% agreement between the Western blot assay and FTA-ABS (DS) for clinically defined specimens indicates that the sensitivity of the Western blot assay is equal to or greater than that of FTA-ABS (DS). The Western blot assay demonstrated no false-positive or equivocal reactivities for nonsyphilitic specimens, including normal specimens (both plasma and serum), biological false-positives, and specimens with elevated gamma globulin or antinuclear antibody. We conclude that the high sensitivity and specificity of the T. pallidum Western blot assay, together with its simplicity and objectivity, make it a good confirmatory test for syphilis. Images PMID:1734042

  15. Mass transport deposits as witness of Holocene seismic activity on the Ligurian margin, Western Mediterranean (ASTARTE project)

    NASA Astrophysics Data System (ADS)

    Samalens, Kevin; Cattaneo, Antonio; Migeon, Sébastien

    2016-04-01

    The Ligurian Margin (Western Mediterranean) is at the transition between the Southern Alpes and the Liguro-Provençal margin and it is one of the most seismic areas of France. Several historic earthquakes have been indexed; the strongest, on February 23rd, 1887, occurred offshore Menton and Imperia and also caused a tsunami wave. Its equivalent magnitude has been estimated between 6 and 6.5. In addition, a moderate recurrent seismicity shakes the margin. The aim of this study is to understand the link between seismic activity and slope destabilization, and to identify the sedimentary deposits resulting from mass transport or turbidity currents. During Malisar (Geoazur laboratory), Prisme 2 and Prisme 3 (Ifremer) cruises, bathymetry, seafloor imagery (SAR), geophysics data (CHIRP SYSIF and high resolution seismics), and sediment cores have been acquired on the continental slope, focussing on canyons and submarine landslides, and in the basin. These data record numerous mass transport deposits (slump, debrites) in the different physiographic areas of the margin. To search for evidences of past Ligurian margin seismicity during the Holocene, we focused on the northeast part of the margin, the Finale area. We identified and sampled acoustically transparent Mass Transport Deposits up to 20-m thick in the bottom of three coaleshing canyons: Noli, Pora and Centa canyons from W to E in the area offshore Finale Ligure. We also recovered an MTD in the collecting deeper canyon system. MTDs in cores appear as sediment with different degrees of deformation (tilted blocks, slump, debrites) and are topped by hemipelagites. The radiocarbon age of the top of MTDs can be considered synchronous and centered around 4900 yr BP. Mass wasting occurring over more than 50 km of the Ligurian margin could indicate that an earthquake stroke the Finale area sector at that time.

  16. Cycling of Dissolved Organic Phosphorus and Alkaline Phosphatase Activity in Euphotic Zone of the Western North Pacific

    NASA Astrophysics Data System (ADS)

    Suzumura, M.

    2010-12-01

    Phosphorus is an essential nutrient for marine organisms. In oligotrophic environments, concentrations of dissolved inorganic phosphate (SRP), the most bioavailable form of phosphorus, are low and have been hypothesized to constrain the primary productivity. Evidence has been found that dissolved organic phosphorus (DOP) supports a significant fraction of primary production through hydrolytic remineralization of DOP to SRP by alkaline phosphatase (APA). In this study, DOP biogeochemistry was investigated at three locations of the open-ocean environment in the Kuroshio region and at a semi-eutrophic coastal site of the western North Pacific. Concentrations of SRP, DOP and hydrolyzable ester-P were measured in the euphotic zone. Kinetic parameters of APA were determined using a fluorogenic substrate, including potential maximum velocity (Vmax), apparent Michaelis-Menten half-saturation constant (Km), and turnover time (TA) of APA hydrolyzable DOP. SRP concentrations were quite low (≤ 10 nM) in the surface seawater and rapidly increased below the chlorophyll a maximum layer (CML). DOP concentration ranged from 29 to 223 nM. Above the CML, DOP composed a major fraction accounting for 60-100% of dissolved total P. A significant linear relationship was found between the concentrations of SRP and hydrolyzable ester-P (R2 = 0.83, P < 0.01). This suggests active utilization of ester-P under phosphate-depleted conditions. In the Kuroshio region, Vmax of APA exhibited the highest value at the surface water (0 m) and decreased rapidly with depth, while at the coastal site the peak value was found at CML. TA of hydrolyzable DOP was quite variable among the locations and increased with depth especially below CML. The estimated values of in situ hydrolysis rate were much lower (2-34%) than the potential Vmax which was determined with the addition of an excess amount of the substrate. The results suggest that marine microbes can efficiently and rapidly utilize hydrolyzable DOP

  17. Identification of additional MAP kinases activated upon PAMP treatment

    PubMed Central

    Nitta, Yukino; Ding, Pingtao; Zhang, Yuelin

    2014-01-01

    Mitogen-activated protein (MAP) kinase cascades play important roles in plant immunity. Upon pathogen associated molecular pattern (PAMP) treatment, MPK3, MPK6 and MPK4 are quickly activated by upstream MKKs through phosphorylation. Western blot analysis using α-phospho-p44/42-ERK antibody suggests that additional MPKs with similar size as MPK4 are also activated upon PAMP perception. To identify these MAP kinases, 7 candidate MPKs with similar sizes as MPK4 were selected for further analysis. Transgenic plants expressing these MPKs with a ZZ-3xFLAG double tag of 17 kD were generated and analyzed by western blot. MPK1, MPK11 and MPK13 were found to be phosphorylated upon treatment with flg22. Our study revealed additional MAPKs being activated during PAMP-triggered immunity. PMID:25482788

  18. The two-fold sea surface temperature change associated with interannual cyclone activities in the Western North Pacific

    NASA Astrophysics Data System (ADS)

    Chang, C.

    2013-05-01

    Concurrent with most large El Nino events, cold Sea Surface Temperature anomalies are observed over the western Pacific region. Composite of 14 El Nino events during 1955-2003 reveals that there are two stage development of sea surface temperature cooling during the El Nino year: the first peak of cooling appears in August-September of Nino(0) year, while the second peak in January-February of Nino(1) year. A few findings suggested that the local air-sea interaction feedback is potent in this region (Wang et al., 2001; Hsu et al., 2001). It is proposed that the increase in evaporation cooling associated with the presence of the anomalous atmospheric anticyclone during the El Nino winter leads to cold sea surface temperature anomalies in the Philippine Sea, in turn the oceanic anomalies feedback on the overlying atmospheric circulation. The wind-evaporation-sea surface temperature (WES) feedback (Xie and Philander, 1994) seems to be the major mechanism for the sea surface temperature cooling during the second peak when the atmospheric anticyclone anomaly straddling the Philippine Sea and South China Sea appears, causing warming(cooling) in the South China Sea(Philippine Sea) (Wang et al., 2003). However, the formation of the cooling in August and September (the first cooling peak) is not clear. The impact of cyclone activities on the regional sea surface temperature change during the first peak is investigated. The time scale of the sea surface temperature restoring time following tropical cyclones is approximately 30-35 days for a tropical storm to 50-60 days for a category 3-5 hurricane, with significant storm-specific and basin-specific variability (Hart et al., 2007). It is also known that there is a tendency in El Niño years toward tropical cyclones that are both more intense and longer-lived than in La Niña years (Carmargo et al., 2004). The long-lasting of cyclones' self-induced ocean cooling would reflect onto the monthly average; in a sense that for the

  19. The Seismic Broad Band Western Mediterranean (wm) Network and the Obs Fomar Pool: Current state and Obs activities.

    NASA Astrophysics Data System (ADS)

    Pazos, Antonio; Davila, Jose Martin; Buforn, Elisa; Bezzeghoud, Mourad; Harnafi, Mimoun; Mattesini, Mauricio; Caldeira, Bento; Hanka, Winfried; El Moudnib, Lahcen; Strollo, Angelo; Roca, Antoni; Lopez de Mesa, Mireya; Dahm, Torsten; Cabieces, Roberto

    2016-04-01

    The Western Mediterranean (WM) seismic network started in 1996 as an initiative of the Royal Spanish Navy Observatory (ROA) and the Universidad Complutense de Madrid (UCM), with the collaboration of the GeoForschungsZentrum (GFZ) of Potsdam. A first broad band seismic station (SFUC) was installed close to Cádiz (South Spain). Since then, additional stations have been installed in the Ibero-Moghrebian region. In 2005, the "WM" code was assigned by the FDSN and new partners were jointed: Evora University (UEVO, Portugal), the Scientifique Institute of Rabat (ISRABAT, Morocco), and GFZ. Now days, the WM network is composed by 15 BB stations, all of them with Streckaisen STS-2 or STS-2.5 sensors, Quanterra or Earthdata digitizers and SeiscomP. Most them have co-installed a permanent geodetic GPS stations, and some them also have an accelerometer. There are 10 stations deployed in Spanish territory (5 in the Iberian peninsula, 1 in Balearic islands and 4 in North Africa Spanish places) with VSAT or Internet communications, 2 in Portugal (one of them without real time), and 3 in Morocco (2 VSAT and 1 ADSL). Additionally, 2 more stations (one in South Spain and one in Morocco) will be installed along this year. Additionally ROA has deployed a permanent real time VBB (CMG-3T: 360s) station at the Alboran Island. Due to the fact that part of the seismic activity is located at marine areas, and also because of the poor geographic azimuthal coverage at some zones provided by the land stations (specially in the SW of the San Vicente Cape area), ROA and UCM have acquired six broad band "LOBSTERN" OBS, manufactured by KUM (Kiel, Germany), conforming the OBS FOMAR pool. Three of them with CMG-40T sensor and the other with Trillium 120. These OBS were deployed along the Gibraltar strait since January to November 2014 to study the microseismicity in the Gibraltar strait area. In September 2015 FOMAR network has been deployed in SW of the San Vicente Cape for 8 months as a part of

  20. Airborne Laser Scanning (ALS) point cloud ground filtering for area of an active landslide (Doren, Western Austria)

    NASA Astrophysics Data System (ADS)

    Brodić, Nenad; Cvijetinović, Željko; Milenković, Milutin; Dorninger, Peter; Mitrović, Momir

    2014-05-01

    Ground filtering of point cloud is the primary step required for Digital Terrain Model (DTM) generation. The procedure is especially interesting for forested areas, since LiDAR systems can measure terrain elevation under vegetation cover with a high level of penetration. This work analyzes the potential of ALS data ground filtering for area of an active landslide. The results of ALS filtering, for example, may improve geomorphological and motion-detection studies. ALS data was collected during flight campaign 2011 under leaf-off conditions for Doren region, Vorarlberg, Western Austria. In this area, non-ground objects are mostly low vegetation such as shrubs, small trees etc. The vegetation is more dense in lower part of the landslide where erosion is smaller. Vegetation points can be removed based on the hypothesis that these are significantly higher than their neighboring points. However, in case of steep terrain, ground points may have the same heights as vegetation points, and thus, local slope should be considered. Also, if terrain roughness increases, the classification may become even more complex. Software system OPALS (Orientation and Processing of Airborne Laser Scanning data, Vienna University of Technology) was used for processing the ALS data. Labeling ground points has been made using physical and geometrical attributes (parameters) of ALS points. Also additional attributes were calculated in order to improve extraction. Since bare ground surface is usually smooth and continuous unlike vegetation, standard deviation of local elevations was used as roughness measure to differentiate these surfaces. EchoRatio (ER) was adopted as a measure of surface penetrability, while number of echoes and differentiation between echoes (EchoNumber) were also deployed in filtering. Since the ground points are measurements from bare-earth that are usually the lowest surface features in a local area, normalized height was defined as a rank of neighboring points

  1. Possible influence of western North Pacific monsoon on TC activity in mid-latitudes of East Asia

    NASA Astrophysics Data System (ADS)

    Choi, Ki-Seon; Cha, Yumi; Kim, Hae-Dong; Kang, Sung-Dae

    2016-01-01

    This study analyzed the correlation between tropical cyclone (TC) frequency and the Western North Pacific monsoon index (WNPMI), which have both been influential in East Asia's mid-latitude regions during the summer season over the past 37 years (1977-2013). A high positive correlation existed between these two variables, which was not reduced even if El Niño-Southern Oscillation (ENSO) years were excluded. To determine the cause of this positive correlation, the highest (positive WNPMI phase) and lowest WNPMIs (negative WNPMI phase) during a nine-year period were selected to analyze the mean difference between them, excluding ENSO years. In the positive WNPMI phase, TCs were mainly generated in the eastern seas of the tropical and subtropical western North Pacific, passing through the East China Sea and moving northward toward Korea and Japan. In the negative phase, TCs were mainly generated in the western seas of the tropical and subtropical western North Pacific, passing through the South China Sea and moving westward toward China's southern regions. Therefore, TC intensity in the positive phase was stronger due to the acquisition of sufficient energy from the sea while moving a long distance up to East Asia's mid-latitude. Additionally, TCs occurred more in the positive phase. Regarding the difference of the two phases between the 850 and 500-hPa streamlines, anomalous cyclones were strengthened in the tropical and subtropical western North Pacific, whereas anomalous anticyclones were strengthened in East Asia's mid-latitude regions. Due to these two anomalous pressure systems, anomalous southeasterlies developed in East Asia's mid-latitude regions, which played a role in the anomalous steering flows that moved TCs into these regions. Furthermore, due to the anomalous cyclones that developed in the tropical and subtropical western North Pacific, more TCs could be generated in the positive phase. Both the lower and upper tropospheric layers had warm anomalies

  2. Evaluation of climatological tropical cyclone activity over the western North Pacific in the CORDEX-East Asia multi-RCM simulations

    NASA Astrophysics Data System (ADS)

    Jin, Chun-Sil; Cha, Dong-Hyun; Lee, Dong-Kyou; Suh, Myoung-Seok; Hong, Song-You; Kang, Hyun-Suk; Ho, Chang-Hoi

    2016-08-01

    The ability of five regional climate models (RCMs), within the Coordinated Regional Climate Downscaling Experiment (CORDEX) for East Asia, to simulate tropical cyclone (TC) activity over the western North Pacific is evaluated. All RCMs are performed at ~50 km resolution over the CORDEX-East Asia domain, and are driven by the ECMWF Interim Re-Analysis (ERA-Interim) for the period 1989-2008. ERA-Interim sea surface temperature is prescribed as the lower boundary. Performances of the individual RCMs and multi-RCM ensemble mean are investigated in detail for 20-year climatology, intensity, and interannual variability of TC activity compared to observational datasets. Although most of the individual RCMs show significant biases and underestimate TC intensity due to horizontal resolutions still too low to resolve the most intense observed TCs, they reasonably capture the observed climatological spatial distribution and interannual variability of TC activity. The multi-RCM ensemble mean based on the model performance generally outperforms most of the individual models with smaller biases and higher correlation on the spatial and temporal variation of TC activity. This ensemble mean reduces the uncertainty in the simulated TC activity by a single RCM. These analyses suggest that the multi-RCM ensemble within CORDEX-East Asia can be applied to provide more reliable and credible estimation of future TC activity over the western North Pacific due to climate change.

  3. The squash blot technique and the detection of Leishmania major in Phlebotomus papatasi in Tunisia.

    PubMed

    Esseghir, S; Ftaïti, A; Ready, P D; Khadraoui, B; Zaafouri, B; Dellagi, K; Ben Ismaïl, R

    1993-01-01

    This study describes the preliminary applications of the squash blot technique in Tunisia, to detect Leishmania major in naturally infected Phlebotomus papatasi. 309 P. papatasi among 364 female sandflies squashed on to nylon Gene Screen DNA transfer membranes, were identified using the 3.2 kb ribosomal P. papatasi specific DNA probe described by Ready et al. A second hybridization using the Taq1 DNA probe described by Smith et al. (1989) allowed the detection and identification of the parasite in 15 (4.9%) of these P. papatasi specimens. The dissection of P. papatasi females during the same period and from the same biotopes showed an infection rate of 7.9% (9 positives among 113 dissected). The t proportion comparison test indicated that there is no significant statistical difference between the dissection and the squash blot technique for the estimation of infection rates of P. papatasi.

  4. Mapping Point Mutations in the Drosophila Rosy Locus Using Denaturing Gradient Gel Blots

    PubMed Central

    Gray, M.; Charpentier, A.; Walsh, K.; Wu, P.; Bender, W.

    1991-01-01

    Mutations within the rosy locus of Drosophila were mapped using blots of genomic DNA fragments separated on denaturing gradient gels. DNA sequence differences between otherwise identical small rosy DNA fragments were detected among the mutants as mobility shifts on the blots. Mutations were mapped to within a few hundred base pairs of rosy sequence in 100 of 130 mutants tested--a 77% detection rate. The sequence changes in 43 rosy mutations are presented; all but six of these were single base changes. Thirty-four of 36 sequenced mutations induced by the alkylating agents N-ethyl-N-nitrosourea and ethyl methanesulfonate were transitions. All of the mutations mapped in the rosy transcription unit. Twenty-three of the 43 sequenced mutations change the predicted rosy gene polypeptide sequence; the remainder would interrupt protein translation (17), or disrupt mRNA processing (3). PMID:1901817

  5. A Dual Color Southern Blot to Visualize Two Genomes or Genic Regions Simultaneously

    PubMed Central

    Zavala, Anamaria G.; Kulkarni, Amit S.; Fortunato, Elizabeth A.

    2014-01-01

    This report describes the development of a novel dual color Southern protocol to visualize two distinct genomes or genic regions simultaneously on a single Southern blot. The blot is developed with IRDye-conjugated antibody (Ab) and streptavidin that recognize Digoxigenin- (Dig) or biotin-labeled probes, respectively and visualized on an infrared imager. This protocol was validated by visualizing viral and host genomes of human cytomegalovirus (HCMV)-infected human fibroblasts. This technique utilizes extremely sensitive fluorescent imaging, allowing the detection of nanogram quantities of DNA, as opposed to microgram quantities needed in Southerns using radioactively labeled probes, and eliminates the inherent loss in signal after stripping and reprobing a Southern blot. The probes are labeled with non-radioactive Dig and biotin and can be stored for extended periods of time. This protocol will aid in studies of any system with two genomes, such as cells infected with numerous types of microorganisms (virus/parasites/bacteria), or studies of mitochondrial and nuclear DNA within the same cells. PMID:24389128

  6. Recently active reverse faulting in the Atacama Basin area, northern Chile: Implications for the distribution of convergence across the western South America plate boundary

    NASA Astrophysics Data System (ADS)

    Shyu, J. H.; Gonzalez, G.; Simons, M.; Aron, F.; Veloso, A.

    2007-12-01

    The western South American margin is one of the most active continental plate boundaries in the world. The ongoing convergence between the Nazca plate, or formerly the Farallon plate, and the South American plate produced the wide deformation belt of the Andes. In order to obtain more information about the active deformations in the central Andean belt to better understand the current distribution of convergence across the orogen, we attempted to map major structures that appear to be active recently by their topographic expressions using SRTM DEM and Landsat satellite images, followed by field observations. Results of our mapping show that there are many reverse faults that may be recently active in the area surrounding the Atacama Basin, in the Preandean Depression in northern Chile. These include a series of active reverse faults and related folds at the southeastern corner of the Atacama Basin, a major fold system that may be produced by an underlying fault just east of the basin, and a series of folds that forms the Cordillera de la Sal in the northern and western part of the basin. At the southeastern corner of the Atacama Basin, several geomorphic features indicate that at least some of the structures there have been active quite recently, including small drainages that cut through the folds and form active alluvial fans. Similar features of active river incision across folds are also present in the northern part of the basin. The fold system east of the basin may be one of the most important structures in the area. Deformed lava flows and deflected drainages indicate that this structure has been active recently, and growth strata near the fold suggest that it has been active for several myr. If so, the structure may be a major reverse fault system that defines the eastern boundary of the Atacama Basin, and may thus be an important onland structure that is responsible for absorbing part of the plate convergence.

  7. Imaging active faulting in a region of distributed deformation from the joint clustering of focal mechanisms and hypocentres: Application to the Azores-western Mediterranean region

    NASA Astrophysics Data System (ADS)

    Custódio, Susana; Lima, Vânia; Vales, Dina; Cesca, Simone; Carrilho, Fernando

    2016-04-01

    The matching between linear trends of hypocentres and fault planes indicated by focal mechanisms (FMs) is frequently used to infer the location and geometry of active faults. This practice works well in regions of fast lithospheric deformation, where earthquake patterns are clear and major structures accommodate the bulk of deformation, but typically fails in regions of slow and distributed deformation. We present a new joint FM and hypocentre cluster algorithm that is able to detect systematically the consistency between hypocentre lineations and FMs, even in regions of distributed deformation. We apply the method to the Azores-western Mediterranean region, with particular emphasis on western Iberia. The analysis relies on a compilation of hypocentres and FMs taken from regional and global earthquake catalogues, academic theses and technical reports, complemented by new FMs for western Iberia. The joint clustering algorithm images both well-known and new seismo-tectonic features. The Azores triple junction is characterised by FMs with vertical pressure (P) axes, in good agreement with the divergent setting, and the Iberian domain is characterised by NW-SE oriented P axes, indicating a response of the lithosphere to the ongoing oblique convergence between Nubia and Eurasia. Several earthquakes remain unclustered in the western Mediterranean domain, which may indicate a response to local stresses. The major regions of consistent faulting that we identify are the mid-Atlantic ridge, the Terceira rift, the Trans-Alboran shear zone and the north coast of Algeria. In addition, other smaller earthquake clusters present a good match between epicentre lineations and FM fault planes. These clusters may signal single active faults or wide zones of distributed but consistent faulting. Mainland Portugal is dominated by strike-slip earthquakes with fault planes coincident with the predominant NNE-SSW and WNW-ESE oriented earthquake lineations. Clusters offshore SW Iberia are

  8. The Geological, Geomorphological Features and Kinematic Analysis of Active Faults Controlling Kemalpaşa Basin, Southwestern Part of Gediz Graben, Western Anatolia

    NASA Astrophysics Data System (ADS)

    Tepe, Çiǧdem; Sözbilir, Hasan

    2016-04-01

    The purpose of this study is to discuss the geological and geomorphological features of active faults controlling Kemalpaşa Basin. The study consists of basin-bounding faults expressions, kinematic and geomorphic analysis. Kemalpaşa Basin, which is approximately ENE trending and asymmetric graben is located in the southern part of Gediz Graben. Menderes Massif and Bornova Complex comprise the basement rocks of basin. Kızılca Formation, Sütçüler Formation and Alluvium uncomformably overlie the basement rocks. Kemalpaşa Basin which is one of the Quaternary basin in the Western Anatolia Extensional Province was developed at the structural border of the Spildaǧı Fault Zone in the north and the Kemalpaşa Fault in the south. Both the north and south margin-bounding faults of Kemalpaşa Basin are oblique-slip normal faults. According to the results of kinematic analysis, Kemalpaşa Basin has been formed under a NE-GW trending extensional tectonic regime. The variation in the relative degree of tectonic activity in Kemalpaşa Basin and its surroundings were interpreted a detailed geomorphic study of the fault-generated mountain fronts and drainage pattern of the both sides. To identify the impacts of active faults controlling the north and south margins of Kemalpaşa Basin on the geomorphological evolution, the geomorphic indices such as drainage basin geometries, triangular facets, axial river profiles have been determined and the degree of tectonic activity in the both sides of Kemalpaşa Basin has been numerically defined using morphometric indexes such as asymmetry factor (AF), hypsometric curve and integral (HI), valley floor width-to-height ratio (Vf) and mountain front sinuosity (Smf). In morphometric analysis, the both sides of the basin were investigated separating into two segments as the west and east. The values of HI (0,28-0,60), Vf (0,27-0,60) and Smf (1,3) calculated for the western part of the north margin compared with the values of HI (0

  9. 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

  10. 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.

  11. Blotting assisted by heating and solvent extraction for DESI-MS imaging.

    PubMed

    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. PMID:23605686

  12. Blotting assisted by heating and solvent extraction for DESI-MS imaging.

    PubMed

    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.

  13. On The Relations Between Solar-Magnetospheric Activity, El-Nino Phenomenon And Tropical Cyclones Evolution In The North Western Part Of Pacific Ocean

    NASA Astrophysics Data System (ADS)

    Lazarev, A.

    2009-04-01

    We present results of the mutual statistical analysis of time series of Tropical Cyclones (TC) activity in the North-Western part of Pacific Ocean and (i) Solar activity and caused by it disturbances in the Earth's magnetosphere; (ii) Powerful climate forming factor - the El-Nino phenomenon. TC activity was characterized by the maximal and average speed of a wind in TC for a year. Solar activity was characterized by Wolf numbers. Solar-Magnetospheric activity was characterized by geomagnetic indices Аа and Ар. Activity of the El-Nino phenomenon was characterized by SOI-index. It was found that TC genesis in the South China Sea is subjected to the influence of Solar-Magnetospheric events - negative correlation between TC activity and Aа/Ap geomagnetic indices reaching value -0.6  0.1 for the period 1954-1978 with a zero time lag. Contrary, in the Philippine Sea Solar-Magnetospheric activity is a minor factor, due to compensation of incoming Solar radiation by the heat transfer of Passat current. In this region correlation between TC activity and SOI-index reaches value -0.64  0.1 for the interval 1961-1987 with a 2 year lag, approximately corresponding to the relaxation time of Sea Surface Temperature after El-Nino phenomenon.

  14. FY-09 Summary Report to the Office of Petroleum Reserves on the Western Energy Corridor Initiative Activities and Accomplishments

    SciTech Connect

    Thomas R. Wood

    2010-01-01

    To meet its programmatic obligations under the Energy Policy Act of 2005, the Office of Naval Petroleum and Shale Oil Reserves (NPSOR) has initiated the Western Energy Corridor Initiative (WECI). The WECI will implement the Unconventional Strategic Fuels Task Force recommendations for accelerating and promoting the development of domestic unconventional fuels to help meet the nations’ energy needs. The mission of the WECI is to bolster America’s future fuel security by facilitating socially and environmentally responsible development of unconventional fuels resources in the Western Energy Corridor, using sound engineering principles and science-based methods to define and assess benefits, impacts, uncertainties, and mitigation options and to resolve impediments. The Task Force proposed a three-year program in its commercialization plan. The work described herein represents work performed by Idaho National Laboratory (INL) in support of the DOE’s WECI. This effort represents an interim phase of work, designed to initiate only select portions of the initiative, limited by available funding resources within NPOSR. Specifically, the work presented here addresses what was accomplished in FY-09 with the remaining carryover (~$420K) from NPOSR FY-08 funds. It was the intent of the NPOSR program to seek additional funding for full implementation of the full scope of work; however, the original tasks were reduced in scope, terminated, or eliminated (as noted below). An effort is ongoing to obtain funding to continue the tasks initiated under this project. This study will focus on the integrated development of multiple energy resources in a carbon-neutral and environmentally acceptable manner. Emphasis will be placed on analyses of the interrelationships of various energy-resource development plans and the infrastructure, employment, training, fiscal, and economic demands placed on the region as a result of various development scenarios. The interactions at build

  15. 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.

  16. DMISA (dissociated membrane immunosorbent assay), a new ELISA technique performed with blotted samples.

    PubMed

    Guérin-Marchand, C; Batard, T; Brodard, V; Desvaux, F X; Sénéchal, H; David, B; Peltre, G

    1994-01-01

    This study describes the use of electrophoretically purified antigens blotted onto nitrocellulose, as solid phase antigens for enzyme-linked immunosorbent assays. This procedure is called DMISA, for dissociated membrane immunosorbent assay. The method is illustrated using immunoblotted antigens of Dactylis glomerata grass pollen extract. The band of interest was located on a print of nitrocellulose by light staining (India ink), then the corresponding strip of nitrocellulose was cut out. Immediately after its solubilization in ethyleneglycol monomethyl ether, the antigen coated nitrocellulose was precipitated by the addition of buffer. In this way the bulk of the antigen remained bound to the membrane. The resulting suspension was carefully washed, and used as a solid phase antigen in an enzyme-linked immunosorbent assay. Two different electrophoretic methods were used to separate the Dactylis glomerata antigens. We compared the results obtained with classical immunoblot and with DMISA, for IgG4 and IgE quantification using sera from patients allergic to D. glomerata and purified blotted antigens present at the nanogram level.

  17. Selective analysis of lipids by thin-layer chromatography blot matrix-assisted laser desorption/ionization imaging mass spectrometry.

    PubMed

    Zaima, Nobuhiro; Goto-Inoue, Naoko; Adachi, Kohsuke; Setou, Mitsutoshi

    2011-01-01

    Thin-layer chromatography (TLC) is an essential method for food composition analyses such as lipid nutrition analysis. TLC can be used to obtain information about the lipid composition of foods; however, it cannot be used for analyses at the molecular level. Recently we developed a new method that combines matrix-assisted laser desorption/ionization imaging mass spectrometry (MALDI-IMS) with TLC-blotting (TLC-Blot-MALDI-IMS). The combination of MALDI-IMS and TLC blotting enabled detailed and sensitive analyses of lipids. In this study, we applied TLC-Blot-MALDI-IMS for analysis of major phospholipids extracted from bluefin tuna. We showed that TLC-Blot-MALDI-IMS analysis could visualize and identify major phospholipids such as phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylcholine, and sphingomyelin.

  18. A comparison of bat activity at low and high elevations in the Black Hills of western Washington

    USGS Publications Warehouse

    Erickson, J.; Adams, Michael J.

    2003-01-01

    We examined the differences in activity patterns and community structure of bats between low (<150 m) and high ( ! 575 m) elevation sites in two habitats of the Capitol State Forest, Washington. Total bat activity averaged four times higher at low elevation sites than at high elevation sites. Feeding activity was almost 20 times higher at low elevation sites. However, the non-myotis group had similar activity levels at high and low elevation, whereas myotis group activity decreased at higher elevations. Different levels of activity between elevations could be the result of differences in insect availability, climatic conditions, and morphology of the bat species.

  19. Simulated impacts of two types of ENSO events on tropical cyclone activity in the western North Pacific: large-scale atmospheric response

    NASA Astrophysics Data System (ADS)

    Li, Chunxiang; Wang, Chunzai

    2014-05-01

    The present paper uses an atmospheric general circulation model to explore large-scale atmospheric response to various El Niño-Southern Oscillation events associated with tropical cyclone (TC) activity in the western North Pacific. The simulated response is basically consistent with and confirms the observed results. For eastern Pacific warm (EPW) event, anomalously wet ascent occurs over the tropical central/eastern Pacific and dry descent is over the western Pacific. This Walker circulation is associated with anomalous low-level convergence, reduced vertical wind shear (VWS), and enhanced genesis potential index (GPI) in the southeast sub-region. These are consistent with the observed increase of the TC formation in the southeast sub-region but decrease in the northwest sub-region during July-September (JAS) and the increase in the southwest and northwest sub-regions during October-December (OND). In addition, the strong westerly anomalies of the TC steering flow prevail in the East Asian coast, suppressing the TC northwestward or westward tracks. For eastern Pacific cold (EPC) event, all of the simulated variables show almost a mirror image of EPW. For central Pacific warm event, the anomalous Walker circulation shifts westward because of the westward shift of the maximum SST anomaly forcing. The anomalous subsidence associated with the western branch of the Walker circulation during OND shifts northward to the South China Seas, resulting in a decrease of the TC genesis there. The TC steering flow patterns during JAS are favorable for TCs to make landfall over Japan and Korea. Compared with EPC, the descending motion in the central/eastern Pacific is much stronger for central Pacific cold (CPC) event, accompanied by more enhanced VWS and reduced GPI in the southeast sub-region. Therefore, CPC provides a more adverse environment to the TC formation there during JAS and OND, consistent with the observed decrease of TC formation there. Moreover, the easterly

  20. Rotavirus detection by dot blot hybridization assay using a non-radioactive synthetic oligodeoxynucleotide probe.

    PubMed Central

    Fernández, J.; Sandino, A.; Yudelevich, A.; Avendaño, L. F.; Venegas, A.; Hinrichsen, V.; Spencer, E.

    1992-01-01

    A synthetic oligodeoxynucleotide of 40 nucleotides corresponding to nucleotides 33-72 of the gene coding for the viral protein VP7 of rotavirus, was used as a nucleic acid probe to develop a non-radioactive hybridization method for rotavirus detection. The probe was labelled at the 3' end with biotin-7-dATP. The sensitivity and specificity of the dot blot hybridization assay for rotavirus detection was evaluated with 303 stool specimens. The results indicate that the hybridization assay has a higher sensitivity than both PAGE and EIA. Among the rotavirus strains tested 37 different electropherotypes were found. The results suggest that rotavirus diagnosis by dot hybridization using a non-radioactive probe may become routine laboratory procedure because it is simple, highly specific and very sensitive. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 PMID:1312480

  1. PCR-Reverse Blot Hybridization Assay for Screening and Identification of Pathogens in Sepsis

    PubMed Central

    Choi, Yeonim; Wang, Hye-Young; Lee, Gyusang; Park, Soon-Deok; Jeon, Bo-Young; Uh, Young; Kim, Jong Bae

    2013-01-01

    Rapid and accurate identification of the pathogens involved in bloodstream infections is crucial for the prompt initiation of appropriate therapy, as this can decrease morbidity and mortality rates. A PCR-reverse blot hybridization assay for sepsis, the reverse blot hybridization assay (REBA) Sepsis-ID test, was developed; it uses pan-probes to distinguish Gram-positive and -negative bacteria and fungi. In addition, the assay was designed to identify bacteria and fungi using six genus-specific and 13 species-specific probes; it uses additional probes for antibiotic resistance genes, i.e., the mecA gene of methicillin-resistant Staphylococcus aureus (MRSA) and the vanA and vanB genes of vancomycin-resistant enterococci (VRE). The REBA Sepsis-ID test successfully identified clinical isolates and blood culture samples as containing Gram-positive bacteria, Gram-negative bacteria, or fungi. The results matched those obtained with conventional microbiological methods. For the REBA Sepsis-ID test, of the 115 blood culture samples tested, 47 (40.8%) and 49 (42.6%) samples were identified to the species and genus levels, respectively, and the remaining 19 samples (16.5%), which included five Gram-positive rods, were identified as Gram-positive bacteria, Gram-negative bacteria, or fungi. The antibiotic resistances of the MRSA and VRE strains were identified using both conventional microbiological methods and the REBA Sepsis-ID test. In conclusion, the REBA Sepsis-ID test developed for this study is a fast and reliable test for the identification of Gram-positive bacteria, Gram-negative bacteria, fungi, and antibiotic resistance genes (including mecA for MRSA and the vanA and vanB genes for VRE) in bloodstream infections. PMID:23447637

  2. Subduction of the South Chile active spreading ridge: A 17 Ma to 3 Ma magmatic record in central Patagonia (western edge of Meseta del Lago Buenos Aires, Argentina)

    NASA Astrophysics Data System (ADS)

    Boutonnet, E.; Arnaud, N.; Guivel, C.; Lagabrielle, Y.; Scalabrino, B.; Espinoza, F.

    2010-01-01

    The Chile Triple Junction is a natural laboratory to study the interactions between magmatism and tectonics during the subduction of an active spreading ridge beneath a continent. The MLBA plateau (Meseta del Lago Buenos Aires) is one of the Neogene alkali basaltic plateaus located in the back-arc region of the Andean Cordillera at the latitude of the current Chile Triple Junction. The genesis of MLBA can be related with successive opening of slabs windows beneath Patagonia: within the subducting Nazca Plate itself and between the Nazca and Antarctic plates. Detailed 40Ar/ 39Ar dating and geochemical analysis of bimodal magmatism from the western flank of the MLBA show major changes in the back-arc magmatism which occurred between 14.5 Ma and 12.5 Ma with the transition from calc-alkaline lavas (Cerro Plomo) to alkaline lavas (MLBA) in relation with slab window opening. In a second step, at 4-3 Ma, alkaline felsic intrusions were emplaced in the western flank of the MLBA coevally with the MLBA basalts with which they are genetically related. These late OIB-like alkaline to transitional basalts were generated by partial melting of the subslab asthenosphere of the subducting Nazca plate during the opening of the South Chile spreading ridge-related slab window. These basalts differentiated with small amounts of assimilation in shallow magma chambers emplaced along transtensional to extensional zones. The close association of bimodal magmatism with extensional tectonic features in the western MLBA is a strong support to the model of Patagonian collapse event proposed to have taken place between 5 and 3 Ma as a consequence of the presence of the asthenospheric window (SCR-1 segment of South Chile Ridge) below the MLBA area.

  3. Technical note on a track-pattern-based model for predicting seasonal tropical cyclone activity over the western North Pacific

    NASA Astrophysics Data System (ADS)

    Ho, Chang-Hoi; Kim, Joo-Hong; Kim, Hyeong-Seog; Choi, Woosuk; Lee, Min-Hee; Yoo, Hee-Dong; Kim, Tae-Ryong; Park, Sangwook

    2013-09-01

    Recently, the National Typhoon Center (NTC) at the Korea Meteorological Administration launched a track-pattern-based model that predicts the horizontal distribution of tropical cyclone (TC) track density from June to October. This model is the first approach to target seasonal TC track clusters covering the entire western North Pacific (WNP) basin, and may represent a milestone for seasonal TC forecasting, using a simple statistical method that can be applied at weather operation centers. In this note, we describe the procedure of the track-pattern-based model with brief technical background to provide practical information on the use and operation of the model. The model comprises three major steps. First, long-term data of WNP TC tracks reveal seven climatological track clusters. Second, the TC counts for each cluster are predicted using a hybrid statistical-dynamical method, using the seasonal prediction of large-scale environments. Third, the final forecast map of track density is constructed by merging the spatial probabilities of the seven clusters and applying necessary bias corrections. Although the model is developed to issue the seasonal forecast in mid-May, it can be applied to alternative dates and target seasons following the procedure described in this note. Work continues on establishing an automatic system for this model at the NTC.

  4. Impact of reclamation treatment on the biological activity of soils of the solonetz complex in Western Siberia

    NASA Astrophysics Data System (ADS)

    Berezin, L. V.; Khamova, O. F.; Paderina, E. V.; Gindemit, A. M.

    2014-11-01

    The abundance and activity of the soil microflora were studied in a field experiment with the use of green manure crops to assess the impact of reclamation measures on the biological activity of soils of the solonetz complex. The number of microorganisms in the plow soil horizon increased in the background of the green fallows as compared to the black ones. Coefficients of mineralization, immobilization, and transformation of organic compounds were calculated for different variants of the soil treatment. The value of the mineralization coefficient indicates the intense decomposition of the green manure that entered the soil. In the first year, peas were actively decomposed, while oats, in the second year (aftereffect). The activity of the soil enzymes (invertase, urease, and catalase) was determined. A close relationship between the catalase activity and the intensity of the microbiological processes in the soils was revealed.

  5. Identification of Geomorphic Signatures of Neotectonic Activity Using dem in the Precambrian Terrain of Western Ghats, India

    NASA Astrophysics Data System (ADS)

    Jayappa, K. S.; Markose, V. J.; Nagaraju, M.

    2012-07-01

    To assess the relative tectonic activity classes, five geomorphic indices such as stream-gradient index (SL), drainage basin asymmetry (Af), hypsometric integral (Hi), valley floor width - valley height ratio (Vf) and drainage basin shape (Bs) of ninety-four sub-basins of Valapattanam river basin have been analysed by applying the standard formulae. Relative tectonic activity classes (Iat) obtained by the average (S/n) of different classes of geomorphic indices have been classified into three groups. Group I shows high tectonic activity with values of S/n < 2; group II shows moderate tectonic activity with S/n > 2 to < 2.5; and group III shows low tectonic activity with values of S/n ≥ 2.5. Field evidences such as deep valleys, sudden changes in the river course and waterfalls at fault planes clearly agree with the values and classes of tectonic geomorphic indices.

  6. Cadmium regulates copper homoeostasis by inhibiting the activity of Mac1, a transcriptional activator of the copper regulon, in Saccharomyces cerevisiae.

    PubMed

    Heo, Dong-Hyuk; Baek, In-Joon; Kang, Hyun-Jun; Kim, Ji-Hyun; Chang, Miwha; Jeong, Mi-Young; Kim, Tae-Hyoung; Choi, Il-Dong; Yun, Cheol-Won

    2010-10-15

    Cadmium is a toxic metal and the mechanism of its toxicity has been studied in various model systems from bacteria to mammals. We employed Saccharomyces cerevisiae as a model system to study cadmium toxicity at the molecular level because it has been used to identify the molecular mechanisms of toxicity found in higher organisms. cDNA microarray and Northern blot analyses revealed that cadmium salts inhibited the expression of genes related to copper metabolism. Western blotting, Northern blotting and chromatin immunoprecipitation experiments indicated that CTR1 expression was inhibited at the transcriptional level through direct inhibition of the Mac1 transcriptional activator. The decreased expression of CTR1 results in cellular copper deficiency and inhibition of Fet3 activity, which eventually impairs iron uptake. In this way, cadmium exhibits a negative effect on both iron and copper homoeostasis.

  7. [Litter decomposition and lignocellulose enzyme activities of Actinothuidium hookeri and Cys- topteris montana in alpine timberline ecotone of western Sichuan, China].

    PubMed

    Chen, Ya-mei; He, Run-lian; Deng, Chang-chun; Yang, Wan-qin; Zhang, Jian; Yang, Lin; Liu, Yang

    2015-11-01

    The mass loss and lignocellulose enzyme activities of Actinothuidium hookeri residues and Cystopteris montana leaf litter in coniferous forest and timberline of western Sichuan, China were investigated. The results showed that both the mass loss rates of A. hookeri and C. Montana in timberline were higher than those in coniferous forest, while enzyme activities in timberline were lower than those in coniferous forest which was contrast with the hypothesis. The mass loss of two ground covers had significant differences in different seasons. The mass loss rate of A. hookeri in snow-covered season accounted for 69.8% and 83.0% of the whole year' s in timberline and coniferous forest, while that of C. montana in the growing season accounted for 82.6% and 83.4% of the whole year' s in timberline and coniferous forest, respectively. C. montana leaf litter decayed faster in the growing season, which was consistent with its higher cellulase activity in the growing season. The result illustrated that the enzymatic hydrolysis of cellulose and hemicellulose might be the main driving force for the early stage of litter decomposition. Multiple linear regression analysis showed that environmental factors and initial litter quality could explain 45.8%-85.1% variation of enzyme activity. The enzyme activities of A. hookeri and C. montana in the process of decomposition were mainly affected by the freeze-thaw cycle in snow-covered season. PMID:26915177

  8. Office-Based Physical Activity and Nutrition Intervention: Barriers, Enablers, and Preferred Strategies for Workplace Obesity Prevention, Perth, Western Australia, 2012

    PubMed Central

    Jancey, Jonine; Howat, Peter; Ledger, Melissa; Lee, Andy H.

    2013-01-01

    Introduction Workplace health promotion programs to prevent overweight and obesity in office-based employees should be evidence-based and comprehensive and should consider behavioral, social, organizational, and environmental factors. The objective of this study was to identify barriers to and enablers of physical activity and nutrition as well as intervention strategies for health promotion in office-based workplaces in the Perth, Western Australia, metropolitan area in 2012. Methods We conducted an online survey of 111 employees from 55 organizations. The online survey investigated demographics, individual and workplace characteristics, barriers and enablers, intervention-strategy preferences, and physical activity and nutrition behaviors. We used χ2 and Mann–Whitney U statistics to test for differences between age and sex groups for barriers and enablers, intervention-strategy preferences, and physical activity and nutrition behaviors. Stepwise multiple regression analysis determined factors that affect physical activity and nutrition behaviors. Results We identified several factors that affected physical activity and nutrition behaviors, including the most common barriers (“too tired” and “access to unhealthy food”) and enablers (“enjoy physical activity” and “nutrition knowledge”). Intervention-strategy preferences demonstrated employee support for health promotion in the workplace. Conclusion The findings provide useful insights into employees’ preferences for interventions; they can be used to develop comprehensive programs for evidence-based workplace health promotion that consider environmental and policy influences as well as the individual. PMID:24028834

  9. The impact of human activities on land use and land cover changes and environmental processes in the Gorce Mountains (Western Polish Carpathians) in the past 50 years.

    PubMed

    Bucała, Anna

    2014-06-01

    The role of human impact on the natural environment was studied in the Jaszcze and Jamne catchments in the Gorce Mountains (Western Polish Carpathians). Analysis of land use and land cover changes using GIS techniques and cartographic materials between 1954 and 2004 indicates an increase in forest area by 11.5% and 18.5%, respectively, at the expense of arable land and grassland areas. Agricultural abandonment often occurred on steep slopes (above 10°) with skeletal (shallow) soils at higher elevations (above 800 m a.s.l.). In addition, the density of dirt roads decreased from 6.97 km/km(2) in 1981 to 4.3 km/km(2) in 2008. In former agricultural areas, long expanses of terraces have either disappeared or experienced shallow mass movements. The statistical reports and questionnaire survey indicate reduced income from farm activities in this region. As a result of LULC changes and stream transformation, the Jaszcze and Jamne stream channels were shortened, straightened, and narrowed, with tendency to incision estimated at 1 cm per year over the past 40 years. The changes observed in the environment under human impact, accelerated following 1989, are representative of the Western Polish Carpathians. PMID:24565936

  10. [Recent progress of international harmonization of crude drugs and medicinal plants--activity of FHH (The Western Pacific Regional Forum for the Harmonization of Herbal Medicines)].

    PubMed

    Kawahara, Nobuo

    2011-03-01

    The Western Pacific Regional Forum for the Harmonization of Herbal Medicines (FHH) was established in 2002. The general proposed objective of the FHH is to promote public health by recognizing and developing standards and technical guidelines that aim to improve the quality, safety and efficacy of herbal medicines. At a sub-committee meeting of FHH nomenclature and standardization held in Tokyo, all the participants recognized the importance of comparing the descriptions of herbal medicines contained in member countries' pharmacopoeias or monograph standards as the first step in the harmonization of nomenclature and standardization. It was agreed to set up five expert working groups (EWG) to carry out the following specific tasks: 1) Nomenclature, 2) Testing Methods in Monographs, 3) List of Chemical Reference Standards (CRS) and Reference of Medicinal Plant Materials (RMPM), 4) List of Analytically Validated Methods, and 5) Information on General Tests. In this review, we report four topics of FHH activities from 2002-2009 as follows: 1) Comparative study on testing methods and specification values for crude drugs used in monographs among four Western Pacific regional countries (Japan, China, Korea and Vietnam), 2) Comparative study on TLC conditions for identification, chemical assay conditions for component quantification used in monographs among the four countries, 3) Comparative study on general testing methods for crude drugs among the four countries, 4) Comparative study on TLC identification for crude drugs used in monographs among the four countries considering harmonization and clean analysis.

  11. High Resolution Geophysical Survey of Western Long Island Sound Offshore New York: An Estuary Floor Shaped by Bottom Currents and Human Activity

    NASA Astrophysics Data System (ADS)

    Vargas, W.; Cormier, M.; McHugh, C.

    2009-05-01

    Western Long Island Sound near metropolitan New York averages 16m in water depth, with a few elongated depression up to 40 m deep. In June 2006, we surveyed the westernmost section of Long island Sound with the R/V HUGH SHARP. Analysis of the high-resolution multibeam bathymetric data collected during that survey reveals a series of sedimentary features that are consistent with a net westward direction of bottom currents. These features include: (1) Large sedimentary waves spaced ab out 100 m west of two km-scale features outcropping through the sediments; (2) Prominent sediment drifts or scour marks west of numerous shipwrecks and bouldery outcrops; (3) Series of subtle, sub-parallel sedimentary furrows aligned in a general EW direction along the north slope of the surveyed area (similar features have been documented in north- central Long Island Sound - (Poppe et al., J. Coastal Res. 2002). The lack of short wavelength sedimentary waves is consistent with the known muddy substrate and weak bottom currents (<10 cm/s) documented in western Long Island Sound. Fields of pockmarks affecting areas of gas-charged sediments may indicate localized, active venting of fluids and/or gas. The high-resolution bathymetry also highlights numerous anthropogenic disturbances such as pipelines, cables, shipwrecks, anchor drag marks, and dredge spoils.

  12. Chemical composition and biological activity of essential oils of Dracocephalum heterophyllum and Hyssopus officinalis from Western Himalaya

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The essential oils of two representatives of the Lamiaceae-family, Dracocephalum heterophyllum Benth. and Hyssopus officinalis L., are described for their antifungal, antibacterial and larvicidal as well as biting deterrent activities. Additionally, the essential oils’ chemical compositions, analyze...

  13. Chemical composition and biological activity of essential oils of Dracocephalum heterophyllum and Hyssopus officinalis from Western Himalaya.

    PubMed

    Stappen, Iris; Wanner, Jürgen; Tabanca, Nurhayat; Wedge, David E; Ali, Abbas; Kaul, Vijay K; Lal, Brij; Jaitak, Vikas; Gochev, Velizar K; Schmidt, Erich; Jirovetz, Leopold

    2015-01-01

    The essential oils of two representatives of the Lamiaceae, Dracocephalum heterophyllum Benth. and Hyssopus officinalis L., are described for their antifungal, antibacterial, larvicidal and inect biting deterrent activities. Additionally, the chemical compositions of the essential oils, analyzed by simultaneous GC-MS and GC-FID, and odor descriptions are given. The main components of H. officinalis oil were pinocarvone, cis-pinocamphone, and β-pinene. Citronellol was found as the main compound of D. heterophyllum essential oil. Antibacterial testing by agar dilution assay revealed greater activity of D. heterophyllum against Staphylococcus aureus compared with H. officinalis. D. heterophyllum essential oil also showed promising antifungal activity against Colletotrichum species and was more toxic to Aedes aegypti larvae in a larvicial bioassay. Both essential oils showed high activity in the biting deterrent bioassay.

  14. Chemical composition and biological activity of essential oils of Dracocephalum heterophyllum and Hyssopus officinalis from Western Himalaya.

    PubMed

    Stappen, Iris; Wanner, Jürgen; Tabanca, Nurhayat; Wedge, David E; Ali, Abbas; Kaul, Vijay K; Lal, Brij; Jaitak, Vikas; Gochev, Velizar K; Schmidt, Erich; Jirovetz, Leopold

    2015-01-01

    The essential oils of two representatives of the Lamiaceae, Dracocephalum heterophyllum Benth. and Hyssopus officinalis L., are described for their antifungal, antibacterial, larvicidal and inect biting deterrent activities. Additionally, the chemical compositions of the essential oils, analyzed by simultaneous GC-MS and GC-FID, and odor descriptions are given. The main components of H. officinalis oil were pinocarvone, cis-pinocamphone, and β-pinene. Citronellol was found as the main compound of D. heterophyllum essential oil. Antibacterial testing by agar dilution assay revealed greater activity of D. heterophyllum against Staphylococcus aureus compared with H. officinalis. D. heterophyllum essential oil also showed promising antifungal activity against Colletotrichum species and was more toxic to Aedes aegypti larvae in a larvicial bioassay. Both essential oils showed high activity in the biting deterrent bioassay. PMID:25920235

  15. 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

  16. What Contributes to the Activeness of Ethnic Minority Patients with Chronic Illnesses Seeking Allied Health Services? A Cross-Sectional Study in Rural Western China.

    PubMed

    Tang, Shangfeng; Dong, Dong; Ji, Lu; Fu, Hang; Feng, Zhanchun; Bishwajit, Ghose; He, Zhifei; Ming, Hui; Fu, Qian; Xian, Yue

    2015-09-15

    Actively seeking health services lies at the core of effective models of chronic disease self-management and contributes to promoting the utilization of allied health services (AHS). However, the use of AHS by ethnic minority Chinese, especially the elderly living in rural areas, has not received much attention. This study, therefore, aims to explore the association between personal characteristics and the activeness of ethnic minority patients with chronic diseases in rural areas of western China seeking AHS. A cross-sectional study was conducted to collect data on the socio-demographic and economic characteristics, health knowledge level and health communication channels of the sampled patients. A logistic regression model was used to examine the association of these predictors with the activeness of the surveyed patients in seeking AHS. A total of 1078 ethnic minorities over 45 years old who had chronic conditions were randomly selected from three western provinces in China and were interviewed in 2014. It is found that the New Cooperative Medical Scheme (NCMS) is the most salient predictor affecting the activeness of Chinese ethnic minorities in seeking AHS. The probability is 8.51 times greater for those insured with NCMS to actively seek AHS than those without (95% Confidence Interval (CI) 4.76-15.21; p < 0.001). Moreover, participants between 60 and 70 years old and those who have five to six household members are more likely to seek AHS compared with other social groups (Odds Ratio (OR) = 1.64, 95% CI 1.28-2.97, p = 0.007; OR = 1.95, 95% CI 1.15-2.36, p = 0.002). However, the activeness of patients seeking AHS is lower for those who have better household economic conditions. Besides socio-demographic predictors, the Chinese ethnic minorities' activeness in seeking AHS is clearly associated with the communication channels used for receiving health information, which include direct communication with doctors (OR = 5.18, 95% CI 3.58-7.50, p < 0.001) and

  17. [Measurement of antiganglioside autoantibodies by immunodot-blot assay: clinical importance in peripheral neuropathies].

    PubMed

    Caudie, C; Vial, C; Petiot, P; Bancel, J; Later, R; Gonnaud, P M

    1999-01-01

    We retrospectively evaluated measurement data and clinical relevance of autoantibodies to gangliosides in peripheral neuropathies (PN). The IgG and IgM antiganglioside autoantibodies were determined by our own immunodot-blot assay on membrane and by enzyme-linked immunosorbent assay (Elisa) in sera of 1,342 patients with peripheral neuropathies. Anti-GM1 and anti-GD1b autoantibodies formed a part of the normal autoantibody repertoire and were common place in 12% of normal subjects and in 14% of disease control groups. Polyclonal IgM antiganglioside autoantibodies were detected in chronic PN, polyclonal IgG antiganglioside autoantibodies were detected in acute PN. Polyclonal IgM anti-GM1 and anti-GD1b autoantibodies were detected in 35 patients out of 48 with treatable multifocal motor neuropathy with persistent conduction blocks. These autoantibodies well discriminated between suspected motor peripheral neuropathies and motor neuron diseases (sensitivity 73%, specificity 83%, positive predictive value 60%, negative predictive value 91%). Monoclonal IgM autoantibodies reacted strongly with gangliosides in 15 patients out of 77 with M-IgM neuropathy (19%). M-IgM autoantibodies differed in their fine specificities with different principal target antigens as demonstrated with cross-reactivity. Such findings provide further evidence for a relationship between neurological syndromes and antiganglioside antibody profiles and also suggest that different gangliosides could be principal target antigens such as GM1, GD1b, GT1b, GD1a or GM2. Polyclonal IgG anti-GM1 and anti-GD1b autoantibodies were detected in 21 patients out of 22 with acute motor axonal Guillain-Barré syndrome with antecedent of infection by Campylobacter jejuni, polyclonal IgG anti-GQ1b autoantibodies in 9 patients out of 10 with Miller-Fisher syndrome. Detection of antiganglioside autoantibodies by immunodot-blot assay which is simple and quick in testing a large panel of gangliosides has become very

  18. Effect of the permafrost thawing on the organic carbon and microbial activity in thermokarst lakes of Western Siberia: important source of carbon dioxide in the atmosphere

    NASA Astrophysics Data System (ADS)

    Shirokova, L. S.; Pokrovsky, O. S.; Kirpotin, S. N.; Dupre, B.

    2008-12-01

    Ongoing processes of the permafrost thawing in Western Siberia are likely to increase the surface of water reservoirs via forming so-called thermokarst lakes, mobilizing the organic carbon (OC) from the soil pool to the rivers and, finally, to the ocean, and thus modifying the fluxes of methane and CO2 to the atmosphere. In order to understand the mechanisms of carbon mobilization and biodegradation during permafrost thawing and to establish the link between the OC and microbial activity in forming thermokarstic lakes, we performed a comparative multidisciplinary study on the biogeochemistry of OC and metals in lakes located in the northern part of Western Siberia. About 10 lakes and small ponds of various size and age were sampled for dissolved and colloidal organic carbon and metals and total bacterial cell number. There is a systematic evolution of DOC, pH, trace elements and biological activity during successions of thermokarst lakes encountered in the present study. At the beginning of permafrost thawing at the scale of several meter size ponds, fast lixiviation of unaltered peat yields significant amount of OC, major and trace elements; the pH of these waters is between 3.5 and 4.0 and the conductivity is 20-30 μS. The intermediate stage of lake formation still preserve low pHs, high DOC and conductivity, even in relatively large, up to 1 km diameter but fast growing lakes. At these stages, there is no any productivity as phytoplankton cannot live in these acidic waters and bacterial mineralization intensity is around 0.3 mg C/L/day both in the surface and bottom horizons. Once the lake border is stabilized, there are no new "unaltered peat" sources and the biological processes start to consume the OC and nutrients. At this stage, there is still no production in the water column (< 0.01 mg C/L/day) but the bacterial mineralization intensity remains high, up to 0.3 mg C/L/day. At this final stage, the remaining part of the lake located in the centre of the

  19. Active Fault Deformation Along the South Boundary of the Western Transverse Ranges Province, Point Dume to the Northern Channel Islands, Southern California

    NASA Astrophysics Data System (ADS)

    Fisher, M. A.; Langenheim, V. E.; Sorlien, C. C.; Nicholson, C.; Sliter, R. W.

    2005-12-01

    The regional fault system forming the south boundary of the Western Transverse Ranges Province (WTRP) extends for about 200 km, from near the city of Los Angeles westward along the south flank of the Santa Monica Mountains and through the northern Channel Islands. Multichannel seismic-reflection data show that fault strands within the province-bounding system are active, and some have dip-slip displacements measured in kilometers. The left-oblique Dume fault is active and shows large displacement as far west as Sycamore knoll, but farther west, the fault tip and a superjacent fold are deeply buried. Thus during future earthquakes, the structural transition near the knoll could represent a boundary between earthquake-rupture segments. The east-west province-bounding fault system strikes at a high angle across and terminates the northwest-trending faults, basins and ridges of the California Continental Borderland. Borderland structures considered here form the western limit of intense middle Miocene oblique extension that accompanied rotation of the WTRP. The transition between extended and intact crust lies along the northwest-trending Santa Cruz-Catalina and Santa Rosa-Cortes Ridges. After the Miocene rotation, structures within these ridges became involved in regional transpression, such that northwestward along the Santa Cruz-Catalina Ridge, thrust faulting becomes increasingly more intense, and adjacent to the province boundary, thrust-faulted rocks completely override Miocene extensional structures. In contrast, rocks making up the Santa Rosa-Cortez Ridge are little deformed. The difference in deformation of the two ridges could result from a combination of: 1) eastward crustal thinning and consequent weakening that developed during the Miocene extension; 2) a difference in horizontal strain across the right-slip San Clemente fault near its termination at the WTRP boundary; 3) strain partitioning along this boundary; and 4) a contrast in bulk rheological

  20. Determining the Through-Going Active Fault Geometry of the Western North Anatolian Fault Through Stress Modeling

    NASA Astrophysics Data System (ADS)

    Karimi, B.; McQuarrie, N.

    2015-12-01

    The North Anatolian Fault (NAF) is a seismically active 1200 km long dextral strike-slip fault part of an east-west trending dextral shear zone (NAF system) between the Anatolian and Eurasian plates. This shear zone widens to the west, complicating potential earthquake rupture paths and highlighting the importance of understanding the geometry of active fault systems. West of the town of Bolu - the NAF bifurcates into the northern and southern strands, which converge and are linked through the Mudurnu Valley, then diverge to border the Marmara Sea. The westward continuation of these two fault traces is marked by further complexities in potential active fault geometry, particularly in the Marmara Sea (northern strand), and the Biga Peninsula (southern strand). We evaluate potential active fault geometries for both strands by comparing stress models of various fault geometries in these regions to a record of focal mechanisms and inferred paleostress from a lineament analysis. For the Marmara region, two of the three possible geometries matched the maximum horizontal stress (σH) orientations determined from a record of focal mechanisms; however, only one represented the northern and southern sidewalls associated with the principal zone of deformation of the developing Marmara basin. This suggests that it is the most likely representation of the active through-going fault geometry in the region. In the Biga Peninsula region, the active geometry of the southern strand has the southern component approaching and intersecting the northern component through a linking feature in a narrow topographic valley. This geometry was selected over two others as it overlaps the σH orientation determined from focal mechanism data and a lineament analysis. Additionally, this geometry does not develop a prominent mis-oriented NE-SW stress feature observed in the model results of the other two geometries, otherwise absent in the focal mechanism data or inferred from a lineament analysis.

  1. Rapid detection of intestinal pathogens in fecal samples by an improved reverse dot blot method

    PubMed Central

    Xing, Jian-Ming; Zhang, Su; Du, Ying; Bi, Dan; Yao, Li-Hui

    2009-01-01

    AIM: To develop a new, rapid and accurate reverse dot blot (RDB) method for the detection of intestinal pathogens in fecal samples. METHODS: The 12 intestinal pathogens tested were Salmonella spp., Brucella spp., Escherichia coli O157:H7, Clostridium botulinum, Bacillus cereus, Clostridium perfringens, Vibrio parahaemolyticus, Shigella spp., Yersinia enterocolitica, Vibrio cholerae, Listeria monocytogenes and Staphylococcus aureus. The two universal primers were designed to amplify two variable regions of bacterial 16S and 23S rDNA genes from all of the 12 bacterial species tested. Five hundred and forty fecal samples from the diarrhea patients were detected using the improved RDB assay. RESULTS: The methods could identify the 12 intestinal pathogens specifically, and the detection limit was as low as 103 CFUs. The consistent detection rate of the improved RDB assay compared with the traditional culture method was up to 88.75%. CONCLUSION: The hybridization results indicated that the improved RDB assay developed was a reliable method for the detection of intestinal pathogen in fecal samples. PMID:19469006

  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. 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

  4. Binding of 16S rRNA to chloroplast 30S ribosomal proteins blotted on nitrocellulose.

    PubMed

    Rozier, C; Mache, R

    1984-10-11

    Protein-RNA associations were studied by a method using proteins blotted on a nitrocellulose sheet. This method was assayed with Escherichia Coli 30S ribosomal components. In stringent conditions (300 mM NaCl or 20 degrees C) only 9 E. coli ribosomal proteins strongly bound to the 16S rRNA: S4, S5, S7, S9, S12, S13, S14, S19, S20. 8 of these proteins have been previously found to bind independently to the 16S rRNA. The same method was applied to determine protein-RNA interactions in spinach chloroplast 30S ribosomal subunits. A set of only 7 proteins was bound to chloroplast rRNA in stringent conditions: chloroplast S6, S10, S11, S14, S15, S17 and S22. They also bound to E. coli 16S rRNA. This set includes 4 chloroplast-synthesized proteins: S6, S11, S15 and S22. The core particles obtained after treatment by LiCl of chloroplast 30S ribosomal subunit contained 3 proteins (S6, S10 and S14) which are included in the set of 7 binding proteins. This set of proteins probably play a part in the early steps of the assembly of the chloroplast 30S ribosomal subunit.

  5. Reverse line blot macroarray for simultaneous detection and characterization of four biological warfare agents.

    PubMed

    Vanlalhmuaka; Thavachelvam, Kulanthaivel; Tuteja, Urmil; Sarika, Kumari; Nagendra, Suryanarayana; Kumar, Subodh

    2013-03-01

    The need for a rapid detection and characterization of biowarfare (BW) agents cannot be over emphasized. With diverse array of potential BW pathogen available presently, rapid identification of the pathogen is crucial, so that specific therapy and control measures can be initiated. We have developed a multiplex polymerase chain reaction based reverse line blot macroarray to simultaneously detect four pathogens of BW importance viz. Bacillus anthracis, Yersinia pestis, Brucella melitensis and Burkholderia pseudomallei. The multiplex PCR utilizes 14 pairs of primers targeting 18 specific markers. These markers include genes which are genus specific, species-specific chromosomal sequences and virulence markers of plasmid origin. The assay was evaluated on various human, environment and animal isolates. The assay w successful in simultaneous detection and characterization of isolates of the four pathogens on as a single platform with sensitivity ranging from 0.3 pg to 0.3 ng of genomic DNA. The assay was able to detect 5 × 10(2) cfu/ml for B. anthracis, 8 × 10(2) cfu/ml for Yersinia sp., 1.4 × 10(2) cfu/ml for B. melitensis and 4 × 10(2) cfu/ml for B. pseudomallei.

  6. 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

  7. Effects of food availability and climate on activity patterns of western black-crested gibbons in an isolated forest fragment in southern Yunnan, China.

    PubMed

    Ni, Qingyong; Xie, Meng; Grueter, Cyril C; Jiang, Xuelong; Xu, Huailiang; Yao, Yongfang; Zhang, Mingwang; Li, Yan; Yang, Jiandong

    2015-10-01

    The critically endangered western black-crested gibbon (Nomascus concolor) is distributed in isolated habitat fragments in northern Vietnam, northwestern Laos, and southwestern China. To assess the behavioral adaptation of this species to forest fragments and its response to seasonal variation in food availability and climate, we present activity patterns of a group inhabiting an isolated forest based on two year-long studies in southern Yunnan, China. Annually, the gibbons spent nearly half of their active time resting, followed by moving and feeding. In both study periods, the time allocated to activities varied significantly between months, and was affected by food availability and climate factors. The group delayed retirement when tree fruit was abundant, and they decreased time spent moving and playing during periods of low fruit availability. In the cold months, the gibbons decreased time spent moving, and they decreased active time and resting time when rainfall was high. The results suggest that the group may seek to maximize net energy intake like energy maximizers when high quality food is most available, and adopt an energy-conserving strategy during periods of lower food availability and temperature. The gibbons showed similar diurnal variation in activity patterns to a group inhabiting a continuous forest at Dazhaizi, Mt. Wuliang, central Yunnan. However, they had a longer active period, and devoted more time to resting but less time to feeding. The individuals also spent lower percentages of time engaged in social behavior than those at Dazhaizi. These differences may be due to their smaller home range and unusual group composition caused by habitat fragmentation. PMID:26275664

  8. Emergence and seasonal activity of the entomophagous rove beetle Aleochara bilineata (Coleoptera: Staphylinidae) in canola in Western Canada.

    PubMed

    Broatch, J S; Dosdall, L M; Yang, R-C; Harker, K N; Clayton, G W

    2008-12-01

    Aleochara bilineata Gyllenhal (Coleoptera: Staphylinidae) is an important natural enemy of root maggots (Delia spp.) (Diptera: Anthomyiidae), which are serious pests of brassicaceous crops in North America and Europe. Adults of A. bilineata feed on eggs and larvae of root maggots, and A. bilineata larvae parasitize Delia spp. pupae. Emergence and seasonal activity patterns of A. bilineata were investigated during 2003-2005 in canola (Brassica rapa L. and Brassica napus L.) in central Alberta, Canada, in relation to degree-day (DD) accumulations and Julian date. Captures of A. bilineata adults from pitfall traps within emergence cages situated over canola stubble from the previous year indicated that approximately 428, 493, and 455 DD (soil base 5.57 degrees C) and 187, 189, and 180 Julian days were required for 50% emergence in 2003, 2004, and 2005, respectively (3-yr mean = 185.1 +/- 2.8 Julian days [SEM]). Captures of A. bilineata adults from pitfall traps placed in current canola crops determined that 50% levels of activity density required 379 DD and 180 Julian days in 2004. A logistic model that described the relationship of degree-days and Julian days with emergence of adult beetles was appraised, and good correspondence was evident between predicted and observed cumulative emergence patterns. Emergence and seasonal activity periods of A. bilineata in canola were well synchronized with occurrence of preimaginal life stages of its principal hosts, Delia radicum (L.) and Delia platura Meigen, with beetle emergence beginning shortly after the onset of root maggot oviposition.

  9. Chemical composition and antimicrobial activity of the essential oil of Plectranthus mollis (Lamiaceae) from Western Ghats Region, Karnataka, India.

    PubMed

    Joshi, Rajesh K

    2014-06-01

    Plectranthus is a large and widespread genus with a diversity of ethnobotanical uses. In traditional medicine P. mollis has been used against snakebites, respiratory stimulant and vasoconstrictor, cardiac depressant, cure for haemorrahage, treatment of mental retardation and rheumatism. P. mollis is reported to exhibit relaxant activity on smooth and skeletal muscles, and has cytotoxic and anti-tumour promoting activity, and can be used in the treatment of cancer. The aim of the present study was to identify chemical composition of the essential oil of P. mollis and to evaluate antimicrobial efficacy of the oil. The essential oil of the flowering aerial parts of P. mollis was obtained by hydro-distillation and analyzed by gas chromatography equipped with a flame ionization detector (GC-FID) and gas chromatography coupled with mass spectrometry (GC/MS). Twenty-seven compounds were identified, which comprised 98.6% of the total constituents. The main compound was identified as fenchone (32.3%), followed by alpha-humulene (17.3%), piperitenone oxide (8.5%), cis-piperitone oxide (6.0%) and E-beta-farnesene (5.9%). The oil was found rich in oxygenated monoterpenes type constituents (52.0%), followed by sesquiterpene hydrocarbons (40.2%), oxygenated sesquiterpenes (4.9%), and monoterpene hydrocarbons (1.5%). Antimicrobial activity of the essential oil of P. mollis was tested against six Gram-positive and eight Gram-negative bacteria, and three fungi, by using the tube dilution method. The oil was active against the tested Gram-positive and Gram-negative bacteria, and fungi at a concentration range of 0.065 +/- 0.008-0.937 +/- 0.139 mg/mL, 0.468 +/- 0.069-3.333 +/- 0.527 mg/mL and 0.117 +/- 0.017-0.338 +/- 0.062 mg/mL, respectively. The present study revealed that the oil constituents somehow were qualitatively similar and quantitatively different than earlier reports from different parts of the world. The essential oil of P. mollis has found to be antimicrobial activity

  10. Chemical composition and antimicrobial activity of the essential oil of Plectranthus mollis (Lamiaceae) from Western Ghats Region, Karnataka, India.

    PubMed

    Joshi, Rajesh K

    2014-06-01

    Plectranthus is a large and widespread genus with a diversity of ethnobotanical uses. In traditional medicine P. mollis has been used against snakebites, respiratory stimulant and vasoconstrictor, cardiac depressant, cure for haemorrahage, treatment of mental retardation and rheumatism. P. mollis is reported to exhibit relaxant activity on smooth and skeletal muscles, and has cytotoxic and anti-tumour promoting activity, and can be used in the treatment of cancer. The aim of the present study was to identify chemical composition of the essential oil of P. mollis and to evaluate antimicrobial efficacy of the oil. The essential oil of the flowering aerial parts of P. mollis was obtained by hydro-distillation and analyzed by gas chromatography equipped with a flame ionization detector (GC-FID) and gas chromatography coupled with mass spectrometry (GC/MS). Twenty-seven compounds were identified, which comprised 98.6% of the total constituents. The main compound was identified as fenchone (32.3%), followed by alpha-humulene (17.3%), piperitenone oxide (8.5%), cis-piperitone oxide (6.0%) and E-beta-farnesene (5.9%). The oil was found rich in oxygenated monoterpenes type constituents (52.0%), followed by sesquiterpene hydrocarbons (40.2%), oxygenated sesquiterpenes (4.9%), and monoterpene hydrocarbons (1.5%). Antimicrobial activity of the essential oil of P. mollis was tested against six Gram-positive and eight Gram-negative bacteria, and three fungi, by using the tube dilution method. The oil was active against the tested Gram-positive and Gram-negative bacteria, and fungi at a concentration range of 0.065 +/- 0.008-0.937 +/- 0.139 mg/mL, 0.468 +/- 0.069-3.333 +/- 0.527 mg/mL and 0.117 +/- 0.017-0.338 +/- 0.062 mg/mL, respectively. The present study revealed that the oil constituents somehow were qualitatively similar and quantitatively different than earlier reports from different parts of the world. The essential oil of P. mollis has found to be antimicrobial activity

  11. Distribution and relative activity of matrix metalloproteinase-2 in human coronal dentin

    PubMed Central

    Boushell, Lee W; Kaku, Masaru; Mochida, Yoshiyuki; Yamauchi, Mitsuo

    2011-01-01

    The presence of matrix metalloproteinase-2 (MMP-2) in dentin has been reported, but its distribution and activity level in mature human coronal dentin are not well understood. The purpose of this study was to determine the MMP-2 distribution and relative activity in demineralized dentin. Crowns of twenty eight human molars were sectioned into inner (ID), middle (MD), and outer dentin (OD) regions and demineralized. MMP-2 was extracted with 0.33 mol·L−1 EDTA/2 mol·L−1 guanidine-HCl, pH 7.4, and MMP-2 concentration was estimated with enzyme-linked immunoabsorbant assay (ELISA). Further characterization was accomplished by Western blotting analysis and gelatin zymography. The mean concentrations of MMP-2 per mg dentin protein in the dentin regions were significantly different (P=0.043): 0.9 ng (ID), 0.4 ng (MD), and 2.2 ng (OD), respectively. The pattern of MMP-2 concentration was OD>ID>MD. Western blotting analysis detected ∼66 and ∼72 kDa immunopositive proteins corresponding to pro- and mature MMP-2, respectively, in the ID and MD, and a ∼66 kDa protein in the OD. Gelatinolytic activity consistent with MMP-2 was detected in all regions. Interestingly, the pattern of levels of Western blot immunodetection and gelatinolytic activity was MD>ID>OD. The concentration of MMP-2 in human coronal dentin was highest in the region of dentin that contains the dentinoenamel junction and least in the middle region of dentin. However, levels of Western blot immunodetection and gelatinolytic activity did not correlate with the estimated regional concentrations of MMP-2, potentially indicating region specific protein interactions. PMID:22010577

  12. 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

  13. 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

  14. 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.

  15. 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.

  16. 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

  17. Performance of commercial reverse line blot assays for human papillomavirus genotyping.

    PubMed

    Steinau, Martin; Onyekwuluje, Juanita M; Scarbrough, Mariela Z; Unger, Elizabeth R; Dillner, Joakim; Zhou, Tiequn

    2012-05-01

    The performance of three line blot assays (LBAs), the Linear Array HPV genotyping assay (LA) (Roche Diagnostics), INNO-LiPA HPV Genotyping Extra (LiPA) (Innogenetics), and the reverse hybridization assay (RH) (Qiagen), was evaluated using quantitated whole genomic human papillomavirus (HPV) plasmids (types 6, 11, 16, 18, 31, 33, 35, 39, 51, 52, 56, 58, 59, and 68b) as well as epidemiologic samples. In a plasmid titration series, LiPA and RH did not detect 50 international units (IU) of HPV type 18 (HPV18) in the presence of 5 × 10(4) IU or more of HPV16. HPV DNA (1 to 6 types) in the plasmid challenges at 50 IU or genome equivalents (GE) were identified with an accuracy of 99.9% by LA, 97.3% by LiPA, and 95.4% by RH, with positive reproducibility of 99.8% (kappa = 0.992), 88.2% (kappa = 0.928), and 88.1% (kappa = 0.926), respectively. Two instances of mistyping occurred with LiPA. Of the 120 epidemiologic samples, 76 were positive for high-risk types by LA, 90 by LiPA, and 69 by RH, with a positive reproducibility of 87.3% (kappa = 0.925), 83.9% (kappa = 0.899), and 90.2% (kappa = 0.942), respectively. Although the assays had good concordance in the clinical samples, the greater accuracy and specificity in the plasmid panel suggest that LA has an advantage for internationally comparable genotyping studies.

  18. Impacts of decaying eastern and central Pacific El Niños on tropical cyclone activities over the western North Pacific in summer

    NASA Astrophysics Data System (ADS)

    Yang, Yuxing; Xie, Ruihuang; Wang, Faming; Huang, Fei

    2016-04-01

    We investigate the influences of the decaying eastern Pacific El Niño (EP - El Niño) and central Pacific El Niño (CP - El Niño) on tropical cyclone (TC) activities in the western North Pacific (WNP) during July, August and September (JAS). During this period, TC geneses and tracks are reduced in the central and eastern WNP. However, TC tracks reaching the Philippines increase, and more TC geneses appear west of 145°E during EP - El Niño. During CP - El Niño, tracks reaching the South China Sea (SCS) and southeast coast of China increase, and positive anomalies of TC genesis are found in the southern part of the central WNP and southern SCS. It is possible that the different variation of the anomalous anticyclone over east of the Philippines in the WNP induced by El Niños are instrumental to different TC variations in the two types of decaying El Niños during JAS. Compared with EP - El Niño, strengthening and northward expansion of the anomalous anticyclone during CP - El Niño cause a westward shift of the western Pacific subtropical high in summer, which is responsible for more westward TC tracks over the SCS and southeast coast of China. This northward expansion can cause the center of suppressed TC geneses in the central WNP to migrate further north during CP - El Niño. A decreased magnitude of vertical shear dominates the southern part of the central WNP and southern SCS, which enhances TC formation in these regions during CP - El Niño.

  19. Impacts of decaying eastern and central Pacific El Niños on tropical cyclone activities over the western North Pacific in summer

    NASA Astrophysics Data System (ADS)

    Yang, Yuxing; Xie, Ruihuang; Wang, Faming; Huang, Fei

    2016-07-01

    We investigate the influences of the decaying eastern Pacific El Niño (EP-El Niño) and central Pacific El Niño (CP-El Niño) on tropical cyclone (TC) activities in the western North Pacific (WNP) during July, August, and September (JAS). During this period, TC geneses and tracks are reduced in the central and eastern WNP. However, TC tracks reaching the Philippines increase, and more TC geneses appear west of 145°E during EP-El Niño. During CP-El Niño, tracks reaching the South China Sea (SCS) and southeast coast of China increase, and positive anomalies of TC genesis are found in the southern part of the central WNP and southern SCS. It is possible that the different variations of the anomalous anticyclone over east of the Philippines in the WNP induced by El Niños are instrumental to the different TC variations in the two types of decaying El Niños during JAS. Compared with EP-El Niño, strengthening and northward expansion of the anomalous anticyclone during CP-El Niño cause a westward shift of the western Pacific subtropical high in summer, which is responsible for more westward TC tracks over the SCS and southeast coast of China. This northward expansion can cause the center of suppressed TC geneses in the central WNP to migrate further north during CP-El Niño. A decreased magnitude of vertical shear dominates the southern part of the central WNP and southern SCS, which enhances TC formation in these regions during CP-El Niño.

  20. Reprint of “Zooplankton biomass and electron transport system activity around the Balearic Islands (western Mediterranean)”

    NASA Astrophysics Data System (ADS)

    Herrera, A.; Gómez, M.; Packard, T. T.; Fernández de Puelles, M. L.

    2014-10-01

    Measuring electron transport system (ETS) activity in zooplankton provides an index of respiration, theoretically, the potential respiration rate. We apply the ETS technique to estimate potential respiration and carbon demand from the zooplankton community in the upper 200 m of the water column near the Balearic Islands. The investigation was focused on two areas with different oceanographic conditions: the Balearic and Algerian subbasins. It compared the biomass, potential respiration and specific potential respiration of different size fractions (53-200, 200-500, > 500 μm) in both areas. In these regions the largest contribution to respiration was found in the larger sizes. The specific respiration (per unit biomass) was greater in smaller fractions, indicating that they have a more active metabolism. Both biomass and potential respiration increased in the Algerian subbasin and for both regions biomass and potential respiration were greater in shallow waters over the continental shelf (< 200 m). Using Kleiber's law as a tool to investigate the relationships between these two variables, we found that the exponential relation coefficient (b) was less than 0.75, indicating that the respiration was depressed (shifted down). In cultures and in eutrophic ocean waters (upwelling areas) b normally is greater than 0.75, consequently we intuit that the low value of b over the Balearic and Algerian subbasins indicates that the zooplankton is not well fed and that they are living under oligotrophic stress.

  1. Diel changes in food and feeding activity of sciaenid fishes from the South-western Atlantic, Brazil.

    PubMed

    Soares, L S; Vazzoler, A E

    2001-05-01

    Food habits and daily feeding activity of fish populations are important ecological variables for understanding their role in the ecosystem. Diel changes in feeding activity and dietary composition of juvenile whitemouth croaker Micropogonias furnieri, shortfin corvina Isopisthus parvipinnis, shorthead drum Larimus breviceps and of juvenile and adult banded croaker Paralonchurus brasiliensis were investigated from samples taken over a 24 hours period from 4 to 5 September 1987 on the continental shelf off South-eastern Brazil. Whitemouth croaker and banded croaker had a benthic diet based on polychaetes and some caridean shrimps and showed no diel feeding pattern. Shortfin corvina and shorthead drum fed on crustaceans and teleostean fish, the former species primarily on pelagic sergestid shrimps and benthic caridean shrimps, and the latter mainly on pelagic sergestid shrimps, mysidacean and benthic caridean shrimps. Shortfin corvina is primarily a diurnal feeder, but preyed on some items only at night. Shorthead drum is a night feeder, with minimum stomach fullness values between dawn and daytime, and maximum values between dusk and night time.

  2. Chemical profile, antiproliferative and antioxidant activities of rhizome oil of Zingiber anamalayanum from Western Ghats in India.

    PubMed

    Salim, Mohamed; Kabeer, T K Ahmedul; Nair, S Ajikumaran; Dan, Mathew; Sabu, M; Baby, Sabulal

    2016-09-01

    Volatile oil from fresh rhizomes of Zingiber anamalayanum was isolated by hydrodistillation and characterised by GC-FID and GC-MS. Twenty-one out of 24 constituents comprising 99.47% of the oil were identified. Major components in Z. anamalayanum rhizome oil were δ-2-carene (52.83%), camphene (9.83%), endo-fenchol (9.42%), iso-dihydrocarveol (6.44%) and cis-p-mentha-2,8-dien-1-ol (5.19%). Monoterpene hydrocarbons in the rhizome oil were 65.81%, followed by oxygenated monoterpenes (23.78%) and sesquiterpene hydrocarbons (9.87%). Physical parameters of rhizome oil were [Formula: see text] 1.4031, [Formula: see text] - 16.097(o) (c = 1, CHCl3) and [Formula: see text] 0.9202. Z. anamalayanum rhizome oil showed significant anti-Dalton's Lymphoma Ascitic activity. PMID:26456637

  3. Chemical profile, antiproliferative and antioxidant activities of rhizome oil of Zingiber anamalayanum from Western Ghats in India.

    PubMed

    Salim, Mohamed; Kabeer, T K Ahmedul; Nair, S Ajikumaran; Dan, Mathew; Sabu, M; Baby, Sabulal

    2016-09-01

    Volatile oil from fresh rhizomes of Zingiber anamalayanum was isolated by hydrodistillation and characterised by GC-FID and GC-MS. Twenty-one out of 24 constituents comprising 99.47% of the oil were identified. Major components in Z. anamalayanum rhizome oil were δ-2-carene (52.83%), camphene (9.83%), endo-fenchol (9.42%), iso-dihydrocarveol (6.44%) and cis-p-mentha-2,8-dien-1-ol (5.19%). Monoterpene hydrocarbons in the rhizome oil were 65.81%, followed by oxygenated monoterpenes (23.78%) and sesquiterpene hydrocarbons (9.87%). Physical parameters of rhizome oil were [Formula: see text] 1.4031, [Formula: see text] - 16.097(o) (c = 1, CHCl3) and [Formula: see text] 0.9202. Z. anamalayanum rhizome oil showed significant anti-Dalton's Lymphoma Ascitic activity.

  4. Track-pattern-based seasonal prediction model for intense tropical cyclone activities over the North Atlantic and the western North Pacific basins

    NASA Astrophysics Data System (ADS)

    Choi, W.; Ho, C. H.

    2015-12-01

    Intense tropical cyclones (TCs) accompanying heavy rainfall and destructive wind gusts sometimes cause incredible socio-economic damages in the regions near their landfall. This study aims to analyze intense TC activities in the North Atlantic (NA) and the western North Pacific (WNP) basins and develop their track propensity seasonal prediction model. Considering that the number of TCs in the NA basin is much smaller than that in the WNP basin, different intensity criteria are used; category 1 and above for NA and category 3 and above for WNP based on Saffir-Simpson hurricane wind scale. By using a fuzzy clustering method, intense TC tracks in the NA and the WNP basins are classified into two and three representative patterns, respectively. Each pattern shows empirical relationships with climate variabilities such as sea surface temperature distribution associated with El Niño/La Niña or Atlantic Meridional Mode, Pacific decadal oscillation, upper and low level zonal wind, and strength of subtropical high. The hybrid statistical-dynamical method has been used to develop the seasonal prediction model for each pattern based on statistical relationships between the intense TC activity and seasonal averaged key predictors. The model performance is statistically assessed by cross validation for the training period (1982-2013) and has been applied for the 2014 and 2015 prediction. This study suggests applicability of this model to real prediction work and provide bridgehead of attempt for intense TC prediction.

  5. Direct and indirect effects of petroleum production activities on the western fence lizard (Sceloporus occidentalis) as a surrogate for the dunes sagebrush lizard (Sceloporus arenicolus).

    PubMed

    Weir, Scott M; Knox, Ami; Talent, Larry G; Anderson, Todd A; Salice, Christopher J

    2016-05-01

    The dunes sagebrush lizard (Sceloporus arenicolus) is a habitat specialist of conservation concern limited to shin oak sand dune systems of New Mexico and Texas (USA). Because much of the dunes sagebrush lizard's habitat occurs in areas of high oil and gas production, there may be direct and indirect effects of these activities. The congeneric Western fence lizard (Sceloporus occidentalis) was used as a surrogate species to determine direct effects of 2 contaminants associated with oil and gas drilling activities in the Permian Basin (NM and TX, USA): herbicide formulations (Krovar and Quest) and hydrogen sulfide gas (H2S). Lizards were exposed to 2 concentrations of H2 S (30 ppm or 90 ppm) and herbicide formulations (1× or 2× label application rate) representing high-end exposure scenarios. Sublethal behavioral endpoints were evaluated, including sprint speed and time to prey detection and capture. Neither H2S nor herbicide formulations caused significant behavioral effects compared to controls. To understand potential indirect effects of oil and gas drilling on the prey base, terrestrial invertebrate biomass and order diversity were quantified at impacted sites to compare with nonimpacted sites. A significant decrease in biomass was found at impacted sites, but no significant effects on diversity. The results suggest little risk from direct toxic effects, but the potential for indirect effects should be further explored. PMID:26456391

  6. Effect of lead (Pb) exposure on the activity of superoxide dismutase and catalase in battery manufacturing workers (BMW) of Western Maharashtra (India) with reference to heme biosynthesis.

    PubMed

    Patil, Arun J; Bhagwat, Vinod R; Patil, Jyotsna A; Dongre, Nilima N; Ambekar, Jeevan G; Jailkhani, Rama; Das, Kusal K

    2006-12-01

    The aim of this study was to estimate the activity of superoxide dismutase (SOD) and catalase in erythrocytes and malondialdehyde (MDA) in plasma of battery manufacturing workers (BMW) of Western Maharashtra (India) who were occupationally exposed to lead (Pb) over a long period of time (about 15 years). This study was also aimed to determine the Pb intoxication resulted in a disturbance of heme biosynthesis in BMW group. The blood Pb level of BMW group (n = 28) was found to be in the range of 25.8 - 78.0 microg/dL (mean + SD, 53.63 + 16.98) whereas in Pb unexposed control group (n = 35) the range was 2.8 - 22.0 microg/dL (mean + SD, 12.52 + 4.08). The blood level (Pb-B) and urinary lead level (Pb-U) were significantly increased in BMW group as compared to unexposed control. Though activated d- aminolevulinic acid dehydratase (ALAD) activities in BMW group did not show any significant change when compared to control group but activated / non activated erythrocyte - ALAD activities in BMW group showed a significant increase. Erythrocyte- zinc protoporphyrin (ZPP), urinary daminolevulinic acid (ALA-U) and porphobilinogen (PBG-U) of BMW groups elevated significantly as compared to control. A positive correlation (r = 0.66, p < 0.001) between Pb-B and ALA-U were found in BMW group but no such significant correlation (r = 0.02, p> 1.0) were observed in control group. Hematological study revealed a significant decrease of hemoglobin concentration, packed cell volume (%) and other blood indices and a significant increase of total leucocytes count in BMW group in comparison to control group. The serum MDA content was significantly increased (p < 0.001) and the activities of antioxidant enzymes such as erythrocyte- SOD (p < 0.001) and erythrocytecatalase (p < 0.001) were significantly reduced in BMW group as compared to control group. A positive correlation (r = 0.45, p < 0.02) between Pb-B and serum MDA level was observed in BMW group (Pb-B range 25.8 - 78.0 microg / d

  7. Activation of the Nrf2 Cell Defense Pathway by Ancient Foods: Disease Prevention by Important Molecules and Microbes Lost from the Modern Western Diet.

    PubMed

    Senger, Donald R; Li, Dan; Jaminet, Shou-Ching; Cao, Shugeng

    2016-01-01

    The Nrf2 (NFE2L2) cell defense pathway protects against oxidative stress and disorders including cancer and neurodegeneration. Although activated modestly by oxidative stress alone, robust activation of the Nrf2 defense mechanism requires the additional presence of co-factors that facilitate electron exchange. Various molecules exhibit this co-factor function, including sulforaphane from cruciferous vegetables. However, natural co-factors that are potent and widely available from dietary sources have not been identified previously. The objectives of this study were to investigate support of the Nrf2 cell defense pathway by the alkyl catechols: 4-methylcatechol, 4-vinylcatechol, and 4-ethylcatechol. These small electrochemicals are naturally available from numerous sources but have not received attention. Findings reported here illustrate that these compounds are indeed potent co-factors for activation of the Nrf2 pathway both in vitro and in vivo. Each strongly supports expression of Nrf2 target genes in a variety of human cell types; and, in addition, 4-ethylcatechol is orally active in mice. Furthermore, findings reported here identify important and previously unrecognized sources of these compounds, arising from biotransformation of common plant compounds by lactobacilli that express phenolic acid decarboxylase. Thus, for example, Lactobacillus plantarum, Lactobacillus brevis, and Lactobacillus collinoides, which are consumed from a diet rich in traditionally fermented foods and beverages, convert common phenolic acids found in fruits and vegetables to 4-vinylcatechol and/or 4-ethylcatechol. In addition, all of the alkyl catechols are found in wood smoke that was used widely for food preservation. Thus, the potentially numerous sources of alkyl catechols in traditional foods suggest that these co-factors were common in ancient diets. However, with radical changes in food preservation, alkyl catechols have been lost from modern foods. The absence of alkyl

  8. Activation of the Nrf2 Cell Defense Pathway by Ancient Foods: Disease Prevention by Important Molecules and Microbes Lost from the Modern Western Diet.

    PubMed

    Senger, Donald R; Li, Dan; Jaminet, Shou-Ching; Cao, Shugeng

    2016-01-01

    The Nrf2 (NFE2L2) cell defense pathway protects against oxidative stress and disorders including cancer and neurodegeneration. Although activated modestly by oxidative stress alone, robust activation of the Nrf2 defense mechanism requires the additional presence of co-factors that facilitate electron exchange. Various molecules exhibit this co-factor function, including sulforaphane from cruciferous vegetables. However, natural co-factors that are potent and widely available from dietary sources have not been identified previously. The objectives of this study were to investigate support of the Nrf2 cell defense pathway by the alkyl catechols: 4-methylcatechol, 4-vinylcatechol, and 4-ethylcatechol. These small electrochemicals are naturally available from numerous sources but have not received attention. Findings reported here illustrate that these compounds are indeed potent co-factors for activation of the Nrf2 pathway both in vitro and in vivo. Each strongly supports expression of Nrf2 target genes in a variety of human cell types; and, in addition, 4-ethylcatechol is orally active in mice. Furthermore, findings reported here identify important and previously unrecognized sources of these compounds, arising from biotransformation of common plant compounds by lactobacilli that express phenolic acid decarboxylase. Thus, for example, Lactobacillus plantarum, Lactobacillus brevis, and Lactobacillus collinoides, which are consumed from a diet rich in traditionally fermented foods and beverages, convert common phenolic acids found in fruits and vegetables to 4-vinylcatechol and/or 4-ethylcatechol. In addition, all of the alkyl catechols are found in wood smoke that was used widely for food preservation. Thus, the potentially numerous sources of alkyl catechols in traditional foods suggest that these co-factors were common in ancient diets. However, with radical changes in food preservation, alkyl catechols have been lost from modern foods. The absence of alkyl

  9. Activation of the Nrf2 Cell Defense Pathway by Ancient Foods: Disease Prevention by Important Molecules and Microbes Lost from the Modern Western Diet

    PubMed Central

    Senger, Donald R.; Li, Dan; Jaminet, Shou-Ching; Cao, Shugeng

    2016-01-01

    The Nrf2 (NFE2L2) cell defense pathway protects against oxidative stress and disorders including cancer and neurodegeneration. Although activated modestly by oxidative stress alone, robust activation of the Nrf2 defense mechanism requires the additional presence of co-factors that facilitate electron exchange. Various molecules exhibit this co-factor function, including sulforaphane from cruciferous vegetables. However, natural co-factors that are potent and widely available from dietary sources have not been identified previously. The objectives of this study were to investigate support of the Nrf2 cell defense pathway by the alkyl catechols: 4-methylcatechol, 4-vinylcatechol, and 4-ethylcatechol. These small electrochemicals are naturally available from numerous sources but have not received attention. Findings reported here illustrate that these compounds are indeed potent co-factors for activation of the Nrf2 pathway both in vitro and in vivo. Each strongly supports expression of Nrf2 target genes in a variety of human cell types; and, in addition, 4-ethylcatechol is orally active in mice. Furthermore, findings reported here identify important and previously unrecognized sources of these compounds, arising from biotransformation of common plant compounds by lactobacilli that express phenolic acid decarboxylase. Thus, for example, Lactobacillus plantarum, Lactobacillus brevis, and Lactobacillus collinoides, which are consumed from a diet rich in traditionally fermented foods and beverages, convert common phenolic acids found in fruits and vegetables to 4-vinylcatechol and/or 4-ethylcatechol. In addition, all of the alkyl catechols are found in wood smoke that was used widely for food preservation. Thus, the potentially numerous sources of alkyl catechols in traditional foods suggest that these co-factors were common in ancient diets. However, with radical changes in food preservation, alkyl catechols have been lost from modern foods. The absence of alkyl

  10. Synthesis of the quaternary in the almeria littoral neotectonic activity and its morphologic features, western betics, Spain

    NASA Astrophysics Data System (ADS)

    Goy, J. L.; Zazo, C.

    1986-11-01

    A sequence of marine and continental Quaternary episodes is established, as are their interrelationships. The zone's tectonics have, in most cases, remained active to the present and control the factors All the Tyrrhenian levels containing Strombus bubonius are affected by the many faults that cross the lateral fringe and continue onto the shelf. Among these are: the Loma del Viento Fault, (N120°E. a normal fault with certain leftward characteristics): the El Alquian faults (N140°, 160°E, which coincide, according to several authors (Bousquet, 1979), with the surface feature of a great dextral deep accident); and the Carboneras Fault, or the Serrata Fault, (N40°, 45°E, left strike-slip fault). Given the varying behaviour of the shoreline, we have chosen to divide it into four sectors: Campo de Dalias, El-Alquian-Cabo de Gata, Mojacar—Garrucha. and Campo de Nijar-Serrata. Due to its geographical location, no marine-continental relationships exist in the latter. A chart has been made in an attempt to synthesize the most significant stratigraphic, genetic and tectonic characteristics of each sector. Lastly, the influence of tectonics on the distribution and heights of the Quaternary shorelines is clearly shown in a chart of the transgressive maxima of the Tyrrhenian episodes.

  11. Seasonal variations in active dispersal of natural populations of Triatoma infestans in rural north-western Argentina

    PubMed Central

    VAZQUEZ-PROKOPEC, G. M.; CEBALLOS, L.A.; MARCET, P. L.; CECERE, M. C.; CARDINAL, M. V.; KITRON, U.; GÜRTLER, R. E.

    2007-01-01

    The flight dispersal of Triatoma infestans Klug (Hemiptera: Reduviidae) is one of the main mechanisms determining community re-infestation after control interventions. An empirical model of flight initiation coupled with data from a longitudinal study predicted that the flight dispersal of T. infestans would peak in summer. To test this prediction, longitudinal light trap collections were conducted during 3-8 nights in March (late summer), July (winter) and November (spring) 2003, and in March 2004 in a rural community in north-west Argentina. Following each light-trapping collection date, all peridomestic sites around light traps were inspected to assess the relative abundance and nutritional status of T. infestans at each site. A total of 21 adult and five nymph T. infestans, six Triatoma guasayana Wygodzinsky & Abalos, and nine Triatoma garciabesi Carcavallo et al. were collected in 96 light-trapping nights, whereas 696 T. infestans were collected from the peridomestic sites that surrounded the light traps. The arrival of T. infestans in the light traps occurred in 64% of catch stations and peaked in the summer surveys (10-14 bugs) compared with spring and winter surveys. When winds were < 5 km/h, the arrival of adult T. infestans at the light traps was significantly associated with maximum temperature and relative humidity. This is the first field report of seasonal variations in the flight dispersal activity of T. infestans. PMID:17044877

  12. Differences in X-Chromosome Transcriptional Activity and Cholesterol Metabolism between Placentae from Swine Breeds from Asian and Western Origins

    PubMed Central

    Bischoff, Steve R.; Tsai, Shengdar Q.; Hardison, Nicholas E.; Motsinger-Reif, Alison A.; Freking, Bradley A.; Nonneman, Dan J.; Rohrer, Gary A.; Piedrahita, Jorge A.

    2013-01-01

    To gain insight into differences in placental physiology between two swine breeds noted for their dissimilar reproductive performance, that is, the Chinese Meishan and white composite (WC), we examined gene expression profiles of placental tissues collected at 25, 45, 65, 85, and 105 days of gestation by microarrays. Using a linear mixed model, a total of 1,595 differentially expressed genes were identified between the two pig breeds using a false-discovery rate q-value ≤0.05. Among these genes, we identified breed-specific isoforms of XIST, a long non-coding RNA responsible X-chromosome dosage compensation in females. Additionally, we explored the interaction of placental gene expression and chromosomal location by DIGMAP and identified three Sus scrofa X chromosomal bands (Xq13, Xq21, Xp11) that represent transcriptionally active clusters that differ between Meishan and WC during placental development. Also, pathway analysis identified fundamental breed differences in placental cholesterol trafficking and its synthesis. Direct measurement of cholesterol confirmed that the cholesterol content was significantly higher in the Meishan versus WC placentae. Taken together, this work identifies key metabolic pathways that differ in the placentae of two swine breeds noted for differences in reproductive prolificacy. PMID:23383161

  13. Detection of enterotoxigenic Clostridium perfringens in spices used in Mexico by dot blotting using a DNA probe.

    PubMed

    Rodríguez-Romo, L A; Heredia, N L; Labbé, R G; García-Alvarado, J S

    1998-02-01

    Several reports on the microbiology of spices and herbs indicate the presence of Clostridium perfringens, a spore-forming foodborne pathogen responsible for gastrointestinal disease. In the present study, a total of 380 samples of spices and herbs (cumin seed, black pepper, oregano, garlic powder, and bay leaves) widely used in Mexico were analyzed for the presence of C. perfringens, and the enterotoxigenicity of the isolates was determined by a dot-blot technique using an enterotoxin degoxigenin-labeled DNA probe. C. perfringens counts varied from <100 to 433 CFU/g in garlic powder, from <100 to 200 CFU/g in black pepper, from <100 to 433 CFU/g in cumin seed, from <100 to 340 CFU/g in oregano, and from < 100 to 450 CFU/g in bay leaves. The dot-blot technique detected the enterotoxin gene in 8 (4.25%) of 188 confirmed isolates of C. perfringens. dot-blot.

  14. HPTLC Plate Blotting for Liquid Microjunction Surface Sampling Probe Mass Spectrometric Analysis of Analytes Separated on a Wettable Phase Plate

    SciTech Connect

    Walworth, Matthew J; Stankovich, Joseph J; Van Berkel, Gary J; Schulz, Michael; Minarik, susanne

    2012-01-01

    A blotting method that transfers analytes separated on wettable HPTLC plates to a hydrophobic reversed-phase C8 HPLTC plate suitable for analysis with a liquid microjunction surface sampling probe electrospray ionization mass spectrometry system was described and demonstrated. The simple blotting procedure transfers the analyte from the wettable plate to the topmost surface of a rigidly backed, easy-to-mount hydrophobic substrate that already has been proven viable for analysis by this sampling probe/mass spectrometry system. The utility of the approach was demonstrated by the analysis of a four-component peptide mixture originally separated on a ProteoChrom HPTLC cellulose sheet and then blotted to the reversed phase HPTLC plate.

  15. Potential respiration estimated by electron transport system activity in deep-sea suprabenthic crustaceans off Balearic Islands (Western Mediterranean)

    NASA Astrophysics Data System (ADS)

    Herrera, A.; Gómez, M.; Packard, T. T.; Reglero, P.; Blanco, E.; Barberá-Cebrián, C.

    2014-10-01

    ETS is an acronym for the activity of the respiratory electron transport system; the ETS assay is a biochemical method for estimating the “potential” respiration (Φ). We apply this technique to suprabenthic species captured at three depths (250 m, 650 m and 850 m) in two different locations: Cabrera (Algerian subbasin) and Sóller (Balearic subbasin) during the IDEADOS survey during summer 2010. The aim of this study was to compare specific Φ between areas and between three depths to identify differences in the suprabenthos physiological state related to nutritional conditions. Specific Φ, expressed in unit of μl O2 h- 1 mg prot- 1 was not significantly different between species. Mean values were for the decapods: Plesionika heterocarpus, 8.4 ± 7.9; Gennadas elegans, 8.3 ± 2.9; and Sergestes arcticus 7.3 ± 4.6. Within the euphausiids specific Φ averaged 6.5 ± 4.2 for Thysanopoda aequalis and 9.8 ± 5.1 for Meganyctiphanes norvegica; while for the mysids it ranged from 7.7 ± 4.4 for Boreomysis arctica and 2.1 ± 0.6 for Eucopia unguiculata. The comparison of specific potential respiration (Φ), with the pooling of the data of all the species, showed differences between the two locations, being higher in Cabrera. However, no significant differences between the different depths of each locality were found. The slope of the log Φ-log biomass plot was 0.93 ± 0.09 for Cabrera and 0.64 ± 0.11 in Sóller. We interpret these differences as indicating that the suprabenthos in the Cabrera area, as compared to the Sóller area, has been well-nourished.

  16. Diffuse flow hydrothermal manganese mineralization along the active Mariana and southern Izu-Bonin arc system, western Pacific

    NASA Astrophysics Data System (ADS)

    Hein, James R.; Schulz, Marjorie S.; Dunham, Rachel E.; Stern, Robert J.; Bloomer, Sherman H.

    2008-08-01

    Abundant ferromanganese oxides were collected along 1200 km of the active Izu-Bonin-Mariana arc system. Chemical compositions and mineralogy show that samples were collected from two deposit types: Fe-Mn crusts of mixed hydrogenetic/hydrothermal origin and hydrothermal Mn oxide deposits; this paper addresses only the second type. Mn oxides cement volcaniclastic and biogenic sandstone and breccia layers (Mn sandstone) and form discrete dense stratabound layers along bedding planes and within beds (stratabound Mn). The Mn oxide was deposited within coarse-grained sediments from diffuse flow systems where precipitation occurred below the seafloor. Deposits were exposed at the seabed by faulting, mass wasting, and erosion. Scanning electron microscopy and microprobe analyses indicate the presence of both amorphous and crystalline 10 Å and 7 Å manganate minerals, the fundamental chemical difference being high water contents in the amorphous Mn oxides. Alternation of amorphous and crystalline laminae occurs in many samples, which likely resulted from initial rapid precipitation of amorphous Mn oxides from waxing pulses of hydrothermal fluids followed by precipitation of slow forming crystallites during waning stages. The chemical composition is characteristic of a hydrothermal origin including strong fractionation between Fe (mean 0.9 wt %) and Mn (mean 48 wt %) for the stratabound Mn, generally low trace metal contents, and very low rare earth element and platinum group element contents. However, Mo, Cd, Zn, Cu, Ni, and Co occur in high concentrations in some samples and may be good indicator elements for proximity to the heat source or to massive sulfide deposits. For the Mn sandstones, Fe (mean 8.4%) and Mn (12.4%) are not significantly fractionated because of high Fe contents in the volcaniclastic material. However, the proportion of hydrothermal Fe (nondetrital Fe) to total Fe is remarkably constant (49-58%) for all the sample groups, regardless of the degree of

  17. Diffuse flow hydrothermal manganese mineralization along the active Mariana and southern Izu-Bonin arc system, western Pacific

    USGS Publications Warehouse

    Hein, J.R.; Schulz, M.S.; Dunham, R.E.; Stern, R.J.; Bloomer, S.H.

    2008-01-01

    Abundant ferromanganese oxides were collected along 1200 km of the active Izu-Bonin-Mariana arc system. Chemical compositions and mineralogy show that samples were collected from two deposit types: Fe-Mn crusts of mixed hydrogenetic/hydrothermal origin and hydrothermal Mn oxide deposits; this paper addresses only the second type. Mn oxides cement volcaniclastic and biogenic sandstone and breccia layers (Mn sandstone) and form discrete dense stratabound layers along bedding planes and within beds (stratabound Mn). The Mn oxide was deposited within coarse-grained sediments from diffuse flow systems where precipitation occurred below the seafloor. Deposits were exposed at the seabed by faulting, mass wasting, and erosion. Scanning electron microscopy and microprobe analyses indicate the presence of both amorphous and crystalline 10 ?? and 7 ?? manganate minerals, the fundamental chemical difference being high water contents in the amorphous Mn oxides. Alternation of amorphous and crystalline laminae occurs in many samples, which likely resulted from initial rapid precipitation of amorphous Mn oxides from waxing pulses of hydrothermal fluids followed by precipitation of slow forming crystallites during waning stages. The chemical composition is characteristic of a hydrothermal origin including strong fractionation between Fe (mean 0.9 wt %) and Mn (mean 48 wt %) for the stratabound Mn, generally low trace metal contents, and very low rare earth element and platinum group element contents. However, Mo, Cd, Zn, Cu, Ni, and Co occur in high concentrations in some samples and may be good indicator elements for proximity to the heat source or to massive sulfide deposits. For the Mn sandstones, Fe (mean-8.4%) and Mn (12.4%) are not significantly fractionated because of high Fe contents in the volcaniclastic material. However, the proportion of hydrothermal Fe (nondetrital Fe) to total Fe is remarkably constant (49-58%) for all the sample groups, regardless of the degree of

  18. FIELD ACTIVITIES AND PRELIMINARY RESULTS FROM THE INVESTIGATION OF WESTERN AIRBORNE CONTAMINANTS IN TWO HIGH ELEVATION WATERSHEDS OF ROCKY MOUNTAIN NATIONAL PARK

    EPA Science Inventory

    The National Park Service initiated the Western Airborne Contaminants Assessment Project (WACAP) in 2002 to determine if airborne contaminants from long-range transport and/or regional sources are having an impact on remote western ecosystems, including AK. Rocky Mountain Nation...

  19. Spatial and Temporal Evolution of Eruptive Activity in a Youthful Extensional Setting: the Case of the Nyamulagira Volcanic Field, Western Branch of the East African Rift

    NASA Astrophysics Data System (ADS)

    Smets, B.; Kervyn, M.; d'Oreye, N.; Kervyn, F.

    2014-12-01

    Nyamulagira is the westernmost volcano of the Virunga volcanic province, in the western branch of the East African Rift. This shield volcano is one of the most active African volcanoes with one eruption every 1-4 year(s). Nyamulagira's eruptions usually occur along the flanks of the main edifice and in the lava plain, producing pyroclastic cone(s) and 10-20 km-long lava flows. Between 1913 and 1938, the activity was however restricted to the summit caldera, where lava fountains progressively gave birth to a lava lake, which disappeared in 1938 during the partial collapse of the summit caldera and the onset of a 2.5 years-long flank eruption. The location of flank eruptions and the orientation of the eruptive fissures are strongly influenced by the edifice loading, and by the NNW-SSE fracture network that crosses the main edifice and link it to the neighboring Nyiragongo volcano. But rift fault can also influence fissure orientations and cone alignments, especially for distal events. The flank eruptions typically have similar characteristics, lasting few days to few weeks, with an average of 20-30 days. Less frequently, flank eruptions can be larger and more complex, lasting several months and/or emitting much larger volumes of lava. By combining historical and recent observations, we suggest that magma overpressure at shallow depth is the main cause of flank events. Major eruptions seem to be related to a deeper source able to trigger large magma injections through deep structures, such as rift faults. Since April 2012, the activity of Nyamulagira is restricted to the summit caldera, with continuous and intense gas emissions and, since mid-2014, by lava fountains. This change in eruptive behavior, if it persists, may leads to the emergence of a new lava lake and may significantly decreases the frequency of flank events

  20. Use of sera from humans and dolphins with lacaziosis and sera from experimentally infected mice for Western Blot analyses of Lacazia loboi antigens.

    PubMed

    Mendoza, Leonel; Belone, Andréa F F; Vilela, Raquel; Rehtanz, Manuela; Bossart, Gregory D; Reif, John S; Fair, Patricia A; Durden, Wendy N; St Leger, Judy; Travassos, Luiz R; Rosa, Patricia S

    2008-01-01

    Antibodies in the sera of patients with lacaziosis recognized an approximately 193-kDa antigen and other Lacazia loboi antigens. Paracoccidioides brasiliensis gp43 antigen was detected by all evaluated sera, but they failed to detect a protein with the same molecular mass in L. loboi extracts. This study is the first to examine the humoral response to L. loboi antigens by using multiple host sera.

  1. A Western Blot-based Investigation of the Yeast Secretory Pathway Designed for an Intermediate-Level Undergraduate Cell Biology Laboratory

    ERIC Educational Resources Information Center

    Hood-DeGrenier, Jennifer K.

    2008-01-01

    The movement of newly synthesized proteins through the endomembrane system of eukaryotic cells, often referred to generally as the secretory pathway, is a topic covered in most intermediate-level undergraduate cell biology courses. An article previously published in this journal described a laboratory exercise in which yeast mutants defective in…

  2. Detection of PrPSc in Formalin Fixed Paraffin Embedded Tissue by Western Blot Differentiates Classical Scrapie, Nor98 Scrapie, and BSE

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Transmissible spongiform encephalopathies including bovine spongiform encephalopathy and scrapie are fatal neurodegenerative disorders associated with the presence of an infectious abnormal isoform of normal mammalian proteins called prions (PrP**Sc). Identification of PrP**Sc in the CNS is typicall...

  3. 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.

  4. Paleoflood activity and climate change over the last 2000 years recorded by high altitude alpine lake sediments in Western French Alps.

    NASA Astrophysics Data System (ADS)

    Fouinat, Laurent; Sabatier, Pierre; Develle, Anne-Lise; Giguet-Covex, Charline; Poulenard, Jérôme; Doyen, Elise; Crouzet, Christian; Malet, Emmanuel; Reyss, Jean-Louis; Arnaud, Fabien

    2015-04-01

    Extreme precipitation events can trigger floods that may have serious human and economic consequences. The flood represents extreme rainfall event, which in high altitude mountain regions are mostly triggered alternatively by local convective summer storms or, less frequently, by regional widespread rainfall event. The former's precipitation pattern comes from Mediterranean Sea fluxes, dominant in the south; instead of the latter coming from the Atlantic Ocean, dominant in the north of the French Alps. The aim of the study is then to explore which regime dominates in Western French Alps. Paleoflood chronicle is a way to understand past continental climate through the variability of both frequency and intensity. In this study we explore the paleoflood activity as recorded by sediments of the small alpine lake Muzelle (2200 m.a.s.l.) located in the western French Alps. Lake Muzelle catchment area is 5 km² -around 4 % being glacier covered- and is drained by one main stream. Lake Muzelle is 18.8 meters deep and is ice-covered during 7-8 months each year. Moreover, the watershed is being used for pastoral activity for several centuries. In this study, we use sedimentological analysis as well high resolution XRF core scanner geochemistry to identify turbidites interpreted as flood deposits. 256 turbidites were documented in the sediment sequence. Sr/Ti geochemical ratio is used to identify the coarsest grain size fraction of the flood deposit and the thickness of each deposit was measured. Dating was carried out using short-lived-radio-elements (210Pb, 137Cs, 241Am), historical events as well as nine 14C dates and paleomagnetic declination to constrain the age model over the last 2000 years. The study includes also palynological and sediment DNA analysis to understand past human activity on the watershed. As a result, the 31 years frequency shows a stable period from 0 to 1100 AD. Between 1100 and1200 AD the flood frequency presents a brutal increase with a relatively

  5. Methanogens at the top of the world: occurrence and potential activity of methanogens in newly deglaciated soils in high-altitude cold deserts in the Western Himalayas

    PubMed Central

    Aschenbach, Katrin; Conrad, Ralf; Řeháková, Klára; Doležal, Jiří; Janatková, Kateřina; Angel, Roey

    2013-01-01

    Methanogens typically occur in reduced anoxic environments. However, in recent studies it has been shown that many aerated upland soils, including desert soils also host active methanogens. Here we show that soil samples from high-altitude cold deserts in the western Himalayas (Ladakh, India) produce CH4 after incubation as slurry under anoxic conditions at rates comparable to those of hot desert soils. Samples of matured soil from three different vegetation belts (arid, steppe, and subnival) were compared with younger soils originating from frontal and lateral moraines of receding glaciers. While methanogenic rates were higher in the samples from matured soils, CH4 was also produced in the samples from the recently deglaciated moraines. In both young and matured soils, those covered by a biological soil crust (biocrust) were more active than their bare counterparts. Isotopic analysis showed that in both cases CH4 was initially produced from H2/CO2 but later mostly from acetate. Analysis of the archaeal community in the in situ soil samples revealed a clear dominance of sequences related to Thaumarchaeota, while the methanogenic community comprised only a minor fraction of the archaeal community. Similar to other aerated soils, the methanogenic community was comprised almost solely of the genera Methanosarcina and Methanocella, and possibly also Methanobacterium in some cases. Nevertheless, ~103 gdw−1 soil methanogens were already present in the young moraine soil together with cyanobacteria. Our results demonstrate that Methanosarcina and Methanocella not only tolerate atmospheric oxygen but are also able to survive in these harsh cold environments. Their occurrence in newly deglaciated soils shows that they are early colonizers of desert soils, similar to cyanobacteria, and may play a role in the development of desert biocrusts. PMID:24348469

  6. Methanogens at the top of the world: occurrence and potential activity of methanogens in newly deglaciated soils in high-altitude cold deserts in the Western Himalayas.

    PubMed

    Aschenbach, Katrin; Conrad, Ralf; Reháková, Klára; Doležal, Jiří; Janatková, Kateřina; Angel, Roey

    2013-01-01

    Methanogens typically occur in reduced anoxic environments. However, in recent studies it has been shown that many aerated upland soils, including desert soils also host active methanogens. Here we show that soil samples from high-altitude cold deserts in the western Himalayas (Ladakh, India) produce CH4 after incubation as slurry under anoxic conditions at rates comparable to those of hot desert soils. Samples of matured soil from three different vegetation belts (arid, steppe, and subnival) were compared with younger soils originating from frontal and lateral moraines of receding glaciers. While methanogenic rates were higher in the samples from matured soils, CH4 was also produced in the samples from the recently deglaciated moraines. In both young and matured soils, those covered by a biological soil crust (biocrust) were more active than their bare counterparts. Isotopic analysis showed that in both cases CH4 was initially produced from H2/CO2 but later mostly from acetate. Analysis of the archaeal community in the in situ soil samples revealed a clear dominance of sequences related to Thaumarchaeota, while the methanogenic community comprised only a minor fraction of the archaeal community. Similar to other aerated soils, the methanogenic community was comprised almost solely of the genera Methanosarcina and Methanocella, and possibly also Methanobacterium in some cases. Nevertheless, ~10(3) gdw(-1) soil methanogens were already present in the young moraine soil together with cyanobacteria. Our results demonstrate that Methanosarcina and Methanocella not only tolerate atmospheric oxygen but are also able to survive in these harsh cold environments. Their occurrence in newly deglaciated soils shows that they are early colonizers of desert soils, similar to cyanobacteria, and may play a role in the development of desert biocrusts.

  7. Fire activity as a function of fire–weather seasonal severity and antecedent climate across spatial scales in southern Europe and Pacific western USA

    USGS Publications Warehouse

    Urbieta, Itziar R.; Zavala, Gonzalo; Bedia, Joaquin; Gutierrez, Jose M.; San Miguel-Ayanz, Jesus; Camia, Andrea; Keeley, Jon E.; Moreno, Jose M.

    2015-01-01

    Climate has a strong influence on fire activity, varying across time and space. We analyzed the relationships between fire–weather conditions during the main fire season and antecedent water-balance conditions and fires in two Mediterranean-type regions with contrasted management histories: five southern countries of the European Union (EUMED)(all fires); the Pacific western coast of the USA (California and Oregon, PWUSA)(national forest fires). Total number of fires (≥1 ha), number of large fires (≥100 ha) and area burned were related to mean seasonal fire weather index (FWI), number of days over the 90th percentile of the FWI, and to the standardized precipitation-evapotranspiration index (SPEI) from the preceding 3 (spring) or 8 (autumn through spring) months. Calculations were made at three spatial aggregations in each area, and models related first-difference (year-to-year change) of fires and FWI/climate variables to minimize autocorrelation. An increase in mean seasonal FWI resulted in increases in the three fire variables across spatial scales in both regions. SPEI contributed little to explain fires, with few exceptions. Negative water-balance (dry) conditions from autumn through spring (SPEI8) were generally more important than positive conditions (moist) in spring (SPEI3), both of which contributed positively to fires. The R2 of the models generally improved with increasing area of aggregation. For total number of fires and area burned, the R2 of the models tended to decrease with increasing mean seasonal FWI. Thus, fires were more susceptible to change with climate variability in areas with less amenable conditions for fires (lower FWI) than in areas with higher mean FWI values. The relationships were similar in both regions, albeit weaker in PWUSA, probably due to the wider latitudinal gradient covered in PWUSA than in EUMED. The large variance explained by some of the models indicates that large-scale seasonal forecast could help anticipating

  8. Methanogens at the top of the world: occurrence and potential activity of methanogens in newly deglaciated soils in high-altitude cold deserts in the Western Himalayas.

    PubMed

    Aschenbach, Katrin; Conrad, Ralf; Reháková, Klára; Doležal, Jiří; Janatková, Kateřina; Angel, Roey

    2013-01-01

    Methanogens typically occur in reduced anoxic environments. However, in recent studies it has been shown that many aerated upland soils, including desert soils also host active methanogens. Here we show that soil samples from high-altitude cold deserts in the western Himalayas (Ladakh, India) produce CH4 after incubation as slurry under anoxic conditions at rates comparable to those of hot desert soils. Samples of matured soil from three different vegetation belts (arid, steppe, and subnival) were compared with younger soils originating from frontal and lateral moraines of receding glaciers. While methanogenic rates were higher in the samples from matured soils, CH4 was also produced in the samples from the recently deglaciated moraines. In both young and matured soils, those covered by a biological soil crust (biocrust) were more active than their bare counterparts. Isotopic analysis showed that in both cases CH4 was initially produced from H2/CO2 but later mostly from acetate. Analysis of the archaeal community in the in situ soil samples revealed a clear dominance of sequences related to Thaumarchaeota, while the methanogenic community comprised only a minor fraction of the archaeal community. Similar to other aerated soils, the methanogenic community was comprised almost solely of the genera Methanosarcina and Methanocella, and possibly also Methanobacterium in some cases. Nevertheless, ~10(3) gdw(-1) soil methanogens were already present in the young moraine soil together with cyanobacteria. Our results demonstrate that Methanosarcina and Methanocella not only tolerate atmospheric oxygen but are also able to survive in these harsh cold environments. Their occurrence in newly deglaciated soils shows that they are early colonizers of desert soils, similar to cyanobacteria, and may play a role in the development of desert biocrusts. PMID:24348469

  9. Seismicity and active tectonics in the Alboran Sea, Western Mediterranean: Constraints from an offshore-onshore seismological network and swath bathymetry data

    NASA Astrophysics Data System (ADS)

    Grevemeyer, Ingo; Gràcia, Eulàlia; Villaseñor, Antonio; Leuchters, Wiebke; Watts, Anthony B.

    2015-12-01

    Seismicity and tectonic structure of the Alboran Sea were derived from a large amphibious seismological network deployed in the offshore basins and onshore in Spain and Morocco, an area where the convergence between the African and Eurasian plates causes distributed deformation. Crustal structure derived from local earthquake data suggests that the Alboran Sea is underlain by thinned continental crust with a mean thickness of about 20 km. During the 5 months of offshore network operation, a total of 229 local earthquakes were located within the Alboran Sea and neighboring areas. Earthquakes were generally crustal events, and in the offshore domain, most of them occurred at crustal levels of 2 to 15 km depth. Earthquakes in the Alboran Sea are poorly related to large-scale tectonic features and form a 20 to 40 km wide NNE-SSW trending belt of seismicity between Adra (Spain) and Al Hoceima (Morocco), supporting the case for a major left-lateral shear zone across the Alboran Sea. Such a shear zone is in accord with high-resolution bathymetric data and seismic reflection imaging, indicating a number of small active fault zones, some of which offset the seafloor, rather than supporting a well-defined discrete plate boundary fault. Moreover, a number of large faults known to be active as evidenced from bathymetry, seismic reflection, and paleoseismic data such as the Yusuf and Carboneras faults were seismically inactive. Earthquakes below the Western Alboran Basin occurred at 70 to 110 km depth and hence reflected intermediate depth seismicity related to subducted lithosphere.

  10. Fire activity as a function of fire-weather seasonal severity and antecedent climate across spatial scales in southern Europe and Pacific western USA

    NASA Astrophysics Data System (ADS)

    Urbieta, Itziar R.; Zavala, Gonzalo; Bedia, Joaquín; Gutiérrez, José M.; San Miguel-Ayanz, Jesús; Camia, Andrea; Keeley, Jon E.; Moreno, José M.

    2015-11-01

    Climate has a strong influence on fire activity, varying across time and space. We analyzed the relationships between fire-weather conditions during the main fire season and antecedent water-balance conditions and fires in two Mediterranean-type regions with contrasted management histories: five southern countries of the European Union (EUMED)(all fires); the Pacific western coast of the USA (California and Oregon, PWUSA)(national forest fires). Total number of fires (≥1 ha), number of large fires (≥100 ha) and area burned were related to mean seasonal fire weather index (FWI), number of days over the 90th percentile of the FWI, and to the standardized precipitation-evapotranspiration index (SPEI) from the preceding 3 (spring) or 8 (autumn through spring) months. Calculations were made at three spatial aggregations in each area, and models related first-difference (year-to-year change) of fires and FWI/climate variables to minimize autocorrelation. An increase in mean seasonal FWI resulted in increases in the three fire variables across spatial scales in both regions. SPEI contributed little to explain fires, with few exceptions. Negative water-balance (dry) conditions from autumn through spring (SPEI8) were generally more important than positive conditions (moist) in spring (SPEI3), both of which contributed positively to fires. The R2 of the models generally improved with increasing area of aggregation. For total number of fires and area burned, the R2 of the models tended to decrease with increasing mean seasonal FWI. Thus, fires were more susceptible to change with climate variability in areas with less amenable conditions for fires (lower FWI) than in areas with higher mean FWI values. The relationships were similar in both regions, albeit weaker in PWUSA, probably due to the wider latitudinal gradient covered in PWUSA than in EUMED. The large variance explained by some of the models indicates that large-scale seasonal forecast could help anticipating fire

  11. A downscaling technique to simulate changes in western North Pacific tropical cyclone activity between two types of El Niño events

    NASA Astrophysics Data System (ADS)

    Zhao, Haikun

    2016-02-01

    Changes in tropical cyclone (TC) activity over the western North Pacific (WNP) basin between the Modoki and canonical El Niño years have been simulated and examined in this study based on a downscaling technique. The downscaling technique is used for generating synthetic TCs based on TC formation, TC track, and TC intensity models. Results suggest that the downscaling technique can well simulate the spatial distribution of TC activity during the two types of El Niño years and their differences. It is found that the observed changes in TC tracks during the two types of El Niño years are mainly due to the combined effects of changes in TC formation locations and large-scale steering flows. Further examinations have shown that changes in large-scale steering flows play a more important role than changes in TC formation locations. These results are in accordance with the cyclonic circulation anomaly found during the Modoki El Niño years compared to that during the canonical El Niño years. Numerical simulations further suggest that changes in TC tracks between the two types of El Niño years appear to be the most important factor affecting the TC intensity change. Compared to that during the Modoki El Niño years, TC formation is enhanced in the south quadrant of the WNP basin and more TCs take a northwestward track during the canonical El Niño years, leading to a longer TC lifespan and greater TC intensity.

  12. Evaluation of cytochrome P450 2C9 activity in normal, healthy, adult Western Indian population by both phenotyping and genotyping

    PubMed Central

    Swar, Balkrishna D.; Bendkhale, Shital R.; Rupawala, Abbas; Sridharan, Kannan; Gogtay, Nithya J.; Thatte, Urmila M.; Kshirsagar, Nilima A.

    2016-01-01

    Objectives: Cytochrome P450 2C9 (CYP2C9) is a member of cytochrome P450 (CYP) family that accounts for nearly 18% of the total CYP protein content in the human liver microsomes and catalyzes almost 15–20% of the drugs. Considering the paucity of data on the polymorphisms of CYP2C9 in Western Indian population, the present study was conducted to evaluate the prevalence of CYP2C9 polymorphisms (*1, *2 and *3) and correlate it with the activity using flurbiprofen (FLB) as a probe drug. Materials and Methods: A 100 mg FLB capsule was administered to 298 healthy adult participants. Venous blood samples were analyzed at 2 h postdose for the estimation of FLB and 4-hydroxy FLB. Metabolic ratio (MR) was calculated to determine the extent of poor metabolizer (PM) and rapid metabolizer status using probit plot. Genotyping of CYP2C9 polymorphism was performed using polymerase chain reaction-restriction fragment length polymorphism technique. Results: Of the total 298 participants, phenotype was assessable in 288 and genotype was performed in 289 participants. The median (range) MR of the study population was 6.6 (1.65–66.05). Five participants were found to be PMs by phenotype. Of the total 289 participants, 209 (72.3%) (66.7, 77.2) had CYP2C9*1/*1, 25 (8.7%) (5.8, 12.7) with CYP2C9*1/*2, 55 (19%) (14.8, 24.1) had CYP2C9*1/*3, 3 (1%) (0.3, 3.3) had CYP2C9*2/*3 genotype. A significant association between phenotype and genotype was observed. Conclusion: To conclude, the present study found significant association of CYP2C9 activity by both phenotype and genotype and these findings have to be corroborated in different kinds of patients. PMID:27298492

  13. Development of a Bayesian approach to assess the probability of concealed active faults using Helium isotope ratios: An example from the western Tottori area, SW Japan

    NASA Astrophysics Data System (ADS)

    Martin, A. J.; Umeda, K.; Ishimaru, T.

    2012-12-01

    In Japan, numerous research projects have been carried out to assess the long-term stability of the geological environment including in particular, the spatio-temporal distribution of volcanism and active faulting in the context siting of a geological repository for the long-term (0.1 - 1 Ma) disposal of high-level radioactive waste. Of particular concern is the existence of active faulting that shows no expression at the surface and thus not necessarily recorded in currently published active fault maps. Recent research carried out in the western Tottori area, southwest Japan, has implied the potential of using He-3/He-4 ratios as a means of providing indirect evidence of the existence of source fault(s) that caused the 6 Oct 2000 Tottori earthquake but which had no apparent surface indication [e.g. Umeda et al. 2011]. This is based on the theory that mantle helium can reach the shallow crustal depths due to faults in the crust acting as pathways [e.g. Kennedy et al. 1997]. Here we introduce a new technique based on Bayesian inference in an effort to quantify this phenomenon and present our preliminary findings. In the Bayesian paradigm, we make a priori assumptions based on the tectonic setting of the study area as a starting point. One a priori assumption could be that 'unknown' faults do not exist far from 'known' faults. Depending on the degree of conservation required in the first step, 2-D a priori probability distributions are calculated using kernel functions with varying values of standard deviation. The recently acquired helium isotope ratios are then combined to the a priori probability distributions using Bayes' rule to produce modified a posteriori probabilities that is expected to better capture the hidden faults. The potential of the methodology to incorporate other information such as GPS data will also be presented and discussed. References Kennedy, B. M., Y. K. Kharaka, W. C. Evans, A. Ellwood, D. J. DePaolo, J. Thordsen, G. Ambats, and R. H

  14. Characterizing the Mechanism of Action of Double-Stranded RNA Activity against Western Corn Rootworm (Diabrotica virgifera virgifera LeConte)

    PubMed Central

    Bolognesi, Renata; Ramaseshadri, Parthasarathy; Anderson, Jerry; Bachman, Pamela; Clinton, William; Flannagan, Ronald; Ilagan, Oliver; Lawrence, Christina; Levine, Steven; Moar, William; Mueller, Geoffrey; Tan, Jianguo; Uffman, Joshua; Wiggins, Elizabeth; Heck, Gregory; Segers, Gerrit

    2012-01-01

    RNA interference (RNAi) has previously been shown to be effective in western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte) larvae via oral delivery of synthetic double-stranded RNA (dsRNA) in an artificial diet bioassay, as well as by ingestion of transgenic corn plant tissues engineered to express dsRNA. Although the RNAi machinery components appear to be conserved in Coleopteran insects, the key steps in this process have not been reported for WCR. Here we characterized the sequence of events that result in mortality after ingestion of a dsRNA designed against WCR larvae. We selected the Snf7 ortholog (DvSnf7) as the target mRNA, which encodes an essential protein involved in intracellular trafficking. Our results showed that dsRNAs greater than or equal to approximately 60 base-pairs (bp) are required for biological activity in artificial diet bioassays. Additionally, 240 bp dsRNAs containing a single 21 bp match to the target sequence were also efficacious, whereas 21 bp short interfering (si) RNAs matching the target sequence were not. This result was further investigated in WCR midgut tissues: uptake of 240 bp dsRNA was evident in WCR midgut cells while a 21 bp siRNA was not, supporting the size-activity relationship established in diet bioassays. DvSnf7 suppression was observed in a time-dependent manner with suppression at the mRNA level preceding suppression at the protein level when a 240 bp dsRNA was fed to WCR larvae. DvSnf7 suppression was shown to spread to tissues beyond the midgut within 24 h after dsRNA ingestion. These events (dsRNA uptake, target mRNA and protein suppression, systemic spreading, growth inhibition and eventual mortality) comprise the overall mechanism of action by which DvSnf7 dsRNA affects WCR via oral delivery and provides insights as to how targeted dsRNAs in general are active against insects. PMID:23071820

  15. Lightning in Western Patagonia

    NASA Astrophysics Data System (ADS)

    Garreaud, René D.; Gabriela Nicora, M.; Bürgesser, Rodrigo E.; Ávila, Eldo E.

    2014-04-01

    On the basis of 8 years (2005-2012) of stroke data from the World Wide Lightning Location Network we describe the spatial distribution and temporal variability of lightning activity over Western Patagonia. This region extends from ~40°S to 55°S along the west coast of South America, is limited to the east by the austral Andes, and features a hyper-humid, maritime climate. Stroke density exhibits a sharp maximum along the coast of southern Chile. Although precipitation there is largely produced by cold nimbostratus, days with more than one stroke occur up to a third of the time somewhere along the coastal strip. Disperse strokes are also observed off southern Chile. In contrast, strokes are virtually nonexistent over the austral Andes—where precipitation is maximum—and farther east over the dry lowlands of Argentina. Atmospheric reanalysis and satellite imagery are used to characterize the synoptic environment of lightning-producing storms, exemplified by a case study and generalized by a compositing analysis. Lightning activity tends to occur when Western Patagonia is immersed in a pool of cold air behind a front that has reached the coast at ~40°S. Under these circumstances, midlevel cooling occurs before and is more prominent than near-surface cooling, leading to a weakly unstable postfrontal condition. Forced uplift of the strong westerlies impinging on the coastal mountains can trigger convection and produces significant lightning activity in this zone. Farther offshore, large-scale ascent near the cyclone's center may lift near-surface air parcels, fostering shallow convection and dispersing lightning activity.

  16. Polymorphisms in Pfmdr1, Pfcrt, and Pfnhe1 genes are associated with reduced in vitro activities of quinine in Plasmodium falciparum isolates from western Kenya.

    PubMed

    Cheruiyot, Jelagat; Ingasia, Luicer A; Omondi, Angela A; Juma, Dennis W; Opot, Benjamin H; Ndegwa, Joseph M; Mativo, Joan; Cheruiyot, Agnes C; Yeda, Redemptah; Okudo, Charles; Muiruri, Peninah; Bidii, Ngalah S; Chebon, Lorna J; Angienda, Paul O; Eyase, Fredrick L; Johnson, Jacob D; Bulimo, Wallace D; Andagalu, Ben; Akala, Hoseah M; Kamau, Edwin

    2014-07-01

    In combination with antibiotics, quinine is recommended as the second-line treatment for uncomplicated malaria, an alternative first-line treatment for severe malaria, and for treatment of malaria in the first trimester of pregnancy. Quinine has been shown to have frequent clinical failures, and yet the mechanisms of action and resistance have not been fully elucidated. However, resistance is linked to polymorphisms in multiple genes, including multidrug resistance 1 (Pfmdr1), the chloroquine resistance transporter (Pfcrt), and the sodium/hydrogen exchanger gene (Pfnhe1). Here, we investigated the association between in vitro quinine susceptibility and genetic polymorphisms in Pfmdr1codons 86 and 184, Pfcrt codon 76, and Pfnhe1 ms4760 in 88 field isolates from western Kenya. In vitro activity was assessed based on the drug concentration that inhibited 50% of parasite growth (the IC50), and parasite genetic polymorphisms were determined from DNA sequencing. Data revealed there were significant associations between polymorphism in Pfmdr1-86Y, Pfmdr1-184F, or Pfcrt-76T and quinine susceptibility (P < 0.0001 for all three associations). Eighty-two percent of parasites resistant to quinine carried mutant alleles at these codons (Pfmdr1-86Y, Pfmdr1-184F, and Pfcrt-76T), whereas 74% of parasites susceptible to quinine carried the wild-type allele (Pfmdr1-N86, Pfmdr1-Y184, and Pfcrt-K76, respectively). In addition, quinine IC50 values for parasites with Pfnhe1 ms4760 3 DNNND repeats were significantly higher than for those with 1 or 2 repeats (P = 0.033 and P = 0.0043, respectively). Clinical efficacy studies are now required to confirm the validity of these markers and the importance of parasite genetic background.

  17. Chemical composition and antimicrobial activity of the essential oil of Ocimum basilicum L. (sweet basil) from Western Ghats of North West Karnataka, India

    PubMed Central

    Joshi, Rajesh K.

    2014-01-01

    Context: Ocimum basilicum L. (Lamiaceae) commonly known as sweet basil, has been used as a traditional medicinal plant for the treatment of headaches, coughs, diarrhea, constipation, warts, worms, and kidney malfunctions. Materials and Methods: The essential oil of the flowering aerial parts of O. basilicum growing in the Western Ghats region of North West Karnataka, India, was obtained by hydro-distillation and analyzed by gas chromatography equipped with flame ionization detector and gas chromatography coupled to mass spectrometry (GC–MS). The oil was tested against six Gram-positive, eight Gram-negative bacteria, and three fungi by the tube-dilution method at a concentration range of 5.00-0.009 mg/mL. Results: Twenty-five constituents were identified in the essential oil of O. basilicum. The major constituents were identified as methyl eugenol (39.3%) and methyl chavicol (38.3%), accounting for 98.6% of the total oil. The oil was found to be active against Gram-positive, Gram-negative bacteria, and fungi with minimal bactericidal concentration values in the range of 0.143 ± 0.031 to 0.572 ± 0.127 mg/mL, 0.781 ± 0.382 to 1.875 ± 0.684 mg/mL, and 0.312 ± 0.171 to 0.442 ± 0.207 mg/mL, respectively. Conclusion: The essential oil of O. basilicum of this region contains methyl eugenol/methyl chavicol chemotype and has bactericidal properties. PMID:25538349

  18. New zircon data supporting models of short-lived igneous activity at 1.89 Ga in the western Skellefte District, central Fennoscandian Shield

    NASA Astrophysics Data System (ADS)

    Skyttä, P.; Hermansson, T.; Andersson, J.; Weihed, P.

    2011-04-01

    New U-Th-Pb zircon data (SIMS) from three intrusive phases of the Palaeoproterozoic Viterliden intrusion in the western Skellefte District, central Fennoscandian Shield, dates igneous emplacement in a narrow time interval at about 1.89 Ga. A locally occurring quartz-plagioclase porphyritic tonalite, here dated at 1889 ± 3 Ma, is, based on the new age data and field evidence, considered the youngest of the intrusive units. This supports an existing interpretation of its fault-controlled emplacement after intrusion of the dominating hornblende-tonalite units, in this study dated at 1892 ± 3 Ma. The Viterliden magmatism was synchronous with the oldest units of the Jörn type early-orogenic intrusions in the eastern part of the district (1.89-1.88 Ga; cf. Gonzàles Roldán, 2010). A U-Pb zircon age for a felsic metavolcanic rock from the hanging-wall to the Kristineberg VMS deposit, immediately south of the Viterliden intrusion, is in this study constrained in the 1.89-1.88 Ga time interval. It provides a minimum age for the Kristineberg ore deposit and suggests contemporaneous igneous/volcanic activity throughout the Skellefte District. Furthermore, it supports the view that the Skellefte Group defines a laterally continuous belt throughout this "ore district". Tentative correlation of the 1889 ± 3 Ma quartz-plagioclase porphyritic tonalite with the Kristineberg "mine porphyry", which cuts the altered ore-hosting metavolcanic rocks, further constrain the minimum age for ore deposition at 1889 ± 3 Ma. Based on the new age determinations, the Viterliden intrusion may equally well have intruded into, or locally acted as a basement for the ore-hosting Skellefte Group volcanic rocks.

  19. New zircon data supporting models of short-lived igneous activity at 1.89 Ga in the western Skellefte District, central Fennoscandian Shield

    NASA Astrophysics Data System (ADS)

    Skyttä, P.; Hermansson, T.; Andersson, J.; Whitehouse, M.; Weihed, P.

    2011-10-01

    New U-Th-Pb zircon data (SIMS) from three intrusive phases of the Palaeoproterozoic Viterliden intrusion in the western Skellefte District, central Fennoscandian Shield, dates igneous emplacement in a narrow time interval at about 1.89 Ga. A locally occurring quartz-plagioclase porphyritic tonalite, here dated at 1889 ± 3 Ma, is considered the youngest of the intrusive units, based on the new age data and field evidence. This supports an existing interpretation of its fault-controlled emplacement after intrusion of the dominating hornblende-tonalite units, in this study dated at 1892 ± 3 Ma. The Viterliden magmatism was synchronous with the oldest units of the Jörn type early-orogenic intrusions in the eastern part of the district (1.89-1.88 Ga; cf. Gonzàles Roldán, 2010). A U-Pb zircon age for a felsic metavolcanic rock from the hanging-wall to the Kristineberg VMS deposit, immediately south of the Viterliden intrusion, is constrained at 1883 ± 6 Ma in this study. It provides a minimum age for the Kristineberg ore deposit and suggests contemporaneous igneous/volcanic activity throughout the Skellefte District. Furthermore, it supports the view that the Skellefte Group defines a laterally continuous belt throughout this "ore district". Tentative correlation of the 1889 ± 3 Ma quartz-plagioclase porphyritic tonalite with the Kristineberg "mine porphyry" suggests that these units are coeval at about 1.89 Ga. Based on the new age determinations, the Viterliden intrusion may equally well have intruded into or locally acted as a basement for the ore-hosting Skellefte Group volcanic rocks.

  20. Snapshot blotting: transfer of nucleic acids and nucleoprotein complexes from electrophoresis gels to grids for electron microscopy.

    PubMed Central

    Jett, S D; Bear, D G

    1994-01-01

    We present a technique, "snapshot blotting," for the electrophoretic transfer of nucleic acids and nucleoprotein complexes in gel electrophoresis bands onto highly stable carbon film-coated grids for imaging by electron microscopy. The method permits structural analysis of macromolecular species that have been resolved by a gel mobility-shift assay. To demonstrate the efficiency and integrity of the transfer process for a multiprotein-DNA assembly, we have imaged various species of a prokaryotic transcription complex, using the cleavage-defective EcoRI(Q111) protein as an orientation marker and as a blockade of transcription elongation. Snapshot blotting should be of great utility in the structural characterization of nucleic acids and protein-nucleic acid interactions. Images PMID:8041711

  1. Differentiation of Enterococcus faecium from Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus strains by PCR and dot-blot hybridisation.

    PubMed

    Langa, S; Fernández, A; Martín, R; Reviriego, C; Marín, M L; Fernández, L; Rodríguez, J M

    2003-12-01

    Variations in length and sequence of the 16S/23S spacer region of Enterococcus faecium provided the basis for development of simple PCR and dot-blot hybridisation assays that enabled the differentiation of potentially probiotic Enterococcus faecium strains from Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. Such assays may be useful for differentiation of yoghurt starter cultures and enterococcal strains when they are simultaneously present in probiotic food products.

  2. Molecular Phenotyping Small (Asian) versus Large (Western) Plaque Psoriasis Shows Common Activation of IL-17 Pathway Genes, but Different Regulatory Gene Sets

    PubMed Central

    Kim, Jaehwan; Oh, Chil-Hwan; Jeon, Jiehyun; Baek, Yoosang; Ahn, Jaewoo; Kim, Dong Joo; Lee, Hyun-Soo; da Rosa, Joel Correa; Suárez-Fariñas, Mayte; Lowes, Michelle A.; Krueger, James G.

    2015-01-01

    Psoriasis is present in all racial groups, but in varying frequencies and severity. Considering that small plaque psoriasis is specific to the Asian population and severe psoriasis is more predominant in the Western population, we defined Asian small and intermediate plaque psoriasis as psoriasis subtypes, and compared their molecular signatures with classic subtype of Western large plaque psoriasis. Two different characteristics of psoriatic spreading—vertical growth and radial expansion—were contrasted between subtypes, and genomic data were correlated to histologic and clinical measurements. Compared to Western large plaque psoriasis, Asian small plaque psoriasis revealed limited psoriasis spreading, but IL-17A and IL-17-regulated pro-inflammatory cytokines were highly expressed. Paradoxically, IL-17A and IL-17-regulated pro-inflammatory cytokines were lower in Western large plaque psoriasis, while T cells and dendritic cells in total psoriatic skin area were exponentially increased. Negative immune regulators, such as CD69 and FAS, were decreased in both Western large plaque psoriasis and psoriasis with accompanying arthritis or obesity, and their expression was correlated with psoriasis severity index. Based on the disease subtype comparisons, we propose that dysregulation of T cell expansion enabled by downregulation of immune negative regulators is the main mechanism for development of large plaque psoriasis subtypes. PMID:26763436

  3. Molecular Phenotyping Small (Asian) versus Large (Western) Plaque Psoriasis Shows Common Activation of IL-17 Pathway Genes but Different Regulatory Gene Sets.

    PubMed

    Kim, Jaehwan; Oh, Chil-Hwan; Jeon, Jiehyun; Baek, Yoosang; Ahn, Jaewoo; Kim, Dong Joo; Lee, Hyun-Soo; Correa da Rosa, Joel; Suárez-Fariñas, Mayte; Lowes, Michelle A; Krueger, James G

    2016-01-01

    Psoriasis is present in all racial groups, but in varying frequencies and severity. Considering that small plaque psoriasis is specific to the Asian population and severe psoriasis is more predominant in the Western population, we defined Asian small and intermediate plaque psoriasis as psoriasis subtypes and compared their molecular signatures with the classic subtype of Western large plaque psoriasis. Two different characteristics of psoriatic spreading-vertical growth and radial expansion-were contrasted between subtypes, and genomic data were correlated to histologic and clinical measurements. Compared with Western large plaque psoriasis, Asian small plaque psoriasis revealed limited psoriasis spreading, but IL-17A and IL-17-regulated proinflammatory cytokines were highly expressed. Paradoxically, IL-17A and IL-17-regulated proinflammatory cytokines were lower in Western large plaque psoriasis, whereas T cells and dendritic cells in total psoriatic skin area were exponentially increased. Negative immune regulators, such as CD69 and FAS, were decreased in both Western large plaque psoriasis and psoriasis with accompanying arthritis or obesity, and their expression was correlated with psoriasis severity index. Based on the disease subtype comparisons, we propose that dysregulation of T-cell expansion enabled by downregulation of immune negative regulators is the main mechanism for development of large plaque psoriasis subtypes. PMID:26763436

  4. Molecular Phenotyping Small (Asian) versus Large (Western) Plaque Psoriasis Shows Common Activation of IL-17 Pathway Genes but Different Regulatory Gene Sets.

    PubMed

    Kim, Jaehwan; Oh, Chil-Hwan; Jeon, Jiehyun; Baek, Yoosang; Ahn, Jaewoo; Kim, Dong Joo; Lee, Hyun-Soo; Correa da Rosa, Joel; Suárez-Fariñas, Mayte; Lowes, Michelle A; Krueger, James G

    2016-01-01

    Psoriasis is present in all racial groups, but in varying frequencies and severity. Considering that small plaque psoriasis is specific to the Asian population and severe psoriasis is more predominant in the Western population, we defined Asian small and intermediate plaque psoriasis as psoriasis subtypes and compared their molecular signatures with the classic subtype of Western large plaque psoriasis. Two different characteristics of psoriatic spreading-vertical growth and radial expansion-were contrasted between subtypes, and genomic data were correlated to histologic and clinical measurements. Compared with Western large plaque psoriasis, Asian small plaque psoriasis revealed limited psoriasis spreading, but IL-17A and IL-17-regulated proinflammatory cytokines were highly expressed. Paradoxically, IL-17A and IL-17-regulated proinflammatory cytokines were lower in Western large plaque psoriasis, whereas T cells and dendritic cells in total psoriatic skin area were exponentially increased. Negative immune regulators, such as CD69 and FAS, were decreased in both Western large plaque psoriasis and psoriasis with accompanying arthritis or obesity, and their expression was correlated with psoriasis severity index. Based on the disease subtype comparisons, we propose that dysregulation of T-cell expansion enabled by downregulation of immune negative regulators is the main mechanism for development of large plaque psoriasis subtypes.

  5. Resistance evolution to the first generation of genetically modified Diabrotica-active Bt-maize events by western corn rootworm: management and monitoring considerations.

    PubMed

    Devos, Yann; Meihls, Lisa N; Kiss, József; Hibbard, Bruce E

    2013-04-01

    Western corn rootworm (Diabrotica virgifera virgifera; WCR) is a major coleopteran maize pest in North America and the EU, and has traditionally been managed through crop rotation and broad-spectrum soil insecticides. Genetically modified Bt-maize offers an additional management tool for WCR and has been valuable in reducing insecticide use and increasing farm income. A concern is that the widespread, repeated, and exclusive deployment of the same Bt-maize transformation event will result in the rapid evolution of resistance in WCR. This publication explores the potential of WCR to evolve resistance to plant-produced Bt-toxins from the first generation of Diabrotica-active Bt-maize events (MON 863 and MON 88017, DAS-59122-7 and MIR604), and whether currently implemented risk management strategies to delay and monitor resistance evolution are appropriate. In twelve of the twelve artificial selection experiments reported, resistant WCR populations were yielded rapidly. Field-selected resistance of WCR to Cry3Bb1 is documented in some US maize growing areas, where an increasing number of cases of unexpected damage of WCR larvae to Bt-maize MON 88017 has been reported. Currently implemented insect resistance management measures for Bt-crops usually rely on the high dose/refuge (HDR) strategy. Evidence (including laboratory, greenhouse and field data) indicates that several conditions contributing to the success of the HDR strategy may not be met for the first generation of Bt-maize events and WCR: (1) the Bt-toxins are expressed heterogeneously at a low-to-moderate dose in roots; (2) resistance alleles may be present at a higher frequency than initially assumed; (3) WCR may mate in a non-random manner; (4) resistance traits could have non-recessive inheritance; and (5) fitness costs may not necessarily be associated with resistance evolution. However, caution must be exercised when extrapolating laboratory and greenhouse results to field conditions. Model predictions

  6. Resistance evolution to the first generation of genetically modified Diabrotica-active Bt-maize events by western corn rootworm: management and monitoring considerations.

    PubMed

    Devos, Yann; Meihls, Lisa N; Kiss, József; Hibbard, Bruce E

    2013-04-01

    Western corn rootworm (Diabrotica virgifera virgifera; WCR) is a major coleopteran maize pest in North America and the EU, and has traditionally been managed through crop rotation and broad-spectrum soil insecticides. Genetically modified Bt-maize offers an additional management tool for WCR and has been valuable in reducing insecticide use and increasing farm income. A concern is that the widespread, repeated, and exclusive deployment of the same Bt-maize transformation event will result in the rapid evolution of resistance in WCR. This publication explores the potential of WCR to evolve resistance to plant-produced Bt-toxins from the first generation of Diabrotica-active Bt-maize events (MON 863 and MON 88017, DAS-59122-7 and MIR604), and whether currently implemented risk management strategies to delay and monitor resistance evolution are appropriate. In twelve of the twelve artificial selection experiments reported, resistant WCR populations were yielded rapidly. Field-selected resistance of WCR to Cry3Bb1 is documented in some US maize growing areas, where an increasing number of cases of unexpected damage of WCR larvae to Bt-maize MON 88017 has been reported. Currently implemented insect resistance management measures for Bt-crops usually rely on the high dose/refuge (HDR) strategy. Evidence (including laboratory, greenhouse and field data) indicates that several conditions contributing to the success of the HDR strategy may not be met for the first generation of Bt-maize events and WCR: (1) the Bt-toxins are expressed heterogeneously at a low-to-moderate dose in roots; (2) resistance alleles may be present at a higher frequency than initially assumed; (3) WCR may mate in a non-random manner; (4) resistance traits could have non-recessive inheritance; and (5) fitness costs may not necessarily be associated with resistance evolution. However, caution must be exercised when extrapolating laboratory and greenhouse results to field conditions. Model predictions

  7. Geology of the Crust and Mantle, Western United States: Geophysical data reveal a thin crust and anomalous upper mantle characteristic of active regions.

    PubMed

    Thompson, G A; Talwani, M

    1964-12-18

    compressional and transverse waves in the upper mantle may be related to this problem. Whatever its origin and composition, an anomalous upper mantle characterizes many regions of present or recent tectonic activity, such as Japan and the Mid-Atlantic Ridge (39). The anomalous mantle of western North America might form a continuous belt to the south, with anomalous mantle beneath the crest of the East Pacific Rise (40). The anomalous upper mantle may thus be an essential part of the heat engine driving the tectonic activity of these regions. The Basin and Range region was broken into blocks and laterally extended during the Cenozoic uplift, so that some blocks lagged behind, or sank. Some of the intricate disruption of the upper crust may be related to shallow Cenozoic volcanism. The relatively large and rigid Sierra Nevada block may have been tilted westward during Basin-Range deformation because of the high density of greenstones on the west side and the lower density of granitic rocks to the east. Man's environment, in the longer view of geologic time, is strongly influenced by mountain-building processes originating in the earth's crust and mantle. In the scale of a few lifetimes, climate, sea level, and the shape of the land are appreciably altered. How this comes about, and whether man can hope to influence the processes, are challenging, unsolved problems. But enough has now been learned about the crust and mantle to suggest precisely what questions must be answered and what critical experiments performed. Note added in proof: Osborne (42) has directed our attention to the possibility that the granitic rocks and also the andesites and dacites were formed by fractional crystallization of basaltic magma under conditions of high oxygen pressure. This possibility in no way conflicts with the geophysical data. In fact, such direct additions to the silicic upper crust from the mantle or lower crust would simplify the perplexing problem of how the crust is replenished in areas

  8. Geology of the Crust and Mantle, Western United States: Geophysical data reveal a thin crust and anomalous upper mantle characteristic of active regions.

    PubMed

    Thompson, G A; Talwani, M

    1964-12-18

    compressional and transverse waves in the upper mantle may be related to this problem. Whatever its origin and composition, an anomalous upper mantle characterizes many regions of present or recent tectonic activity, such as Japan and the Mid-Atlantic Ridge (39). The anomalous mantle of western North America might form a continuous belt to the south, with anomalous mantle beneath the crest of the East Pacific Rise (40). The anomalous upper mantle may thus be an essential part of the heat engine driving the tectonic activity of these regions. The Basin and Range region was broken into blocks and laterally extended during the Cenozoic uplift, so that some blocks lagged behind, or sank. Some of the intricate disruption of the upper crust may be related to shallow Cenozoic volcanism. The relatively large and rigid Sierra Nevada block may have been tilted westward during Basin-Range deformation because of the high density of greenstones on the west side and the lower density of granitic rocks to the east. Man's environment, in the longer view of geologic time, is strongly influenced by mountain-building processes originating in the earth's crust and mantle. In the scale of a few lifetimes, climate, sea level, and the shape of the land are appreciably altered. How this comes about, and whether man can hope to influence the processes, are challenging, unsolved problems. But enough has now been learned about the crust and mantle to suggest precisely what questions must be answered and what critical experiments performed. Note added in proof: Osborne (42) has directed our attention to the possibility that the granitic rocks and also the andesites and dacites were formed by fractional crystallization of basaltic magma under conditions of high oxygen pressure. This possibility in no way conflicts with the geophysical data. In fact, such direct additions to the silicic upper crust from the mantle or lower crust would simplify the perplexing problem of how the crust is replenished in areas

  9. Utility of slot-blot-ELISA as a new, fast, and sensitive immunoassay for detection of carcinoembryonic antigen in the urine samples of patients with various gastrointestinal malignancies.

    PubMed

    El-Masry, Samir; El-Sayed, Ibrahim H; Lotfy, Mahmoud; Mahmoud, Lamiaa; El-Naggar, Mohamed

    2007-01-01

    Carcinoembryonic antigen (CEA) is the most widely used clinical tumor marker. CEA immunoassay has found acceptance as a diagnostic adjunct in clinical diagnosis of gastrointestinal tumors (GIT). Several immunoassays have been established for detection of CEA in plasma, serum, tissue, feces, and urine of cancer patients using polyclonal or monoclonal antibodies raised against CEA. Some of these assays display both high sensitivity and specificity for the detection of CEA. However, these assays require special and highly expensive equipment and the procedures require long periods for their completion. In the present study, we established a Slot-Blot Enzyme Linked Immunosorbent Assay (SB-ELISA), based on anti-CEA monoclonal antibody (CEA-mAb), as a new, simple, fast, cheap, and non-invasive immunodiagnostic technique for detection of CEA in the urine of GIT patients. Urine and serum samples were collected from 248 GIT patients (58 with pancreatic cancer, 20 with hepatoma, 23 with ampullary carcinoma, 15 with hilar cholangiocarcinoma, 28 with gastric cancer, 14 with esophageal cancer, and 90 with colorectal cancer). Moreover, urine and serum samples were collected from 50 healthy individuals to serve as negative controls. The traditional ELISA technique was used for determination of CEA in the sera of GIT patients using anti-CEA monoclonal antibody. A comparison between the results of both techniques (ELISA and SB-ELISA) was carried out. The traditional ELISA detected CEA in the sera of 154 out of 248 GIT patients with a sensitivity of 59.8%, 51.7% positive predictive value (PPV) and 75.37% negative predictive value (NPV). In addition, it identified 15 false positive cases out of 50 healthy individuals with a specificity of 70%. The urinary CEA was identified by a Western blotting technique and CEA-mAb at a molecular mass of 180 Kda. The developed SB-ELISA showed higher sensitivity, specificity, PPV, and NPV (70.1%, 78%, 62.4%, and 82.13%, respectively) for detection

  10. Coriolus versicolor mushroom polysaccharides exert immunoregulatory effects on mouse B cells via membrane Ig and TLR-4 to activate the MAPK and NF-κB signaling pathways.

    PubMed

    Yang, Shu-fa; Zhuang, Tai-feng; Si, Yan-mei; Qi, Ke-yan; Zhao, Juan

    2015-03-01

    This study aimed to characterize the immunopotentiating effects and immune receptors for Coriolus versicolor mushroom polysaccharides (CVP), a Chinese medicinal fungus that exerts anti-tumor activities by enhancing host immunity. Proliferation assays were used to determine whether CVP could activate splenocytes. Flow cytometry analysis and IgM and IgG detection were used to characterize CVP-binding cells. Immune receptors were analyzed in immunoprecipitation and western blot assays. The downstream signaling pathways were identified by western blotting or immunostaining. CVP significantly stimulated the proliferation of mouse splenocytes. Fluorescence-labeled CVP (fl-CVP) selectively stained mouse B cells, but not T cells. CVP induced the production of IgM and IgG1 with or without exogenous IL-4. Membrane Ig (B cell antigen-receptor, BCR) was identified as a CVP-binding protein in immunoprecipitation and western blot experiments. CVP-induced B cell proliferation could be significantly inhibited by anti-mouse immunoglobulin (Ig) blocking antibody (Fab) or in cells from TLR4-mutant mice (C3H/HeJ). Phosphorylation of ERK-1/2 and p38 MAPK were clearly increased in a time-dependent manner, as was the nuclear translocation of the cytosolic NF-κB p65 subunit after CVP stimulation. Together, we demonstrate that CVP can bind and induce B cell activation using membrane Ig and TLR-4 as potential immune receptors. CVP activates mouse B cells through the MAPK and NF-κB signaling pathways.

  11. Speciation of mercury in mine waste: case study of abandoned and active gold mine sites at the Bibiani-Anwiaso-Bekwai area of South Western Ghana.

    PubMed

    Nartey, V K; Klake, R K; Doamekpor, L K; Sarpong-Kumankomah, S

    2012-12-01

    Speciation determines toxicity, transport pathways and residence time of a metal in different compartments of the environment. This study investigated the speciation of mercury in soils, derived from sites known for dumping of mine wastes in the Bibiani-Anwiaso-Bekwai district, a gold mining community of the Western Region of Ghana. Soil samples were taken from the surface; depths of 20, 40 and 60 cm from mine waste at both abandoned and active mine sites. Each sample was analysed for total mercury, organic mercury and elemental mercury. After sample treatment, digestion and reduction with stannous chloride (SnCl(2)), total mercury content was determined using the Inductively Coupled Plasma-Optical Emissions Spectrometer (ICP-OES). Organic mercury content was determined employing a differential technique after disposing of elemental mercury by heating. Total mercury content in samples ranged from 0.067 to 0.876 mg/kg for surface soils. The same soil of depths 20, 40 and 60 cm had total mercury from 0.102 to 1.066, 0.037 to 4.037 and 0.191 to 4.998 mg/kg, respectively. For organic mercury, concentrations range from 0.012 to 0.260 mg/kg for surface soil. Soil depths of 20, 40 and 60 cm had organic mercury concentrations from 0.016 to 0.653, 0.041 to 1.093 and 0.101 to 2.546 mg/kg respectively. Elemental mercury concentrations in surface soils, soils at depths of 20, 40 and 60 cm ranged from 0.043 to 0.780; 0.017 to 0.749; 0.014 to 2.944 and 0.009 to 2.452 mg/kg respectively. Among the sites studied, only galamsey tailings (GM) showed a trend of increasing total mercury level with increasing depth. For the other sites, trends were not defined. There has been no defined trend for elemental mercury with depth at any of the sampling sites. Just as with total mercury, it was only GM that showed an increasing trend of organic mercury concentration with depth.

  12. Active tectonics of the western tethyan himalaya above the underthrusting indian plate: The upper sutlej river basin as a pull-apart structure

    NASA Astrophysics Data System (ADS)

    Ni, James; Barazangi, Muawia

    1985-03-01

    Fault-bounded blocks and structural elements were mapped in the eastern Ladakh-Spiti and upper Sutlej River Basin located within the Tethyan Himalaya and to the southwest of the Karakorum fault zone mainly using LANDSAT Multispectral Scanner (MSS) band 5, band 7 (near-infrared) images with detailed analysis of smaller areas by interactive digital processing of false color images, and Returned Beam Vidicon (RBV) imagery in conjunction with available topographical, geological and seismological data. For the first time the Leo Pargil Horst and other nearby fault-bounded blocks located at the northwestern end of the upper Setlej River Basin were clearly revealed on the LANDSAT color composites. Shallow crustal seismicity is systematically related to the NNE-trending and N-trending normal faults of the Leo Pargil and nearby regions. Some of the aftershocks of the Kinnaur earthquake of January 19,1975 ( Ms = 6.8), appear to be associated with movement along the NNE-trending westbound fault of the Leo Pargil Horst and the nearby Kaurik-Chango fault. The main shock, however, is teleseismically located at about 30 km to the northwest of the Kaurik-Chango fault. Fault plane solutions of the main shock and two aftershocks indicate a large component of normal faulting. In map view, the upper Sutlej River Basin has an approximately rhomboidal shape and is located to the southwest of the Karakorum fault system. We suggest that this basin is a pull-apart between the NW-SE oriented, right-lateral, strike-slip Karakorum fault system and the high-angle faults near the southern boundary of the Tethyan Himalaya. The Leo Pargil Horst is the northwestern bounding fault block of this pull-apart. The active tectonic features in this part of the Tethyan Himalaya appear to reflect right-shear within the crust, and this is probably a consequence of oblique underthrusting of the Indian continental plate beneath the western Himalaya and southwestern Tibet during the Neogene and Quaternary

  13. Active crustal deformation across the Basin and Range province, western United States, measured with the Global Positioning System, 1992-2002

    NASA Astrophysics Data System (ADS)

    Hammond, W.; Thatcher, W.

    2003-04-01

    The Basin and Range province of the western United States is a region of active tectonic extension and dextral shear, accommodating roughly 25% of the motion between non-deforming North America (NA) and the Pacific Plate (PA). The orientation of dextral shear is consistent with that of NA/PA relative plate motion, suggesting that this high elevation interior province is an important part of the plate boundary system. We present an analysis of Global Positioning System (GPS) data collected from 1992 to 2002. An 800 km long network of campaign-style geodetic benchmarks extends from east of the Wasatch fault zone (WFZ) in central Utah to west of the Genoa fault zone and Lake Tahoe in the northern Sierra Nevada mountains. From the new data collected in September 2002 and from data collected in 1992, 1996, and 1998, velocities have been estimated at 92 GPS sites, nearly double the number previously presented by Thatcher et al. [1999]. This new data reduces the uncertainty in site velocities and increases the spatial detail compared to earlier results, and now allows resolution of distinct domains in the tensor strain rate field. To process the data we use the GIPSY/OASIS and Quasi-Observation Combination Analysis (Dong et al. [1998]) software packages and incorporate data from continuously recording GPS stations in California and Nevada. The results show that most of the approximately 12 mm/yr of Sierra Nevada block motion is accommodated by right lateral shear and extensional deformation concentrated in the westernmost 200 km of Nevada, in the vicinity of the Walker Lane (WL). A lesser amount of velocity variation (roughly 3 mm/yr) is localized at the easternmost edge of the network, in the vicinity of the Wasatch Fault Zone (WFZ). Estimates of tensor strain rates show transitions in the style of deformation. Near the WFZ only uniaxial, roughly east-west extension can be resolved. Between longitude -112 and -117.5 no deformation is resolvable. Near longitude -118, in

  14. Speciation of mercury in mine waste: case study of abandoned and active gold mine sites at the Bibiani-Anwiaso-Bekwai area of South Western Ghana.

    PubMed

    Nartey, V K; Klake, R K; Doamekpor, L K; Sarpong-Kumankomah, S

    2012-12-01

    Speciation determines toxicity, transport pathways and residence time of a metal in different compartments of the environment. This study investigated the speciation of mercury in soils, derived from sites known for dumping of mine wastes in the Bibiani-Anwiaso-Bekwai district, a gold mining community of the Western Region of Ghana. Soil samples were taken from the surface; depths of 20, 40 and 60 cm from mine waste at both abandoned and active mine sites. Each sample was analysed for total mercury, organic mercury and elemental mercury. After sample treatment, digestion and reduction with stannous chloride (SnCl(2)), total mercury content was determined using the Inductively Coupled Plasma-Optical Emissions Spectrometer (ICP-OES). Organic mercury content was determined employing a differential technique after disposing of elemental mercury by heating. Total mercury content in samples ranged from 0.067 to 0.876 mg/kg for surface soils. The same soil of depths 20, 40 and 60 cm had total mercury from 0.102 to 1.066, 0.037 to 4.037 and 0.191 to 4.998 mg/kg, respectively. For organic mercury, concentrations range from 0.012 to 0.260 mg/kg for surface soil. Soil depths of 20, 40 and 60 cm had organic mercury concentrations from 0.016 to 0.653, 0.041 to 1.093 and 0.101 to 2.546 mg/kg respectively. Elemental mercury concentrations in surface soils, soils at depths of 20, 40 and 60 cm ranged from 0.043 to 0.780; 0.017 to 0.749; 0.014 to 2.944 and 0.009 to 2.452 mg/kg respectively. Among the sites studied, only galamsey tailings (GM) showed a trend of increasing total mercury level with increasing depth. For the other sites, trends were not defined. There has been no defined trend for elemental mercury with depth at any of the sampling sites. Just as with total mercury, it was only GM that showed an increasing trend of organic mercury concentration with depth. PMID:22270596

  15. Crustal-scale shear zones recording 400 m.y. of tectonic activity in the North Caribou greenstone belt, western Superior Province of Canada

    NASA Astrophysics Data System (ADS)

    Kalbfleisch, Netasha

    A series of crustal-scale shear zones demarcates the northern and eastern margins of the North Caribou greenstone belt (NCGB), proximal to a Mesoarchean terrane boundary in the core of the western Superior Province of Canada. The dominant deformation produced a pervasive steeply dipping fabric that trends broadly parallel to the doubly arcuate shape of the belt and was responsible for tight folding the banded iron formation host to Goldcorp's prolific gold deposit at Musselwhite mine. The shear zones in the North Caribou greenstone belt are of particular interest because of their ability to channel hydrothermal fluids with the potential to bear ore and cause alteration of the middle to shallow crust. Shear zones are commonly reactivated during subsequent tectonism, but exhibit a consistent and dominant dextral shear sense across the belt; fabric-forming micas and chlorite are generally Mg-rich. Although garnets samples from within the shear zones are dominantly almandine, they possess variable geochemical trends (HREEs of >2 orders of magnitude) and can be syn-, intra-, or post-tectonic in origin. In situ geochronological analysis of zircon (U-Pb) and monazite (total-Pb) in high strain rocks in and around the NCGB, interpreted in light of in situ geochemical analysis of garnet and fabric-forming micas and chlorite, reveals four relatively discrete events that span 400 million years. Metamorphism of the mid-crust was coeval with magmatism during docking of the Island Lake domain at c. 2.86 Ga and subsequent terrane accretion at the north and south margins of the North Caribou Superterrane from c. 2.75 to 2.71 Ga. Transpressive shear at c. 2.60 to 2.56 Ga and late re-activation of shear zones at c. 2.44 Ga produced a steeply-dipping pervasive fabric, and channeled fluids for late crystallization of garnet and monazite recorded in the Markop Lake deformation zone. These observations implicate a horizontal tectonic model similar to the modern eastern Pacific plate

  16. Female mice lacking active nadph-oxidase enzymes are protected against “western diet”--induced obesity and metabolic syndrome

    Technology Transfer Automated Retrieval System (TEKTRAN)

    NADPH oxidase (Nox) enzymes have been implicated in regulation of adipocyte differentiation and inflammation in a variety of tissues. We examined the effects of feeding AIN-93G or a “Western diet” (WD) (45% fat, 0.5% cholesterol) on development of obesity and “metabolic syndrome” in wild type (WT) m...

  17. Summary of Activities and Evaluation of Western Alabama Cooperative Title IV Project: A Desegregation Program Occasioned by and Relative to Teacher Crossover.

    ERIC Educational Resources Information Center

    Tuscaloosa County Board of Education, Tuscaloosa, AL.

    The Western Alabama Cooperative Project attempted to develop better self-concepts, communication skills, classroom management techniques, and knowledge of multi-racial educational methods and materials among teachers and educators; the intent was to facilitate school desegregation. The program consisted of lectures and discussion groups. The…

  18. Da0324, an inhibitor of nuclear factor-κB activation, demonstrates selective antitumor activity on human gastric cancer cells

    PubMed Central

    Jin, Rong; Xia, Yiqun; Chen, Qiuxiang; Li, Wulan; Chen, Dahui; Ye, Hui; Zhao, Chengguang; Du, Xiaojing; Shi, Dengjian; Wu, Jianzhang; Liang, Guang

    2016-01-01

    Background The transcription factor nuclear factor-κB (NF-κB) is constitutively activated in a variety of human cancers, including gastric cancer. NF-κB inhibitors that selectively kill cancer cells are urgently needed for cancer treatment. Curcumin is a potent inhibitor of NF-κB activation. Unfortunately, the therapeutic potential of curcumin is limited by its relatively low potency and poor cellular bioavailability. In this study, we presented a novel NF-κB inhibitor named Da0324, a synthetic asymmetric mono-carbonyl analog of curcumin. The purpose of this study is to research the expression of NF-κB in gastric cancer and the antitumor activity and mechanism of Da0324 on human gastric cancer cells. Methods The expressions between gastric cancer tissues/cells and normal gastric tissues/cells of NF-κB were evaluated by Western blot. The inhibition viability of compounds on human gastric cancer cell lines SGC-7901, BGC-823, MGC-803, and normal gastric mucosa epithelial cell line GES-1 was assessed with the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. Absorption spectrum method and high-performance liquid chromatography method detected the stability of the compound in vitro. The compound-induced changes of inducible NF-κB activation in the SGC-7901 and BGC-823 cells were examined by Western blot analysis and immunofluorescence methods. The antitumor activity of compound was performed by clonogenic assay, matrigel invasion assay, flow cytometric analysis, Western blot analysis, and Hoechst 33258 staining assay. Results High levels of p65 were found in gastric cancer tissues and cells. Da0324 displayed higher growth inhibition against several types of gastric cancer cell lines and showed relatively low toxicity to GES-1. Moreover, Da0324 was more stable than curcumin in vitro. Western blot analysis and immunofluorescence methods showed that Da0324 blocked NF-κB activation. In addition, Da0324 significantly inhibited tumor proliferation

  19. 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.

  20. Examination of Proteins Bound to Nascent DNA in Mammalian Cells Using BrdU-ChIP-Slot-Western Technique.

    PubMed

    Bhaskara, Srividya

    2016-01-14

    Histone deacetylases 1 and 2 (HDAC1,2) localize to the sites of DNA replication. In the previous study, using a selective inhibitor and a genetic knockdown system, we showed novel functions for HDAC1,2 in replication fork progression and nascent chromatin maintenance in mammalian cells. Additionally, we used a BrdU-ChIP-Slot-Western technique that combines chromatin immunoprecipitation (ChIP) of bromo-deoxyuridine (BrdU)-labeled DNA with slot blot and Western analyses to quantitatively measure proteins or histone modification associated with nascent DNA. Actively dividing cells were treated with HDAC1,2 selective inhibitor or transfected with siRNAs against Hdac1 and Hdac2 and then newly synthesized DNA was labeled with the thymidine analog bromodeoxyuridine (BrdU). The BrdU labeling was done at a time point when there was no significant cell cycle arrest or apoptosis due to the loss of HDAC1,2 functions. Following labeling of cells with BrdU, chromatin immunoprecipitation (ChIP) of histone acetylation marks or the chromatin-remodeler was performed with specific antibodies. BrdU-labeled input DNA and the immunoprecipitated (or ChIPed) DNA was then spotted onto a membrane using the slot blot technique and immobilized using UV. The amount of nascent DNA in each slot was then quantitatively assessed using Western analysis with an anti-BrdU antibody. The effect of loss of HDAC1,2 functions on the levels of newly synthesized DNA-associated histone acetylation marks and chromatin remodeler was then determined by normalizing the BrdU-ChIP signal obtained from the treated samples to the control samples.

  1. Examination of Proteins Bound to Nascent DNA in Mammalian Cells Using BrdU-ChIP-Slot-Western Technique.

    PubMed

    Bhaskara, Srividya

    2016-01-01

    Histone deacetylases 1 and 2 (HDAC1,2) localize to the sites of DNA replication. In the previous study, using a selective inhibitor and a genetic knockdown system, we showed novel functions for HDAC1,2 in replication fork progression and nascent chromatin maintenance in mammalian cells. Additionally, we used a BrdU-ChIP-Slot-Western technique that combines chromatin immunoprecipitation (ChIP) of bromo-deoxyuridine (BrdU)-labeled DNA with slot blot and Western analyses to quantitatively measure proteins or histone modification associated with nascent DNA. Actively dividing cells were treated with HDAC1,2 selective inhibitor or transfected with siRNAs against Hdac1 and Hdac2 and then newly synthesized DNA was labeled with the thymidine analog bromodeoxyuridine (BrdU). The BrdU labeling was done at a time point when there was no significant cell cycle arrest or apoptosis due to the loss of HDAC1,2 functions. Following labeling of cells with BrdU, chromatin immunoprecipitation (ChIP) of histone acetylation marks or the chromatin-remodeler was performed with specific antibodies. BrdU-labeled input DNA and the immunoprecipitated (or ChIPed) DNA was then spotted onto a membrane using the slot blot technique and immobilized using UV. The amount of nascent DNA in each slot was then quantitatively assessed using Western analysis with an anti-BrdU antibody. The effect of loss of HDAC1,2 functions on the levels of newly synthesized DNA-associated histone acetylation marks and chromatin remodeler was then determined by normalizing the BrdU-ChIP signal obtained from the treated samples to the control samples. PMID:26863264

  2. Detection of Rickettsia in Rhipicephalus sanguineus ticks and Ctenocephalides felis fleas from southeastern Tunisia by reverse line blot assay.

    PubMed

    Khrouf, Fatma; M'Ghirbi, Youmna; Znazen, Abir; Ben Jemaa, Mounir; Hammami, Adnene; Bouattour, Ali

    2014-01-01

    Ticks (n = 663) and fleas (n = 470) collected from domestic animals from southeastern Tunisia were screened for Rickettsia infection using reverse line blot assay. Evidence of spotted fever group Rickettsia was obtained. We detected Rickettsia felis in fleas, Rickettsia massiliae Bar 29 and the Rickettsia conorii Israeli spotted fever strain in ticks, and Rickettsia conorii subsp. conorii and Rickettsia spp. in both arthropods. The sensitivity of the adopted technique allowed the identification of a new association between fleas and R. conorii subsp. conorii species. The presence of these vector-borne Rickettsia infections should be considered when diagnosing this disease in humans in Tunisia.

  3. Interspecies sharing of two distinct nonstructural protein 1 alleles among human and animal rotaviruses as revealed by dot blot hybridization.

    PubMed

    Fujiwara, Y; Nakagomi, O

    1997-10-01

    The distribution of the nonstructural protein 1 (NSP1) alleles from human strain AU-1 and canine strain K9 among rotaviruses of human, feline, canine, bovine, and simian origin was studied by a dot blot hybridization assay. Human and feline strains belonging to the AU-1 genogroup had the same NSP1 allele, while canine and feline strains belonging to the canine-feline genogroup shared another NSP1 allele. This canine-feline NSP1 allele had a significant level of homology with the NSP1 of rhesus rotavirus strain MMU18006. PMID:9316942

  4. Decrease in water activity due to fluid absent partial melting monitored with water content in biotite in the Western Adamello contact aureole (Italy)

    NASA Astrophysics Data System (ADS)

    Siron, Guillaume; Baumgartner, Lukas; Bouvier, Anne-Sophie; Vennemann, Torsten

    2016-04-01

    The fluorine and chlorine exchange on the hydroxyl site in micas is used to monitor changes in fluid composition (Munoz 1984). Most studies assume that the OH-site does not contain vaccancies, since the vast majority of studies use analytical techniques that does not allow to directly measure the OH- content of the mica. Nevertheless, studies have shown that significant amounts of O2- are present, and its concentration increases with temperature and titanium content. This feature was intrepreted as the consequence of a Ti-oxygen exchange in amphibolite and granulite facies rocks (Dyar et al. 1993, Cesare et al. 2008). Here, we present OH, F, Cl data for biotite from contact aureoles from biotite-schist to partially molten sillimanite-cordierite schists. OH-F-Cl content of biotites were analyzed using Secondary Ion Mass Spectrometry (SIMS), and major elements were analyzed by EMP. Samples were collected in the mid-crustal Western Adamello contact aureole (Italian Alps, Floess and Baumgartner, 2013). For that purpose we used biotite standards with water content constrained by Thermal Conversion Elemental Analyzer (TC/EA) see Bauer and Vennemann 2014, at a level of precision of 0.1-0.2 wt% (2SD). SIMS measurements typically have a precision of 0.1wt% (2SD), corresponding to the homogeneity of the internal standard at the SwissSIMS laboratory. OH- content decreases in samples with increasing peak temperature and Ti content (Ti range for biotites of 0.15-0.42 p.f.u for all samples). Nevertheless, within each individual sample, OH- is not a function of Ti. Ti variations are about 0.4 p.f.u., which is ten times the analytical uncertainty of the EMP analysis (0.004 p.f.u., 1SD). Water content is constant within analytical precision for each sample. The mean of OH- measurements is 3.41 p.f.u. in biotite and garnet grade samples, whereas those of samples in the partially molten zone have values of 3.27 p.f.u. We do not see any correlation with XMg or F and Cl. Hence, we

  5. WESTERN ECOLOGY DIVISION - GENERAL INFORMATION SHEET

    EPA Science Inventory

    abstract for flyer - general information The Western Ecology Division (WED), part of EPAs National Health and Environmental Effects Research Laboratory, provides information to EPA offices and regions nationwide to improve understanding of how human activities affect estuarine,...

  6. RT-PCR and Northern blot analysis in search for a putative Paramecium beta-adrenergic receptor.

    PubMed

    Płatek, A; Wiejak, J; Wyroba, E

    1999-01-01

    RT-PCR and Northern blot analysis were performed in order to search for a putative beta-adrenergic receptor (beta-AR) in Paramecium using several beta2-adrenergic-specific molecular probes. Under strictly defined RT-PCR conditions DNA species of expected molecular size about 360 bp were generated with the primers corresponding to the universal mammalian beta2-AR sequence tagged sites (located within the 4th and the 6th transmembrane regions of the receptor). This RT-PCR product hybridized in Southern blot analysis with the oligonucleotide probe designed to the highly conservative beta2-AR region involved in G-proteins interaction and located within the amplified region. Northern hybridization was performed on Paramecium total RNA and mRNA with human beta2-AR cDNA and two oligonucleotide probes: the first included Phe 290 involved in agonist binding (Strader et al., 1995) and the second was the backward RT-PCR primer. All these probes revealed the presence of about 2 kb mRNA which is consistent with the size of beta2-AR transcripts found in higher eukaryotes.

  7. Homocysteine Induces Heme Oxygenase-1 Expression via Transcription Factor Nrf2 Activation in HepG2 Cells

    PubMed Central

    Mani, Monireh; Golmohammadi, Taghi; Khaghani, Shahnaz; Zamani, Zahra; Azadmanesh, Kayhan; Meshkani, Reza; Pasalar, Parvin

    2013-01-01

    Background: Elevated level of plasma homocysteine has been related to various diseases. Patients with hyperhomocysteinemia can develop hepatic steatosis and fibrosis. We hypothesized that oxidative stress induced by homocysteine might play an important role in pathogenesis of liver injury. Also, the cellular response designed to combat oxidative stress is primarily controlled by the transcription factor Nrf2, a principal inducer of anti-oxidant and phase II-related genes. Methods: HepG2 cells were treated with homocysteine in different time periods. Glutathione content was measured by flowcytometry. Using electrophoretic mobility shift assay (EMSA) and Western-blotting, anti-oxidant response element (ARE)-binding activity of Nrf2 for heme ocygenase-1 (HO-1) was demonstrated. Real time RT-PCR and Western-blotting were performed to evaluate whether homocysteine was able to induce mRNA and protein expression of HO-1. Results: The role of Nrf2 in cellular response to homocysteine is substantiated by the following observations in HepG2 cells exposed to homocysteine (i) Western-blotting revealed that Nrf2 is strongly stabilized and became detectable in nuclear extracts. (ii) EMSA demonstrated increased binding of Nrf2 to oligomers containing HO-1 promoter-specific ARE-binding site. (iii) Real time RT-PCR and Western-blotting revealed increased mRNA and protein expression of inducible gene HO-1 after treatment with homocysteine. Conclusion: Data presented in the current study provide direct evidence that the immediate cellular response to oxidative stress provoked by homocysteine is orchestrated mainly by the Nrf2-ARE pathway. Therefore, induction of Nrf2-ARE-dependent expression of HO-1 could be a therapeutic option for hepatic cells damage induced by homocysteine. PMID:23567851

  8. 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

  9. RTG resource book for western states and provinces: Final proceedings

    SciTech Connect

    1994-12-31

    The Western Interstate Energy Board held a workshop and liaison activities among western states, provinces, and utilities on the formation of Regional Transmission Groups (RTGs). Purpose of the activities was to examine the policy implications for western states and provinces in the formation of RTGs in the West, the implications for western ratepayers and utilities of the RTG formation and potential impacts of RTGs on the western electricity system. The workshop contributed to fulfilling the transmission access and competition objectives of Title VII of the Energy Policy Act of 1992.

  10. What Is Western Civilization?

    ERIC Educational Resources Information Center

    Birken, Lawrence

    1992-01-01

    Discusses opposing tendencies in the interpretation of Western Civilization. Describes the expanded definition that includes Byzantine and Islamic cultures as heirs of the Greco-Roman cultures. Suggests that a limited definition of Western culture will facilitate a problems approach, emphasize diversity among cultures, and integrate the classical…

  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. Detection of human cytomegalovirus by slot-blot hybridization assay employing oligo-primed /sup 32/P-labelled probe

    SciTech Connect

    Agha, S.A.; Coleman, J.C.; Selwyn, S.; Mahmound, L.A.; Abd-Elaal, A.M.; Archard, L.C.

    1988-12-01

    A /sup 32/P-labelled Hind III-0 DNA fragment (nine Kilobases; Kb) from human cytomegalovirus AD-169 (HCMV) was used in slot-blot hybridization assay for the detection of HCMV in clinical samples. The results obtained with DNA hybridization assay (DNA HA) were compared with virus isolation using conventional tube cell culture (CTC) and centrifugation vial culture (CVC), immunofluorescence (IF), and complement fixation test (CFT). Of 15 CTC-positive samples, 13 were positive with DNA HA (sensitivity 86.7%). Also, 14 additional samples were DNA HA-positive but CTC-negative. CVC and/or IF confirmed the diagnosis in nine of 14; the remaining five samples were from three patients who showed fourfold rising antibody titre by CFT. Although DNA HA using /sup 32/P-labelled probes is relatively cumbersome and expensive, it is a valuable test for quantitation of viral shedding in patients with HCMV infections who may benefit from antiviral therapy.

  13. Amplification with molecular beacon primers and reverse line blotting for the detection and typing of human papillomaviruses.

    PubMed

    Jordens, J Z; Lanham, S; Pickett, M A; Amarasekara, S; Abeywickrema, I; Watt, P J

    2000-09-01

    A novel method for the detection and typing of human papillomavirus (HPV) was developed using molecular beacon primers. The method is based on the use of HPV-specific primers containing a hairpin loop structure in which fluorescent donor and quencher groups are held in close proximity such that fluorescence is quenched. Amplification of the target sequence results in the opening of the loop and the resulting fluorescence can be detected on a sequence detector system (SDS) 7700 (Applied Biosystems), as used for TaqMan assays. Fluorescent amplicons were identified on the SDS 7700 and then typed by a single hybridisation with specific probes immobilised in lines on a nylon membrane and detected on a fluorescent scanner. This novel beacon primer method compared well with conventional PCR for cervical scrape specimens. The combination of the beacon primer method and reverse line blotting should enable large-scale population studies of HPV infection.

  14. 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.

  15. Identifying active interplate and intraplate fault zones in the western Caribbean plate from seismic reflection data and the significance of the Pedro Bank fault zone in the tectonic history of the Nicaraguan Rise

    NASA Astrophysics Data System (ADS)

    Ott, B.; Mann, P.

    2015-12-01

    The offshore Nicaraguan Rise in the western Caribbean Sea is an approximately 500,000 km2 area of Precambrian to Late Cretaceous tectonic terranes that have been assembled during the Late Cretaceous formation of the Caribbean plate and include: 1) the Chortis block, a continental fragment; 2) the Great Arc of the Caribbean, a deformed Cretaceous arc, and 3) the Caribbean large igneous province formed in late Cretaceous time. Middle Eocene to Recent eastward motion of the Caribbean plate has been largely controlled by strike-slip faulting along the northern Caribbean plate boundary zone that bounds the northern margin of the Nicaraguan Rise. These faults reactivate older rift structures near the island of Jamaica and form the transtensional basins of the Honduran Borderlands near Honduras. Recent GPS studies suggest that small amount of intraplate motion within the current margin of error of GPS measurements (1-3 mm/yr) may occur within the center of the western Caribbean plate at the Pedro Bank fault zone and Hess Escarpment. This study uses a database of over 54,000 km of modern and vintage 2D seismic data, combined with earthquake data and results from previous GPS studies to define the active areas of inter- and intraplate fault zones in the western Caribbean. Intraplate deformation occurs along the 700-km-long Pedro Bank fault zone that traverses the center of the Nicaraguan Rise and reactivates the paleo suture zone between the Great Arc of the Caribbean and the Caribbean large igneous province. The Pedro Bank fault zone also drives active extension at the 200-km-long San Andres rift along the southwest margin of the Nicaraguan Rise. Influence of the Cocos Ridge indentor may be contributing to reactivation of faulting along the southwesternmost, active segment of the Hess Escarpment.

  16. Inhibition of the aryl hydrocarbon receptor prevents Western diet-induced obesity. Model for AHR activation by kynurenine via oxidized-LDL, TLR2/4, TGFβ, and IDO1.

    PubMed

    Moyer, Benjamin J; Rojas, Itzel Y; Kerley-Hamilton, Joanna S; Hazlett, Haley F; Nemani, Krishnamurthy V; Trask, Heidi W; West, Rachel J; Lupien, Leslie E; Collins, Alan J; Ringelberg, Carol S; Gimi, Barjor; Kinlaw, William B; Tomlinson, Craig R

    2016-06-01

    Obesity is an increasingly urgent global problem, yet, little is known about its causes and less is known how obesity can be effectively treated. We showed previously that the aryl hydrocarbon receptor (AHR) plays a role in the regulation of body mass in mice fed Western diet. The AHR is a ligand-activated nuclear receptor that regulates genes involved in a number of biological pathways, including xenobiotic metabolism and T cell polarization. This study was an investigation into whether inhibition of the AHR prevents Western diet-based obesity. Male C57Bl/6J mice were fed control and Western diets with and without the AHR antagonist α-naphthoflavone or CH-223191, and a mouse hepatocyte cell line was used to delineate relevant cellular pathways. Studies are presented showing that the AHR antagonists α-naphthoflavone and CH-223191 significantly reduce obesity and adiposity and ameliorates liver steatosis in male C57Bl/6J mice fed a Western diet. Mice deficient in the tryptophan metabolizing enzyme indoleamine 2,3-dioxygenase 1 (IDO1) were also resistant to obesity. Using an AHR-directed, luciferase-expressing mouse hepatocyte cell line, we show that the transforming growth factor β1 (TGFβ1) signaling pathway via PI3K and NF-κB and the toll-like receptor 2/4 (TLR2/4) signaling pathway stimulated by oxidized low-density lipoproteins via NF-κB, each induce luciferase expression; however, TLR2/4 signaling was significantly reduced by inhibition of IDO1. At physiological levels, kynurenine but not kynurenic acid (both tryptophan metabolites and known AHR agonists) activated AHR-directed luciferase expression. We propose a hepatocyte-based model, in which kynurenine production is increased by enhanced IDO1 activity stimulated by TGFβ1 and TLR2/4 signaling, via PI3K and NF-κB, to perpetuate a cycle of AHR activation to cause obesity; and inhibition of the AHR, in turn, blocks the cycle's output to prevent obesity. The AHR with its broad ligand binding specificity

  17. Violence the Western way.

    PubMed

    Roth, B E

    1997-10-01

    Despite the quiet revolution in response to changing conceptualizations of gender in psychoanalysis, the Western has remained the domain of aggressive phallic masculinity. The iconic imagery of the Western, when combined with its narrative trajectory, is used to tell stories of violent encounters between men. The acceptance of the genre, and its duplication by other cultures and film makers, indicates that the Westerns' imagery and moral solutions tap into some basic deep structures of anxiety and pleasure in violence between men. As long as societies require subtle sublimations of aggressive and violent drives, it is likely that men will seek imaginary regressive experiences to discharge frustrations.

  18. The role of active and ancient geothermal processes in the generation, migration, and entrapment of oil in the basin and Range Province, western USA. Final technical report

    SciTech Connect

    Hulen, J.B.; Collister, J.W.; Curtiss, D.K.

    1997-06-01

    The Basin and Range (B&R) physiographic province of the western USA is famous not only for its geothermal and precious-metal wealth, but also for its thirteen oil fields, small but in some cases highly productive. The Grant Canyon field in Railroad Valley, for example, for years boasted production of more than 6000 barrels of oil (BO) per day from just two wells; aggregate current production from the Blackburn field in Pine Valley commonly exceeds 1000 BO per day. These two and several other Nevada oil fields are unusually hot at reservoir depth--up to 130{degrees}C at depths as shallow as 1.1 km, up to three times the value expected from the prevailing regional geothermal gradient.

  19. Exploratory analysis of extremely low tropical cyclone activity during the late-season of 2010 and 1998 over the western North Pacific and the South China Sea

    NASA Astrophysics Data System (ADS)

    Zhao, Haikun; Chu, Pao-Shin; Hsu, Pang-Chi; Murakami, Hiroyuki

    2014-12-01

    This study attempts to understand why the tropical cyclone (TC) frequency over the western North Pacific and the South China Sea was so low in 2010 and 1998 even though a strong La Niña signal occurred in both years. We found that the TC frequency during the late-season (October to December), not in the peak season (July to September), makes 2010 a record low year; the next lowest year is 1998. Specifically, four TCs were observed over the South China Sea (SCS) in the late-season of 1998, but no TCs occurred over the SCS in the same season during 2010. The genesis potential index is used to help diagnose changes in environmental conditions for TC genesis frequency. Results indicate that the decreased low-level vorticity makes the largest contribution to the decreased TC formation over the SCS. The second largest contribution comes from the enhanced vertical wind shear, with relatively small contributions from the negative anomaly in potential intensity and reduction in midlevel relative humidity. These observational results are consistent with numerical simulations using a state of the art model from the Meteorological Research Institute (MRI-AGCM 3.2 Model). Numerical experiments show that the unfavorable conditions for sharply decreased TC formation during the late-season over the SCS in 2010 mainly results from the sea surface temperature anomaly over the western North Pacific basin. This effect is partly offset by the sea surface temperature anomaly in the South Indian Ocean and Northern Indian Ocean basins.

  20. Diagnostic use of the lymphocyte transformation test-memory lymphocyte immunostimulation assay in confirming active Lyme borreliosis in clinically and serologically ambiguous cases.

    PubMed

    Puri, Basant K; Segal, Daniel Rm; Monro, Jean A

    2014-01-01

    The aim of this study was to carry out an independent evaluation of the proposition that the lymphocyte transformation test-memory lymphocyte immunostimulation assay (LTT-MELISA) may be diagnostically useful in the confirmation of active Lyme borreliosis in clinically and serologically ambiguous cases. Blood samples from 54 patients consecutively presenting to a British center with clinical suspicion of Lyme borreliosis were tested for this disease by immunoglobulin M (IgM) and immunoglobulin G (IgG) Western blots and by LTT-MELISA. Forty-five of these patients had Western blot results which were negative for both IgM and IgG by the criteria of the Centers for Disease Control and Prevention (CDC); of these patients, 19 (42%) were LTT-MELISA-positive. Two of the patients who had IgM positive results by the CDC criteria were LTT-MELISA-negative. It is concluded that, for putative European-acquired Lyme borreliosis infections, it would be sensible to carry out both the LTT-MELISA and Western blot assay. PMID:25664127

  1. Diagnostic use of the lymphocyte transformation test-memory lymphocyte immunostimulation assay in confirming active Lyme borreliosis in clinically and serologically ambiguous cases

    PubMed Central

    Puri, Basant K; Segal, Daniel RM; Monro, Jean A

    2014-01-01

    The aim of this study was to carry out an independent evaluation of the proposition that the lymphocyte transformation test-memory lymphocyte immunostimulation assay (LTT-MELISA) may be diagnostically useful in the confirmation of active Lyme borreliosis in clinically and serologically ambiguous cases. Blood samples from 54 patients consecutively presenting to a British center with clinical suspicion of Lyme borreliosis were tested for this disease by immunoglobulin M (IgM) and immunoglobulin G (IgG) Western blots and by LTT-MELISA. Forty-five of these patients had Western blot results which were negative for both IgM and IgG by the criteria of the Centers for Disease Control and Prevention (CDC); of these patients, 19 (42%) were LTT-MELISA-positive. Two of the patients who had IgM positive results by the CDC criteria were LTT-MELISA-negative. It is concluded that, for putative European-acquired Lyme borreliosis infections, it would be sensible to carry out both the LTT-MELISA and Western blot assay. PMID:25664127

  2. 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.

  3. A Southern Blot Assay for Detection of Hepatitis B Virus Covalently Closed Circular DNA from Cell Cultures

    PubMed Central

    Cai, Dawei; Nie, Hui; Yan, Ran; Guo, Ju-Tao; Block, Timothy M.; Guo, Haitao

    2016-01-01

    Chronic hepatitis B remains a substantial public health burden affecting approximately 350 million people worldwide, causing cirrhosis and liver cancer, and about 1 million people die each year from hepatitis B and its complications. Hepatitis B is caused by hepatitis B virus (HBV) infection. As an essential component of the viral life cycle, HBV covalently closed circular DNA (cccDNA) is synthesized and maintained at low copy numbers in the nucleus of infected hepatocytes, and serves as the transcription template for all viral RNAs. Therefore, cccDNA is responsible for the establishment of viral infection and persistence. The presence and longevity of cccDNA may also explain the limitations of current antiviral therapy for hepatitis B. Thus, understanding the mechanisms underlying cccDNA formation and regulation is critical in understanding the HBV pathogenesis and finding a cure for hepatitis B. Here we describe a protocol for HBV cccDNA extraction and detection in detail. The procedure includes two major steps: (1) HBV cccDNA extraction by Hirt protein-free DNA extraction method and (2) HBV cccDNA detection by Southern blot analysis. The method is straightforward and reliable for cccDNA assay with cell culture samples, and it is useful for both HBV molecular biology and antiviral research. PMID:23821267

  4. [Establishment of a method for HLA-DRB genotyping in cord blood by reverse dot-blot hybridization technique].

    PubMed

    Huang, Yi-Ning; Liao, Can; Tang, Xue-Wei; Li, Yan; Xie, Xing-Mei; Zeng, Rui-Ping

    2002-04-01

    The HLA system was discovered by virtue of the fact that it was polymorphic. The impetus for its discovery was the search for polymorphic antigens to match for transplantation, by analogy with the human red cell blood groups. The most usually DNA method of HLA typing is sequence specific oligonucleotides (SSO) and PCR sequence specific primers (SSP). SSO technique is perfectly suited for analyzing large number of samples, it is not suitable for individual or small numbers. The SSP method is ideal for typing individual samples, but it is costly and requires high capacity thermal cycles for larger numbers of samples. To set up a simple, quick, cheap and high resolution DNA method, were collected sixty-three cord blood samples from Guangzhou Cord Blood Bank, got DNA from blood by the traditional guanidine hydrochloride distillation method. Each sample was simultaneously typed by SSOP, PCR-SSP and reverse dot-blot hybridization (RDB) methods. All of typed is success. The results of three DNA methods are consistent each other. 60 HLA-DRB1 alleles could be accurately distinguished with the RDB method. Our results show that RDB method is a simple, quick, cheap and high resolution method for HLA-DRB types. It can be used in any HLA typing.

  5. 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.

  6. Development of rapid, sensitive and non-radioactive tissue-blot diagnostic method for the detection of citrus greening.

    PubMed

    Nageswara-Rao, Madhugiri; Miyata, Shin-Ichi; Ghosh, Dilip; Irey, Mike; Garnsey, Stephen M; Gowda, Siddarame

    2013-01-01

    Citrus huanglongbing (HLB or citrus greening) is one of the most devastating diseases of citrus worldwide. The disease is caused by Gram-negative, phloem-limited α-proteobacterium, 'Candidatus Liberibacter asiaticus', vectored by the psyllid, Diaphorina citri Kuwayama. Citrus plants infected by the HLB bacterium may not show visible symptoms sometimes for years following infection and non-uniform distribution within the tree makes the detection of the pathogen very difficult. Efficient management of HLB disease requires rapid and sensitive detection early in the infection followed by eradication of the source of pathogen and the vector. The polymerase chain reaction (PCR) based method is most commonly employed for screening the infected/suspected HLB plants and psyllids. This is time consuming, cumbersome and not practical for screening large number of samples in the field. To overcome this, we developed a simple, sensitive, non-radioactive, tissue-blot diagnostic method for early detection and screening of HLB disease. Digoxigenin labeled molecular probes specific to 'Ca. L. asiaticus' nucleotide sequences have been developed and used for the detection of the pathogen of the HLB disease. The copy number of the target genes was also assessed using real-time PCR experiments and the optimized real-time PCR protocol allowed positive 'Ca. L. asiaticus' detection in citrus samples infected with 'Ca. L. asiaticus' bacterium.

  7. 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