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Sample records for activity western blotting

  1. 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...

  2. 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

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

    PubMed

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

    2013-09-17

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

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

    PubMed

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

    2011-06-01

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

  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. An automated rotisserie system for processing Western blots.

    PubMed

    Ma, P W K

    2002-01-01

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

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

    PubMed

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

    2007-08-01

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

  11. 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.

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

    PubMed Central

    2015-01-01

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

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

    PubMed

    Gerver, Rachel E; Herr, Amy E

    2014-11-01

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

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

    PubMed

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

    2014-03-01

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

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

    PubMed

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

    1999-09-01

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

  16. 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

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

    PubMed

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

    2016-01-01

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

  18. 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…

  19. 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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

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

    2014-10-01

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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed Central

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

    2000-01-01

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

  5. 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

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

    PubMed Central

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

    2016-01-01

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

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

    PubMed

    Gilda, Jennifer E; Gomes, Aldrin V

    2015-01-01

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

  8. 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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  11. 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…

  12. 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

  13. 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...

  14. 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…

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

    PubMed

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

    2016-09-01

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

  16. 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

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

    PubMed

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

    2016-03-01

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

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

    PubMed Central

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

    1990-01-01

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

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

    PubMed

    Carthew, James; Karakesisoglou, Iakowos

    2016-01-01

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

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

    NASA Astrophysics Data System (ADS)

    Farrell, Shawn O.; Farrell, Lynn E.

    1995-08-01

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

  1. 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

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

    PubMed

    Dangoudoubiyam, Sriveny; Kazacos, Kevin R

    2009-11-01

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

  3. 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

  4. 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

  5. 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

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

    PubMed Central

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

    1990-01-01

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

  7. 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

  8. 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

  9. 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

  10. 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

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

    PubMed

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

    2006-12-01

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

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

    PubMed Central

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

    2015-01-01

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

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

    PubMed

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

    2016-06-21

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

  14. 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.

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

    PubMed

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

    2005-01-01

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

  16. 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

  17. 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...

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

    PubMed Central

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

    1989-01-01

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

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

    PubMed

    Wiśniewski, Jacek R; Mann, Matthias

    2016-07-01

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

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

    PubMed

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

    2008-12-01

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

  1. 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

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

    PubMed Central

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

    2015-01-01

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

  3. 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

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

    PubMed Central

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

    2015-01-01

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

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

    PubMed

    Duggan, A E; Callard, I P

    2001-08-01

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

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

    PubMed Central

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

    2014-01-01

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

  7. 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

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

    PubMed Central

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

    1985-01-01

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

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

    PubMed

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

    2005-06-10

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

  10. 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

  11. 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

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

    PubMed

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

    2008-06-01

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

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

    PubMed

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

    2010-07-14

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

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

    PubMed

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

    2012-04-30

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

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

    PubMed Central

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

    2015-01-01

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

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

    PubMed Central

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

    2016-01-01

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

  17. 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

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

    PubMed Central

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

    2014-01-01

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

  19. 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)

  20. 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

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

    PubMed

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

    2016-07-01

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

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

    PubMed

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

    2009-08-26

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

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

    PubMed Central

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

    1992-01-01

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

  4. 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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  6. 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

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

    PubMed

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

    1999-09-01

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

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

    PubMed

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

    2010-11-01

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

  9. 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

  10. 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

  11. 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

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

    PubMed

    Alba, F J; Daban, J R

    1998-10-01

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

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

    PubMed

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

    2016-01-01

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

  14. 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

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

    PubMed

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

    2016-08-01

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

  16. 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

  17. 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

  18. 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

  19. 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

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

    PubMed

    Patrick, Weber; Hans, Steinhart; Angelika, Paschke

    2009-09-23

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

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

    PubMed

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

    2013-08-01

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

  2. 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

  3. 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. PMID:25036383

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

    PubMed

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

    2014-01-01

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

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

    PubMed

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

    2014-05-01

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

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

    PubMed

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

    2014-12-01

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

  8. 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

  9. BlotBase: a northern blot database.

    PubMed

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

    2008-12-31

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

  10. Identification of immunodominant VP1 linear epitope of enterovirus 71 (EV71) using synthetic peptides for detecting human anti-EV71 IgG antibodies in Western blots.

    PubMed

    Foo, D G W; Ang, R X; Alonso, S; Chow, V T K; Quak, S H; Poh, C L

    2008-03-01

    A major IgG-specific immunodominant VP1 linear epitope of enterovirus 71 (EV71) strain 41 (5865/SIN/00009), defined by the core sequence LEGTTNPNG, was identified by Pepscan analysis. Oligonucleotides corresponding to the amino-acid sequence of synthetic peptide SP32 were cloned and over-expressed in Escherichia coli as a recombinant glutathione-S-transferase (GST)-SP32 fusion protein. In ELISAs, this protein did not react with human anti-EV71 IgG antibodies, but there was significant immunoreactivity according to western blot analysis. The amino-acid sequence of SP32 was highly specific for detecting EV71 strains in western blot analysis, and showed no immunoreactivity with monoclonal antibodies raised against other enteroviruses, e.g., CA9 and Echo 6. PMID:18076666

  11. Southwestern Blotting Assay

    PubMed Central

    Jia, Yinshan; Nagore, Linda; Jarrett, Harry

    2016-01-01

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

  12. 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,…

  13. Genomic Southern blot analysis.

    PubMed

    Gebbie, Leigh

    2014-01-01

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

  14. 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

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

    PubMed Central

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

    2014-01-01

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

  16. Significance of quantitative enzyme-linked immunosorbent assay (ELISA) results in evaluation of three ELISAs and Western blot tests for detection of antibodies to human immunodeficiency virus in a high-risk population.

    PubMed Central

    Nishanian, P; Taylor, J M; Korns, E; Detels, R; Saah, A; Fahey, J L

    1987-01-01

    The characteristics of primary (first) tests with three enzyme-linked immunosorbent assay (ELISA) kits for human immunodeficiency virus (HIV) antibody were determined. The three ELISAs were performed on 3,229, 3,130, and 685 specimens from high-risk individuals using the Litton (LT; Litton Bionetics Laboratory Products, Charleston, S.C.), Dupont (DP; E. I. du Pont de Nemours & Co., Inc., Wilmington, Del.), and Genetic Systems (GS; Genetic Systems, Seattle, Wash.) kits, respectively. Evaluation was based on the distribution of quantitative test results (such as optical densities), a comparison with Western blot (WB) results, reproducibility of the tests, and identification of seroconverters. The performances of the GS and the DP kits were good by all four criteria and exceeded that of the LT kit. Primary ELISA-negative results were not always confirmed with repeat ELISA and by WB testing. The largest percentage of these unconfirmed negative test results came from samples with quantitative results in the fifth percentile nearest the cutoff. Thus, supplementary testing was indicated for samples with test results in this borderline negative range. Similarly, borderline positive primary ELISA results that were quantitatively nearest (fifth percentile) the cutoff value were more likely to be antibody negative on supplementary testing than samples with high antibody values. In this study, results of repeated tests by GS ELISA showed the least change from first test results. DP ELISA showed more unconfirmed primary positive test results, and LT ELISA showed more unconfirmed primary negative test results. Designation of a specimen with a single ELISA quantitative level near the cutoff value as positive or negative should be viewed with skepticism. A higher than normal proportion of specimens with high negative optical densities by GS ELISA (fifth percentile nearest the cutoff) and also negative by WB were found to be from individuals in the process of seroconversion. PMID

  17. Assessment of diagnostic accuracy of a commercial ELISA for the detection of Toxoplasma gondii infection in pigs compared with IFAT, TgSAG1-ELISA and Western blot, using a Bayesian latent class approach.

    PubMed

    Basso, Walter; Hartnack, Sonja; Pardini, Lais; Maksimov, Pavlo; Koudela, Bretislav; Venturini, Maria C; Schares, Gereon; Sidler, Xaver; Lewis, Fraser I; Deplazes, Peter

    2013-06-01

    Serological methods are the most commonly used diagnostic tools to detect Toxoplasma gondii infections in pigs. In the absence of a readily available 'gold standard', an estimation of diagnostic accuracy is difficult to assess. A commercial ELISA (PrioCHECK® Toxoplasma Ab porcine ELISA, Prionics, Schlieren, Switzerland) for the diagnosis of T. gondii infection in pigs was evaluated in naturally infected animals from two distinct populations; indoor and outdoor living animals. An assessment of diagnostic accuracy, using a Bayesian latent class approach with adjustment for within indoor and outdoor farm clustering using random effects, was performed. Tests used for comparison were: IFAT; ELISA using native affinity-purified P30 (SAG1) T. gondii tachyzoite surface antigen (TgSAG1-ELISA); and Western blot with T. gondii tachyzoites lysate. The data set comprised 297 pig serum samples across outdoor (n=149) and indoor (n=148) farms in Argentina. The estimated sensitivity and specificity for the commercial ELISA were 98.9% (95% credible interval: 96.2; 100) and 92.7% (95% credible interval: 87.7; 96.6), respectively. The analysis of sera and plasma from pigs (n=6) experimentally inoculated with 5,000 T. gondii oocysts revealed a pronounced antibody response beginning 2 weeks p.i. until the end of the observation period (11 weeks p.i.) in all animals. Meat juice obtained from inoculated animals after euthanasia also tested positive. These results suggest that the PrioCHECK® Toxoplasma Ab porcine ELISA may be a useful tool to perform serological diagnosis of T. gondii infections in pigs to control Toxoplasma infection in pigs and humans. PMID:23538054

  18. 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

  19. 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

  20. 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

  1. 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

  2. The impact of the Western Ghats on lightning activity on the western coast of India

    NASA Astrophysics Data System (ADS)

    Kamra, A. K.; Nair, A. A.

    2015-06-01

    The effect of the Western Ghats on the lightning activity across the west coast of India around the coastal metropolitan city of Mumbai during the 1998-2012 period is investigated using data from the Lightning Imaging Sensor (LIS) onboard the Tropical Rainfall Measuring Mission (TRMM) satellite. A land-sea contrast of an order of magnitude in the lightning activity is observed even in a small area across the western coast of India. The shape of a zone of high lightning activity formed almost parallel to the Western Ghats during the onset and withdrawal phases of monsoon, strongly suggests the effect of the Western Ghats in its formation. Seasonal variation of the lightning activity in this area and also in each of its four equal sectors (two each over the Arabian Sea and over land) is bi-annual with one peak each in the onset (May/June) and withdrawal months (September/October) of monsoon and a sharp dip to very low values during the monsoon months (July/August) of maximum seasonal rainfall. The lightning activity in each sector is found to increase over the 1998-2012 period. However, the increase in lightning activity over the sector containing Mumbai is found to be greater during the pre- and post-monsoon periods and smaller during the monsoon period as compared to an identical sector immediately south of it.

  3. 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…

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

    PubMed

    O'Sullivan, Jack; McMahon, Hilary E M

    2016-04-15

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

  5. False-Positive Results in a Recombinant Severe Acute Respiratory Syndrome-Associated Coronavirus (SARS-CoV) Nucleocapsid-Based Western Blot Assay Were Rectified by the Use of Two Subunits (S1 and S2) of Spike for Detection of Antibody to SARS-CoV

    PubMed Central

    Maache, Mimoun; Komurian-Pradel, Florence; Rajoharison, Alain; Perret, Magali; Berland, Jean-Luc; Pouzol, Stéphane; Bagnaud, Audrey; Duverger, Blandine; Xu, Jianguo; Osuna, Antonio; Paranhos-Baccalà, Glaucia

    2006-01-01

    To evaluate the reactivity of the recombinant proteins expressed in Escherichia coli strain BL21(DE3), a Western blot assay was performed by using a panel of 78 serum samples obtained, respectively, from convalescent-phase patients infected with severe acute respiratory syndrome-associated coronavirus (SARS-CoV) (30 samples) and from healthy donors (48 samples). As antigen for detection of SARS-CoV, the nucleocapsid protein (N) showed high sensitivity and strong reactivity with all samples from SARS-CoV patients and cross-reacted with all serum samples from healthy subjects, with either those obtained from China (10 samples) or those obtained from France (38 serum samples), giving then a significant rate of false positives. Specifically, our data indicated that the two subunits, S1 (residues 14 to 760) and S2 (residues 761 to 1190), resulted from the divided spike reacted with all samples from SARS-CoV patients and without any cross-reactivity with any of the healthy serum samples. Consequently, these data revealed the nonspecific nature of N protein in serodiagnosis of SARS-CoV compared with the S1 and S2, where the specificity is of 100%. Moreover, the reported results indicated that the use of one single protein as a detection antigen of SARS-CoV infection may lead to false-positive diagnosis. These may be rectified by using more than one protein for the serodiagnosis of SARS-CoV. PMID:16522785

  6. Cenozoic to active deformation in Western Yunnan (Myanmar China border)

    NASA Astrophysics Data System (ADS)

    Socquet, A.; Pubellier, M.

    2003-04-01

    The northward movement of India induces a right-lateral shear band from the Sunda trench to the easternmost Himalaya, where wrenching between India and Sunda plates, interfere with a clockwise flow of material around the Eastern Himalayan Syntaxis. We describe brittle and ductile deformation styles in Western Yunnan and Northern Myanmar, using field data and Landsat 7 imagery for Cenozoic structures as well as GPS and seismicity for active structures to unravel the Neogene to Present evolution. Western Yunnan is crossed by three continental-size ductile shear zones characterized by high mountain belts mainly composed of high-grade metamorphics and mylonitic rocks, and affected by active faulting. The easternmost metamorphic range, the Gaoligong Shan composed of verticalized foliated granites and mylonites is flattened westward and joins the Mogok metamorphic belt in Myanmar. East of the Gaoligong Shan, lie the Chong Shan and the Ailao / Diangcan Shan metamorphic ranges, which presents a vertical shistosity and a left-lateral motion. These three shear zones are separated by sedimentary fold-and-thrust-belts in the East, and , West of the Gaoligong, by Quaternary basins and volcanics. Preliminary results indicate that the Shan Scarp constituted the major strike-slip boundary between Indochina and India during Eocene to Miocene time, and accommodated deformation in right-lateral wrench. At the same time, the Ailao / Diangcan Shan and the Chong Shan zones were sheared left-laterally allowing the displacement toward the SE of Indochina block relative to south China. In the Miocene, ductile deformation migrated north along the Shan Scarp to the Mogok / Ruili metamorphic belt and the Gaoligong belt, dragging the Chong Shan right-laterally and superimposing a late right-lateral ductile deformation on its metamorphic rocks. The present-day relative motion between India and Sundaland, inferred from GPS processing, reaches 35 mm / yr in the Myanmar area. It is classically

  7. 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

  8. 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.

  9. 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.

  10. 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

  11. Active folded structures of the Western Caucasus (Sochi region)

    NASA Astrophysics Data System (ADS)

    Trikhunkov, Yaroslav; Zelenin, Egor

    2014-05-01

    The Western Caucasus as a margin segment of folded system of the Greater Caucasus was formed at the periphery of collision interaction of the Scythian Plate and the Transcaucasian Massif. The estimated age of the primary folded deformations of the initial surface of that territory ranges from the late Eocene to late Neogene. We have obtained new data on modern folded deformations of the anticlinal ridges, which prevail in Sochi region in the southern macroslope of the mountain system. Very similar Alek, Galitsinsky, Akhun, Nikolaevsky anticlinal ridges are uplifting in the main Caucasus direction (NW - SE) and are crossed by narrow antecedent river valleys. These ridges stand out contrasting to sinclinal depressions, where fluviatile accumulation prevails. At the intersection of the Mzymta river and the Galitsinsky anticlinal ridge a narrow Akhshtyr canyon with steep, 150 meters high slopes were formed. Downstream in the neighboring Akhshtyr synclinal depression the valley expands. Here the floodplain and two levels of terraces with the height of 20 - 30 and 50 - 60 m correspondingly were formed. The age of the first terrace was defined by archeologic data of V. Shchelinsky (2007) and by correlation with marine Black Sea Late Karangat terrace as a 135 - 90 ka (Eemian interglacial). The second terrace is apparently older and dates back to Middle Pleistocene. The field research and analysis of the elevations by ASTER GDEM allowed us to trace both terraces in the southern structural slope of the Galitsinsky ridge above the canyon, adjacent to the Akhshtyr depression, at the heights 70 and 110 m correspondingly. Alluvial deposits in outcrops of lower terrace (elongated pebbles, which look like modern alluvium of the Mzymta) were traced on the surface of the slope. Thereby, described fragments of the Mzymta terraces were uplifted above the level of the corresponding terraces in the synclinal depression as a result of dislocation on the slope of the actively uplifting

  12. 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,...

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

    PubMed

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

    2009-07-01

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

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

    PubMed

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

    2016-07-01

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

  15. GROUNDWATER QUALITY MONITORING OF WESTERN COAL STRIP MINING: PRELIMINARY DESIGNS FOR ACTIVE MINE SOURCES OF POLLUTION

    EPA Science Inventory

    Three potential pollution source categories have been identified for Western coal strip mines. These sources include mine stockpiles, mine waters, and miscellaneous active mine sources. TEMPO's stepwise monitoring methodology (Todd et al., 1976) is used to develop groundwater qua...

  16. 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. PMID:12158990

  17. Temperature increasing trend due to solar activity at Western Saudi

    NASA Astrophysics Data System (ADS)

    Almleaky, Y. M.; Sharaf, M. A.; Basurah, H. M.; Malawi, A. A.; Al-Mostafa, Z. A.

    The Sun influnce on climate has been discussed globaly by many authors and at different latitudes. In this article we will discuss this connection for the Kingdom of Saudi Arabia, which spans a large area, i.e. 16-32 North and 36-50 East. We started our invistigation in this paper by looking into the temperature at the Western coast of the Kingdom, namely Yenbo and Jeddah. In order to find the correlation between temperature and solar variations we employed one of the most relevant solar quentity, i.e. the solar cycle length. From our invistigations we found an increase in the temperature averages reaching up to 1.0 degree Celsius in certain cities since 1970. It is also found that the temperature increase is strongly correlated with the solar Cycle length, reaching up to 0.8 in some sites.

  18. ADA Compliance and the Accessibility of Physical Activity Facilities in Western Oregon.

    ERIC Educational Resources Information Center

    Cardinal, Bradley J.; Spaziani, Marc D.

    2003-01-01

    Investigated how compliant physical activity facilities in western Oregon were with regard to the Americans with Disabilities Act's mandate regarding accessibility of public facilities for all people. Data from direct observations and physical measurements at 50 physical activity facilities indicated that no facility was 100-percent compliant. The…

  19. 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

  20. Early Miocene Tectonic Activity in the western Ross Sea (Antarctica)

    NASA Astrophysics Data System (ADS)

    Sauli, C.; Sorlien, C. C.; Busetti, M.; Geletti, R.; De Santis, L.

    2012-12-01

    In the framework of the Rossmap Italian PNRA work objectives to compile extended and revised digital maps of the main unconformities in Ross Sea, Antarctica, much additional seismic reflection data, that were not available to previous ANTOSTRAT compilation, were incorporated into a new ROSSMAP interpretation. The correlation across almost all of Ross Sea, from DSDP Site 270 and Site 272 in Eastern Basin to northern Victoria Land Basin, of additional early Miocene and late Oligocene horizons that were not part of ANTOSTRAT allows interpretations to be made of fault activity and glacial erosion or deposition at a finer time resolution. New conclusions include that extensional or transtensional fault activity within the zone between Victoria Land Basin and Northern Basin, initiated by 23 Ma or earlier, and continued after 18 Ma. Steep parallel-striking faults in southern Victoria Land Basin display both reverse and normal separation of 17.5 Ma (from Cape Roberts Program-core 1) and post-16 Ma horizons, suggesting an important strike-slip component. This result may be compared with published papers that proposed post-17 Ma extension in southern Victoria Land Basin, 16-17 Ma extension in the AdareTrough, north of the Ross Sea continental shelf, but no Miocene extension affecting the Northern Basin (Granot et al., 2010). Thus, our evidence for extension through the early Miocene is significant to post-spreading tectonic models. Reference Granot R., Cande S. C., Stock J. M., Davey F. J. and Clayton R. W. (2010) Postspreading rifting in the Adare Basin, Antarctica: Regional tectonic consequences. Geochem. Geophys. Geosyst., 8, Q08005, doi:10.1029/2010GC003105.

  1. 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.

  2. 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

  3. 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

  4. 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.

  5. Quantitative autoradiography of dot blots using a microwell densitometer

    SciTech Connect

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

    1989-07-01

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

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

    EPA Science Inventory

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

  7. 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

  8. 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

  9. 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...

  10. 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.

  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. Seismic imaging of the geodynamic activity at the western Eger rift in central Europe

    NASA Astrophysics Data System (ADS)

    Mullick, N.; Buske, S.; Hrubcova, P.; Ruzek, B.; Shapiro, S.; Wigger, P.; Fischer, T.

    2015-04-01

    The western Eger rift at the Czech-German border in central Europe is an important geodynamically active area within the European Cenzoic rift system (ECRS) in the forelands of the Alps. Along with two other active areas of the ECRS, the French Massif Central and the east and west Eifel volcanic fields, it is characterized by numerous CO2-rich fluid emission points and frequent micro-seismicity. Existence of a plume(s) is indicated in the upper mantle which may be responsible for these observations. Here we reprocess a pre-existing deep seismic reflection profile '9HR' and interpret the subsurface structures as mapped by seismic reflectivity with previous findings, mainly from seismological and geochemical studies, to investigate the geodynamic activity in the subsurface. We find prominent hints of pathways which may allow magmatic fluids originating in the upper mantle to rise through the crust and cause the observed fluid emanations and earthquake activity.

  13. 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

  14. 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

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

    SciTech Connect

    Bodkin, D.K.

    1985-01-01

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

  16. Significant Centers of Tectonic Activity as Identified by Wrinkle Ridges for the Western Hemisphere of Mars

    NASA Technical Reports Server (NTRS)

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

    2000-01-01

    The western hemisphere region of Mars has been the site of numerous scientific investigations regarding its tectonic evolution. For this region of Mars, the dominant tectonic region is the Tharsis province. Tharsis is characterized by an enormous system of radiating grabens and a circumferential system of wrinkle ridges. Past investigations of grabens associated with Tharsis have identified specific centers of tectonic activity. A recent structural analysis of the western hemisphere region of Mars which includes the Tharsis region, utilized 25,000 structures to determine the history of local and regional centers of tectonic activity based primarily on the spatial and temporal relationships of extensional features. This investigation revealed that Tharsis is more structurally complex (heterogeneous) than has been previously identified: it consists of numerous regional and local centers of tectonic activity (some are more dominant and/or more long lived than others). Here we use the same approach as Anderson et al. to determine whether the centers of tectonic activity that formed the extensional features also contributed to wrinkle ridge (compressional) formation.

  17. 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.

  18. Multiple Seismic Array Observations for Tracing Deep Tremor Activity in Western Shikoku, Japan

    NASA Astrophysics Data System (ADS)

    Takeda, T.; Matsuzawa, T.; Shiomi, K.; Obara, K.

    2011-12-01

    Deep non-volcanic tremors become very active during episodic slow-slip events in western Japan and Cascadia. The episodic tremor and slow-slip events in western Shikoku, Japan, occur at a typical interval of 6 months. Recently, it has been reported that tremor migration activity is complex and shows different migrating directions depending on time scales (Ghosh et al., 2010). Such characteristics of tremor are important to understand the mechanism of tremor and the relationship between tremor and SSEs. However it is difficult to determine the location of tremors with high accuracy because tremors show faint signals and make the identification of P/S-wave arrivals difficult. Seismic array analysis is useful to evaluate tremor activity, especially to estimate the arrival direction of seismic energy (e.g. Ueno et al., 2010, Ghosh et al., 2010), as it can distinguish multiple tremor sources occurring simultaneously. Here, we have conducted seismic array observation and analyzed seismic data during tremor activity by applying the MUSIC method to trace tremor location and its migration in western Shikoku. We have installed five seismic arrays in western Shikoku since January 2011. One of the arrays contains 30 stations with 3-component seismometers with a natural frequency of 2 Hz (Type-L array). The array aperture size is 2 km and the mean interval between stations is approximately 200 m. Each of the other arrays (Type-S array) contains 9 seismic stations with the same type of seismometers of the Type-L array, and is deployed surrounding the Type-L array. The small array aperture size is 800 m and its mean station interval is approximately 150 m. All array stations have recorded continuous waveform data at a sampling of 200Hz. In May 2011, an episodic tremor and a short-term slip event occurred for the first time during the observation period. We could retrieve the array seismic data during the whole tremor episode. The analysis of data from the type-L array confirms

  19. 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.

  20. Atmospheric conditions associated with extreme fire activity in the Western Mediterranean region.

    PubMed

    Amraoui, Malik; Pereira, Mário G; DaCamara, Carlos C; Calado, Teresa J

    2015-08-15

    Active fire information provided by TERRA and AQUA instruments on-board sun-synchronous polar MODIS platform is used to describe fire activity in the Western Mediterranean and to identify and characterize the synoptic patterns of several meteorological fields associated with the occurrence of extreme fire activity episodes (EEs). The spatial distribution of the fire pixels during the period of 2003-2012 leads to the identification of two most affected sub-regions, namely the Northern and Western parts of the Iberian Peninsula (NWIP) and Northern Africa (NAFR). The temporal distribution of the fire pixels in these two sub-regions is characterized by: (i) high and non-concurrent inter- and intra-annual variability with maximum values during the summer of 2003 and 2005 in NWIP and 2007 and 2012 in NAFR; and, (ii) high intra-annual variability dominated by a prominent annual cycle with a main peak centred in August in both sub-regions and a less pronounced secondary peak in March only evident in NWIP region. The 34 EEs identified were grouped according to the location, period of occurrence and spatial configuration of the associated synoptic patterns into 3 clusters (NWIP-summer, NWIP-winter and NAFR-summer). Results from the composite analysis reveal similar fire weather conditions (statistically significant positive anomalies of air temperature and negative anomalies of air relative humidity) but associated with different circulation patterns at lower and mid-levels of the atmosphere associated with the occurrence of EEs in each cluster of the Western Mediterranean region. PMID:25889542

  1. Active rollback in the Gibraltar Arc: Evidences from CGPS data in the western Betic Cordillera

    NASA Astrophysics Data System (ADS)

    Gonzalez-Castillo, L.; Galindo-Zaldivar, J.; de Lacy, M. C.; Borque, M. J.; Martinez-Moreno, F. J.; García-Armenteros, J. A.; Gil, A. J.

    2015-11-01

    The Gibraltar Arc, located in the western Mediterranean Sea, is an arcuate Alpine orogen formed by the Betic and Rif Cordilleras, separated by the Alboran Sea. New continuous GPS data (2008-2013) obtained in the Topo-Iberia stations of the western Betic Cordillera allow us to improve the present-day deformation pattern related to active tectonics in this collision area between the Eurasian and African plates. These data indicate a very consistent westward motion of the Betic Cordillera with respect to the relatively stable Iberian Massif foreland. The displacement in the Betics increases toward the south and west, reaching maximum values in the Gibraltar Strait area (4.27 mm/yr in Ceuta, CEU1, and 4.06 mm/yr in San Fernando, SFER), then progressively decreasing toward the northwestern mountain front. The recent geological structures and seismicity evidence moderate deformation in a roughly NW-SE to WNW-ESE compressional stress setting in the mountain frontal areas, and moderate extension toward the internal part of the cordillera. The mountain front undergoes progressive development of folds affecting at least up to Pliocene deposits, with similar recent geological and geodetical rates. This folded strip helps to accommodate the active deformation with scarce associated seismicity. The displacement pattern is in agreement with the present-day clockwise rotation of the tectonic units in the northern branch of the Gibraltar Arc. Our data support that the westward emplacement of the Betic Cordillera continues to be active in a rollback tectonic scenario.

  2. 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.

  3. 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

  4. Geomorphological markers of faulting and neotectonic activity along the western Andean margin, northern Chile

    NASA Astrophysics Data System (ADS)

    Audin, Laurence; Herail, Gérard; Riquelme, Rodrigo; Darrozes, José; Martinod, Joseph; Font, Eric

    2003-12-01

    In the Atacama Desert, northern Chile, some ephemeral channels are developed in the Plio-Quaternary alluvial sequence that caps the Neogene Atacama Gravels Formation. Geomorphological studies and high-resolution digital elevation data (GPS) along a structural transect in the Central Depression are used to document modern growth history of subtle folding and faulting in the fore-arc region. Outcrop data of the most recent deposits are combined with observations of warped and faulted late Quaternary pediments, alluvial fans and terrace surfaces to propose unsuspected neotectonic processes on the western flank of the Domeyko Cordillera. Neotectonic process recognition is here based largely upon the interpretation of alluvial landforms, drainage organisation and evolution as the intermittent river network shows systematic patterns of course deflections, successive incisions or deposition processes as it encounters the fault scarps or folds in the superficial deposits. This area presents both N-S-trending active vertical faults in the topographically higher pampas, and N-S-trending active folding in the lower pampas. These faults seem to accommodate E-W extension and compression that could be related to uplift of the western Andean margin within a compressive context. Uplift may have taken place unevenly over the past few million years after the deposition of the superficial alluvial surfaces that cap the Neogene Atacama Gravels. Copyright

  5. Children's activities and their meanings for parents: a mixed-methods study in six Western cultures.

    PubMed

    Harkness, Sara; Zylicz, Piotr Olaf; Super, Charles M; Welles-Nyström, Barbara; Bermúdez, Moisés Ríos; Bonichini, Sabrina; Moscardino, Ughetta; Mavridis, Caroline Johnston

    2011-12-01

    Theoretical perspectives and research in sociology, anthropology, sociolinguistics, and cultural psychology converge in recognizing the significance of children's time spent in various activities, especially in the family context. Knowing how children's time is deployed, however, only gives us a partial answer to how children acquire competence; the other part must take into account the culturally constructed meanings of activities, from the perspective of those who organize and direct children's daily lives. In this article, we report on a study of children's routine daily activities and on the meanings that parents attribute to them in six Western middle-class cultural communities located in Italy, The Netherlands, Poland, Spain, Sweden, and the United States (N = 183). Using week-long time diaries kept by parents, we first demonstrate similarities as well as significant differences in children's daily routines across the cultural samples. We then present brief vignettes--"a day in the life" --of children from each sample. Parent interviews were coded for themes in the meanings attributed to various activities. Excerpts from parent interviews, focusing on four major activities (meals, family time, play, school- or developmentally related activities), are presented to illustrate how cultural meanings and themes are woven into parents' organization and understanding of their children's daily lives. The results of this mixed-method approach provide a more reliable and nuanced picture of children's and families' daily lives than could be derived from either method alone. PMID:22149041

  6. 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. PMID:26867274

  7. 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

  8. 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.

  9. 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...

  10. Utility of Western Blot Analysis for the Diagnosis of Cutaneous Leishmaniasis

    PubMed Central

    ASHRAFMANSOURI, Marzieh; SARKARI, Bahador; HATAM, Gholamreza; HABIBI, Parvaneh; ABDOLAHI KHABISI, Samaneh

    2015-01-01

    Background: Cutaneous leishmaniasis (CL) is a parasitic disease with a relatively wide distribution in different areas of the world, including Iran. The parasite is mainly diagnosed microscopically, but serological approaches might be useful for diagnosis as well. This study aimed to assess the efficacy of an immunoblotting system for serodiagnosis of cutaneous leishmaniasis in Iran. Methods: Sixty-one sera samples from parasitologically confirmed CL patients and 50 sera samples from healthy controls along with 50 sera sample from non-CL patients were collected. Native strain of Leishmania major was cultured in Schneider medium and soluble Leishmania antigens were prepared from amastigotes-like parasites. All of sera samples were evaluated by an immunoblotting system. Results: Components of 14 to 135 kDa were detectable by the sera of CL patients. From 61 sera of CL patients, 59 cases (96.7%) detected a 63 kDa subunit and 51 cases (83.6%) recognized a 32–35 kDa component. Among all subunits, the 63 kDa band showed the highest sensitivity (96.7%) and a 75 kDa band had the highest (98%) specificity. Conclusion: Immunoblotting has a satisfactory performance in diagnosis of CL and this test can be used, as an aid, for proper diagnosis of CL. PMID:26811727

  11. 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. PMID:26044007

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

    PubMed

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

    2016-01-01

    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. PMID:27341489

  13. Increasing eolian dust deposition in the western United States linked to human activity

    NASA Astrophysics Data System (ADS)

    Neff, J. C.; Ballantyne, A. P.; Farmer, G. L.; Mahowald, N. M.; Conroy, J. L.; Landry, C. C.; Overpeck, J. T.; Painter, T. H.; Lawrence, C. R.; Reynolds, R. L.

    2008-03-01

    Mineral aerosols from dust are an important influence on climate and on marine and terrestrial biogeochemical cycles. These aerosols are generated from wind erosion of surface soils. The amount of dust emission can therefore be affected by human activities that alter surface sediments. However, changes in regional- and global-scale dust fluxes following the rapid expansion of human populations and settlements over the past two centuries are not well understood. Here we determine the accumulation rates and geochemical properties of alpine lake sediments from the western interior United States for the past 5,000 years. We find that dust load levels increased by 500% above the late Holocene average following the increased western settlement of the United States during the nineteenth century. We suggest that the increased dust deposition is caused by the expansion of livestock grazing in the early twentieth century. The larger dust flux, which persists into the early twenty-first century, results in a more than fivefold increase in inputs of K, Mg, Ca, N and P to the alpine ecosystems, with implications for surface-water alkalinity, aquatic productivity and terrestrial nutrient cycling.

  14. An Overview of Recent Geostationary Fire Monitoring Activities and Applications in the Western Hemisphere

    NASA Astrophysics Data System (ADS)

    McRae, D. J.; Conard, S. G.; Ivanova, G. A.; Sukhinin, A. I.; Hao, W. M.; Koutzenogii, K. P.; Prins, E. M.; Schmidt, C. C.; Feltz, J. M.

    2002-05-01

    Over the past twenty years the international scientific research and environmental monitoring communities have recognized the vital role environmental satellites can play in detecting and monitoring active fires both regionally and around the globe for hazards applications and to better understand the extent and impact of biomass burning on the global environment. Both groups have stressed the importance of utilizing operational satellites to produce routine fire products and to ensure long-term stable records of fire activity for applications such as land-use/land cover change analyses and global climate change research. The current NOAA GOES system provides the unique opportunity to detect fires throughout the Western Hemisphere every half-hour from a series of nearly identical satellites for a period of 15+ years. This presentation will provide an overview of the GOES biomass burning monitoring program at UW-Madison Cooperative Institute for Meteorological Satellite Studies (CIMSS) with an emphasis on recent applications of the new GOES Wildfire Automated Biomass Burning Algorithm (WF_ABBA). For the past 8 years, CIMSS has utilized the GOES-8 imager to monitor biomass burning trends in South America. Since September 2000, CIMSS has been producing half-hourly fire products in real-time for most of the Western Hemisphere. The WF_ABBA half-hourly fire product is providing new insights into diurnal, spatial, seasonal and interannual fire dynamics in North, Central, and South America. In North America these products are utilized to detect and monitor wildfires in northerly and remote locations. In South America the diurnal GOES fire product is being used as an indicator of land-use and land-cover change and carbon dynamics along the borders between Brazil, Peru, and Bolivia. The Navy is assimilating the Wildfire ABBA fire product into the Navy Aerosol Analysis and Prediction System (NAAPS) to analyze and predict aerosol loading and transport as part of the NASA

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

    SciTech Connect

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

    1987-05-01

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

  16. 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

  17. 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

  18. 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.

  19. Higher Order Loess Terracettes and Related Ungulate Activities in Western Pottawattamie County, Iowa

    NASA Astrophysics Data System (ADS)

    Weihs, B. J.

    2009-12-01

    Terracettes are small, quasi-parallel, staircase-like, stepped landforms. They are generally less than a meter in tread width and riser height and as long as 300 meters, located transversely along slopes. Many theories purport to explain the mechanisms that cause them, including animal disturbance, soil creep, solifluction (gelifluction), slumping and rotational slippage, regolith control, vegetation control, subsidence, and anthropogenic or tectonic activities. This thesis was aimed at morphologically characterizing terracettes in the western Iowa Loess Hills, with an emphasis on further classifying the forms morphogenetically. Onsite observations suggest that terracettes in this area are highly associated with anthropogenically induced grazing of domestic ungulates such as cattle, horses, and sheep, as well as deer. A new class of terracette (higher order or mega-terracette) is proposed that characterizes (and differentiates) the study area forms based on size, in that the study area contains terracettes that are an order above those discussed by other authors (sub-meter). This new addition to the current standard (tear terracettes-risers bare of vegetation, and normal terracettes-normal to the slope), as suggested by Anderson (1972), will add to the local understanding of the forms, and especially to the theory of polygenesis in which terracettes can result from a host of processes. This new class of terracette (mega) is higher magnitude, and directly related to ungulate activities such as geophagy, soil transport (from hooves), compaction, smearing, pawing, and wallowing (dust bathing) as well as the effects of variable soil moisture on erosion of the forms used by the animals. High magnitude terracettes in western Pottawattamie County, Iowa

  20. 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

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. Hydrologic vulnerability of western US rangelands in the wake of woodland encroachment and increasing wildfire activity

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 a...

  6. 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

  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. 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.

  10. 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.

  11. 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

  12. 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

  13. 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

  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. The April 2007 earthquake swarm near Lake Trichonis and implications for active tectonics in western Greece

    NASA Astrophysics Data System (ADS)

    Kiratzi, A.; Sokos, E.; Ganas, A.; Tselentis, A.; Benetatos, C.; Roumelioti, Z.; Serpetsidaki, A.; Andriopoulos, G.; Galanis, O.; Petrou, P.

    2008-06-01

    We investigate the properties of the April 2007 earthquake swarm (Mw 5.2) which occurred at the vicinity of Lake Trichonis (western Greece). First we relocated the earthquakes, using P- and S-wave arrivals to the stations of the Hellenic Unified Seismic Network (HUSN), and then we applied moment tensor inversion to regional broad-band waveforms to obtain the focal mechanisms of the strongest events of the 2007 swarm. The relocated epicentres, cluster along the eastern banks of the lake, and follow a distinct NNW-ESE trend. The previous strong sequence close to Lake Trichonis occurred in June-December 1975. We applied teleseismic body waveform inversion, to obtain the focal mechanism solution of the strongest earthquake of this sequence, i.e. the 31 December 1975 (Mw 6.0) event. Our results indicate that: a) the 31 December 1975 Mw 6.0 event was produced by a NW-SE normal fault, dipping to the NE, with considerable sinistral strike-slip component; we relocated its epicentre: i) using phase data reported to ISC and its coordinates are 38.486°N, 21.661°E; ii) using the available macroseismic data, and the coordinates of the macroseismic epicentre are 38.49°N, 21.63°E, close to the strongly affected village of Kato Makrinou; b) the earthquakes of the 2007 swarm indicate a NNW-SSE strike for the activated main structure, parallel to the eastern banks of Lake Trichonis, dipping to the NE and characterized by mainly normal faulting, occasionally combined with sinistral strike-slip component. The 2007 earthquake swarm did not rupture the well documented E-W striking Trichonis normal fault that bounds the southern flank of the lake, but on the contrary it is due to rupture of a NW-SE normal fault that strikes at a ˜ 45° angle to the Trichonis fault. The left-lateral component of faulting is mapped for the first time to the north of the Gulf of Patras which was previously regarded as the boundary for strike-slip motions in western Greece. This result signifies the

  16. 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

  17. 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

  18. 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.

  19. Shallow Seismic Reflection Study of Recently Active Fault Scarps, Mina Deflection, Western Nevada

    NASA Astrophysics Data System (ADS)

    Black, R. A.; Christie, M.; Tsoflias, G. P.; Stockli, D. F.

    2006-12-01

    During the spring and summer of 2006 University of Kansas geophysics students and faculty acquired shallow, high resolution seismic reflection data over actively deforming alluvial fans developing across the Emmigrant Peak (in Fish Lake Valley) and Queen Valley Faults in western Nevada. These normal faults represent a portion of the transition from the right-lateral deformation associated with the Walker Lane/Eastern California Shear Zone to the normal and left-lateral faulting of the Mina Deflection. Data were gathered over areas of recent high resolution geological mapping and limited trenching by KU students. An extensive GPR data grid was also acquired. The GPR results are reported in Christie, et al., 2006. The seismic data gathered in the spring included both walkaway tests and a short CMP test line. These data indicated that a very near-surface P-wave to S-wave conversion was taking place and that very high quality S-wave reflections were probably dominating shot records to over one second in time. CMP lines acquired during the summer utilized a 144 channel networked Geode system, single 28 hz geophones, and a 30.06 downhole rifle source. Receiver spacing was 0.5 m, source spacing 1.0m and CMP bin spacings were 0.25m for all lines. Surveying was performed using an RTK system which was also used to develop a concurrent high resolution DEM. A dip line of over 400m and a strike line over 100m in length were shot across the active fan scarp in Fish Lake Valley. Data processing is still underway. However, preliminary interpretation of common-offset gathers and brute stacks indicates very complex faulting and detailed stratigraphic information to depths of over 125m. Depth of information was actually limited by the 1024ms recording time. Several west-dipping normal faults downstep towards the basin. East-dipping antithetic normal faulting is extensive. Several distinctive stratigraphic packages are bound by the faults and apparent unconformitites. A CMP dip line

  20. Hydrothermal Petroleum in Active Continental Rift: Lake Chapala, Western Mexico, Initial Results.

    NASA Astrophysics Data System (ADS)

    Zarate-del Valle, P. F.; Simoneit, B. R.; Ramirez-Sanchez, H. U.

    2003-12-01

    Lake Chapala in western Mexico is located partially in the Citala Rift, which belongs to the well-known neotectonic Jalisco continental triple junction. The region is characterized by active volcanism (Ceboruco, Volcan de Fuego), tectonic (1995 earthquake, M=8, 40-50 mm to SW) and hydrothermal (San Juan Cosala & Villa Corona spas and La Calera sinter deposit) activities. Hydrothermal petroleum has been described in active continental rift (East African Rift) and marine spreading zones (Guaymas Basin, Gulf of California). In 1868 the Mexican local press reported that manifestations of bitumen were appearing in front of the Columba Cap on the mid south shore of Lake Chapala. This bitumen is linked to the lake bottom and when the water level decreases sufficiently it is possible to access these tar bodies as islands. Because of these manifestations the Mexican oil company (PEMEX) drilled an exploration well (2,348m) at Tizapan El Alto without success. Hydrothermal activity is evident in the tar island zone as three in-shore thermal springs (26.8 m depth, 48.5° C, pH 7.8 and oriented N-S). The preliminary analyses by GC-MS of the tar from these islands indicate hydrothermal petroleum derived from lake sedimentary organic matter, generated at low temperatures (150° -200° C). The tars contain no n-alkanes, no PAH or other aromatics, but a major UCM of branched and cyclic hydrocarbons and mature biomarkers derived from lacustrine biota. The biomarkers consist of mainly 17α (H),21β (H)-hopanes ranging from C27 to C34 (no C28), gammacerane, tricyclic terpanes (C20-C26), carotane and its cracking products, and drimanes (C14-C16). The biomarker composition indicates an organic matter source from bacteria and algae, typical of lacustrine ecosystems. 14C dating of samples from two tar islands yielded ages exceeding 40 kyrs, i.e., old carbon from hydrothermal/tectonic remobilization of bitumen from deeper horizons to the surface. The occurrence of hydrothermal petroleum in

  1. 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.

  2. 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

  3. 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

  4. 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.

  5. 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

  6. 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

  7. 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

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

    DOE PAGESBeta

    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

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

    PubMed

    Zhao, Z; Lim, L; Manser, E

    2001-07-01

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

  10. 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

  11. 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.

  12. Geothermal activity at continental rift Citala, Western Mexico, where Lake Chapala is emplaced: past and present

    NASA Astrophysics Data System (ADS)

    Zárate-del Valle, P. F.

    2003-04-01

    Lake Chapala is a tectonic lake developed on a continental rift named Citala (CRC) which belongs to a tectonically active zone in Western Mexico: the so-called Jalisco continental triple junction. Fossil sinter deposit, thermal spring, hydrothermal (hy) petroleum manifestation and hy alteration halo characterized the Lake Chapala basin. On the SE shore, outcrops a carbonate deposit named ``La Calera" (LC) which consists of a carbonate fossil sinter that measures 2 km in E-W direction and 600 m in N-S direction and overlays andesitic rock. With a thickness of approximately 5 m and a roughly horizontal attitude, the LC is characterized by a two-fold structure: when massive, it is colored in yellow brownish and grey and elsewhere it shows a pseudo-brecciated structure and when banded, yellow and dark millimetre alternated bands can be seen. The LC is marked by vuggy porosity and silica (quartz and chalcedony) vein lets. Under microscope a pseudo-micritic texture is observed; vugs coated by iron oxides, are filled with calcite, and/or quartz, chalcedony and clay minerals. Six samples of LC were analysed (LODC-UParis VI) for their stable isotopes (δ18O and δ13C): From δ13C{PDB} values we have two sets of data: -8.03 to -8.69 ppm that means a no contribution of organic carbon (oc) and -0.35 to -0.75 ppm meaning an important contribution of oc; from δ18O{PDB} values: -8.5 to -10.27 ppm we deduced a deposit in meteoric water with a temperature deposition higher than the surface. The CRC is characterized also by the presence of hydrothermal petroleum (hp): Inside the Chapala and ˜2 km from SE shore (Los Arcos) there are some small spots made of hp which look like islands (<3-4 m^2) linked to the bottom of the lake which consist of solid bitumen. Thermal springs (ths) occur both inside and outside the lake Chapala: the water in out-shore ths is of carbonate type (69^oC; ˜ 240 mg L-1 [HCO_3]^-; with one exception: the ths at the San Juan Cosalá spa (N shore), which is

  13. Snow Water Equivalent distribution in the Western Alps: the activities in Aosta Valley

    NASA Astrophysics Data System (ADS)

    Morra di Cella, Umberto; Cremonese, Edoardo; Diotri, Fabrizio; Pogliotti, Paolo; Galvagno, Marta

    2010-05-01

    Snow can be seen as a water reservoir; it plays an important role in the hydrological cycle and climatology of the Earth. Its importance is enhanced in mountainous region where yearly streamflow is strongly controlled by snowmelt dynamics. Climate Change can influence precipitation distribution and snow cover persistence and thus can negatively affect water availability. The knowledge of the amount of water stored in the snow, its spatial distribution and its temporal evolution dynamics, is an issue of increasing importance. The Aosta Valley is a region in the western Italian Alps with a continental climate which can be strongly affected by a reduction of water availabilty. For these reasons, from 2006, the Environmental Protection Agency of Aosta Valley (ARPA Valle d'Aosta) is developping modelling activities aiming to monitor Snow Water Equivalent (SWE) distribution at regional scale (3000 Km2). To estimate SWE distribution we need to know the extent of the snow covered area (SCA) and how snow height (SH) and snow density (SD) are distributed in space. Because of the specific spectral reflectance of snow, SCA can be discriminated from snow-free areas using optical remote sensing methods. The MODIS standard snow-cover products (MOD10A2 maximum snow extent - 8 days composite) can be freely downloaded from the web and their spatial resolution (500 m) is well suited for the application at regional scale. Snow height data can be obtained from automatic measurements (ultrasonic distance sensors) done by the regional meteorological station networks or by manual measurements of snow depth, while snow density has to be manually measured in snow pits. It follows that SH dataset are often more data-rich than SD ones. Snow height and snow density spatial distribution is modelled using multiple regression models between the snow parameters and many morphological variables (e.g. elevation, slope, incoming solar radiation, topographical parameters, ...). Regression models are

  14. 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.

  15. 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...

  16. Insecticidal activity of monoterpenoids to western corn rootworm (Coleoptera: Chrysomelidae), twospotted spider mite (Acari: Tetranychidae), and house fly (Diptera: Muscidae).

    PubMed

    Lee, S; Tsao, R; Peterson, C; Coats, J R

    1997-08-01

    Acute toxicities of 34 naturally occurring monoterpenoids were evaluated against 3 important arthropod pest species; the larva of the western corn rootworm, Diabrotica virgifera virgifera LeConte; the adult of the twospotted spider mite. Tetranychus urticae Koch; and the adult house fly. Musca domestica L. Potential larvicidal or acaricidal activities of each monoterpenoid were determined by topical application, leaf-dip method, soil bioassay, and greenhouse pot tests. Phytotoxicity was also tested on a corn plant. Citronellic acid and thymol were the most topically toxic against the house fly, and citronellol and thujone were the most effective on the western corn rootworm. Most of the monoterpenoids were lethal to the twospotted spider mite at high concentrations; carvomenthenol and terpinen-4-ol were especially effective. A wide range of monoterpenoids showed some larvicidal activity against the western corn rootworm in the soil bioassay. Perillaldehyde, the most toxic (LC50 = 3 micrograms/g) in soil, was only 1/3 as toxic as carbofuran, a commercial soil insecticide (LC50 = 1 microgram/g). Selected monoterpenoids also effectively protected corn roots from attack by the western corn rootworm larvae under greenhouse conditions. alpha-Terpineol was the best monoterpenoid in the greenhouse pot test. The acute toxicity of monoterpenoids was low relative to conventional insecticides. Some monoterpenoids were phytotoxic to corn roots and leaves. l-Carvone was the most phytotoxic, whereas pulegone was the safest. The results with thymyl ethyl ether, one of the synthetic derivatives of thymol, showed a potential of derivatization to reduce monoterpenoid phytotoxicity. PMID:9260540

  17. Trends in water market activity and price in the western United States

    NASA Astrophysics Data System (ADS)

    Brown, Thomas C.

    2006-09-01

    Over 2000 water market transactions that occurred in the western United States from 1990 to 2003 were examined to learn who sold to whom and for what purpose, how much water was involved, and how much it sold for. The transactions show that much more water changes hands via leases than via sales of water rights. Public agencies and irrigators are the most common lessors, with lessees being fairly evenly distributed across types of buyers. However, with water rights sales, irrigators are by far the most common sellers and municipalities the most common buyers. Across the West in general, the number of leases has been rising in recent years, as have their prices. The prices of water right sales have also been rising, but the number of sales has not. The price of water is highly variable both within and between western states, reflecting the localized nature of the factors that affect water prices.

  18. 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, X.; Mickley, L. J.; Logan, J. A.

    2010-12-01

    We estimate future wildfire activity over the western United States during the mid-21st century (2046-2065), based on ensemble results from 16 models following the IPCC A1B scenario. Regression models and a parameterization model of area burned (AB) are developed for the projection. The ecoregion-based regression models consider the meteorological impacts from both the current and previous years on the regional AB; the correlation coefficients between the predicted and observed annual AB range from 0.59 to 0.85 during 1981-2000 for six ecoregions. The parameterization model calculates the effects of temperature, precipitation, and relative humidity on daily AB; the predicted monthly total AB over the western US is highly correlated with observations with a normal (logarithmic) correlation coefficient of 0.73 (0.90) for 300 months (1980-2004). The meteorological fields under the A1B scenario from 15 IPCC models and a general circulation model (NASA/GISS GCM 3) are applied to the fire models to estimate AB during 2046-2065. We calculate that the annual AB will increase by 21~124% (regression models) and 40~198% (parameterization) over six ecoregions in the western US at midcentury relative to present day; the length of fire season will extend by 11.1% for the warmer and drier climate. We also examine the impact of the changing climate on the fuel load with a dynamic vegetation model LPJ driven by future meteorology from the GISS GCM 3; it shows that the fuel load over western US experiences small changes because of the short time period from 2000 to 2050. Based on the projected fuel load and the AB calculations, we estimate that the annual total biomass burning over western US will increase by 68% (regression models) or as much as 161% (parameterization). We further investigate the impact of the changing fire emission on the carbonaceous aerosol concentrations over the western United States in the mid-21st century using the chemistry transport model GEOS-Chem driven

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

    PubMed Central

    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 (HBPSW) and SPM (HBPSPM) 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 HBPSW from the surface to 500 m was comparable between the locations, whereas HBPSPM 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

  20. 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

  1. 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.

  2. 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.

  3. Latent outflow activity for western Tharsis, Mars: Significant flood record exposed

    USGS Publications Warehouse

    Dohm, J.M.; Anderson, R.C.; Baker, V.R.; Ferris, J.C.; Rudd, L.P.; Hare, T.M.; Rice, J. W., Jr.; Casavant, R.R.; Strom, R.G.; Zimbelman, J.R.; Scott, D.H.

    2001-01-01

    Observations permitted by the newly acquired Mars Observer Laser Altimeter data have revealed a system of gigantic valleys northwest of the huge Martian shield volcano, Arsia Mons, in the western hemisphere of Mars (northwestern slope valleys (NSVs)). These features, which generally correspond spatially to gravity lows, are obscured by veneers of materials including volcanic lava flows, air fall deposits, and eolian materials. Geologic investigations of the Tharsis region suggest that the system of gigantic valleys predates the construction of Arsia Mons and its extensive associated lava flows of mainly late Hesperian and Amazonian age and coincides stratigraphically with the early development of the outflow channels that debouch into Chryse Planitia. Similar to the previously identified outflow channels, which issued tremendous volumes of water into topographic lows such as Chryse Planitia, the NSVs potentially represent flooding of immense magnitude and, as such, a source of water for a northern plains ocean.

  4. 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.

  5. 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).

  6. Active deformation in the inner western Alps inferred from comparison between 1972-classical and 1996-GPS geodetic surveys

    NASA Astrophysics Data System (ADS)

    Sue, Christian; Martinod, Joseph; Tricart, Pierre; Thouvenot, François; Gamond, Jean-François; Fréchet, Julien; Marinier, Delphine; Glot, Jean-Paul; Grasso, Jean-Robert

    2000-04-01

    Eighteen geodetic points surveyed in 1972 by the French National Geographic Institute (IGN) were remeasured by GPS in 1996 in the Briançonnais and Piémont Zones, east of the Pelvoux massif (French Western Alps). A displacement vector set was determined for the two surveys' common points. Calculations of the strain-rate tensors associated with 15 triangular cells of the network have been performed. Only four of them show a strain rate significant at a 95% level of confidence. These data suggest an E-W extension of about 2-4 mm/yr between the western and eastern part of the network (Pelvoux external crystalline massif and Queyras blueschists, respectively) associated with N-S shortening. This active deformation agrees with neotectonic and seismotectonic data. The measured tectonic motion seems to be distributed throughout the central part of the Briançonnais zone, where the seismic activity is concentrated. The local seismicity has been precisely surveyed since 1989. It is moderate ( Ml<4.7) and no larger earthquake occured in the 1972-1989 period. The seismic deformation of the 1972-1996 period, extrapolated from the 1989-1996 local seismicity, accounts for less than 10% of the geodetic deformation. Thus, aseismic processes accommodated more than 90% of the observed deformation during this period. This could correspond to accumulation of elastic strain on locked faults, creep on faults or plastic deformation in a large crustal volume.

  7. 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

  8. 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

  9. 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.

  10. In Vitro Antiplasmodial Activity and Cytotoxicity of Extracts of Selected Medicinal Plants Used by Traditional Healers of Western Cameroon

    PubMed Central

    Zofou, Denis; Tene, Mathieu; Ngemenya, Moses N.; Tane, Pierre; Titanji, Vincent P. K.

    2011-01-01

    Medicinal plants play a key role in malaria control in Africa, especially in remote areas where health facilities are limited. In order to assess their acclaimed potentials, eleven extracts were prepared from seven selected plants commonly used in Western Cameroon, and tested both for their antiplasmodial activity and cytotoxicity. The antiplasmodial activity was assessed using Lactate Dehydrogenase Assay (pLDH) and the cytotoxicity estimated on LLC-MK2 monkey kidney epithelial cells. Seven extracts from five different plants were significantly active, with very weak or no cytotoxicity. The Dacryodes edulis leaves showed the highest activity (IC50 of 6.45 μg/mL on 3D7 and 8.2 μg/mL on DD2) followed by the leaves of Vernonia amygdalina (IC50 of 8.72 and 11.27 μg/mL on 3D7 and DD2 resp.) and roots of V. amygdalina (IC50 of 8.72 μg/mL on 3D7), Coula edulis leaves (IC50 of 13.80 μg/mL and 5.79 μg/mL on 3D7 and DD2 resp.), Eucalyptus globulus leaves (IC50 of 16.80 μg/mL and 26.45 μg/mL on 3D7 and DD2) and Cuviera longiflora stem bark (IC50 of 20.24 μg/mL and 13.91 μg/mL on 3D7 and DD2). These findings justify the use of five of the seven plants in malaria treatment by traditional healers of Western Cameroon. PMID:22312569

  11. 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

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

    PubMed Central

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

    1999-01-01

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

  13. 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.

  14. 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

  15. 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

  16. 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

  17. 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.

  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. PMID:27441230

  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. Investigation of the relationships between seismic activities and radon level in western Turkey.

    PubMed

    Tarakçı, M; Harmanşah, C; Saç, M M; İçhedef, M

    2014-01-01

    The distribution of radon activity is determined from pre-earthquake data. Analysis using Normal, Gamma, Weibull and Rayleigh distributions indicates that the variation of radon levels in seismically active regions is best described by a normal distribution. It was observed that radon levels would change in compressive fault lines prior to earthquake. Besides that it tended to increase before the earthquake and then decrease towards the time of earthquake occurrences. PMID:24215813

  1. 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.

  2. 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

  3. Western Canada

    SciTech Connect

    Hay, P.W.; Robertson, D.C.

    1981-10-01

    In 1980, a third successive all-time drilling record was set in western Canada, with 8865 wells being drilled, up 20% since 1979. Exploratory drilling increased 30%, to 3744 wells, and development drilling increased 14%, to 5121 wells. The exploratory success rate increased to 66% in 1980, based on 1017 oil discoveries and 1463 gas discoveries. The development success rate increased marginally to 89%, with 1774 oil discoveries and 2778 gas discoveries. Average well depth increased in all four western provinces, and total land sales reached the record $1 billion mark in Alberta and a record $78 million in Saskatchewan. British Columbia land sales declined slightly to $181 million. Alberta drilling activity continued in the deeper portions of the Alberta basin and foothills, with major gas discoveries at Hanlan, Big Mountain, Blackstone, and Elmworth. Significant oil discoveries were made in the West Pembina Nisku pinnacle reefs, in the Upper Devonian at Del Bonita and Eaglesham, and in the Lower Cretaceous glauconite river channels in southern Alberta between Countess and Grand Forks. British Columbia successes occurred as the Elmworth Deep Basin play spilled over into British Columbia with gas discoveries at Tupper and Steeprock. Gas finds were also made at West Sierra and Murray. The Arctic Islands continued to yield the largest discoveries. Two major successes occurred in the Beaufort Sea, in an oil and gas discovery by Esso at Issungnak and a reentry oil discovery by Dome at Tarsuit. However, 1980 will especially be remembered for the introduction of the federal government's National Energy Program during October, with new taxes on revenue, lower than expected wellhead price increases, and major emphasis on increasing Canadian ownership and self-sufficiency. Industry and provincial government reaction was highly critical, and a major downturn in exploration is expected in western Canada in 1981. 3 figures, 8 tables.

  4. Slope failures and stability analysis of shallow water prodeltas in the active margins of Western Greece, northeastern Mediterranean Sea

    NASA Astrophysics Data System (ADS)

    Lykousis, V.; Roussakis, G.; Sakellariou, D.

    2009-06-01

    Sediment instabilities are common on the prodeltas of the seismically active continental margins of Western Greece. Sediment failures on the low-angle (0.5°-2°) prodelta slopes manifest themselves as successions of peripheral rotational block slumps restricted to the foresets of the late highstand systems tract (HST). The individual slump blocks are about 80-150 m long and are bounded by growth faults acting as curved slip planes that extend to a mean depth of 10-15 m below seafloor. Shear planes develop in the lower part of muddy and/or gas charged HST foresets. Deeper basal transparent muddy layers of the early HST bottomset, together with the late Pleistocene transgressive systems tract sequences (TST), are mostly unaffected. On the steeper (2°-6°) fan delta slopes of the western Gulf of Corinth debris flows and avalanches with a significant retrogressive component dominate slope destabilisation. Sediment cores taken from landslide scarps and slide planes penetrated gas bubble releasing sediments thereby indicating that failure planes are in the late HST foresets/upper part of the early HST bottomsets gas charged zone. The foresets of the HST prodelta deposits display high water content (30-80%), low bulk density (1.4-1.9 g cm-3) and relatively low values of undrained shear strength (3-20 kPa). The water content of the HST distal muddy bottomsets is relatively higher (50-110%) and bulk density relatively lower (1.3-1.7 g cm-3) with low values of shear strength (2-10 kPa). The shear strength of the gas releasing sediment layer displays lower values (2-9 kPa) relative to the overlying, post failure, muddy sediments of the late 100-300 years. Slope stability was calculated using the normalised soil parameter (NSP) method under undrained conditions for normally consolidated prodelta sediments. This analysis indicates that instabilities could be induced by critical earthquake ground accelerations of 26.6-29.6% g for the HST foresets and 12.4-14.1% g for the basal

  5. 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.

  6. Mechanical constraints on the chronology of fracture activation in folded Devonian sandstone of the western Moroccan Anti-Atlas

    NASA Astrophysics Data System (ADS)

    Guiton, Martin L. E.; Sassi, William; Leroy, Yves M.; Gauthier, Bertrand D. M.

    2003-08-01

    The three-dimensional meter-scale fracture networks, observed on exposed folds between the towns of Tata and Akka, western Moroccan Anti-Atlas, consist mostly of planar discontinuities, which are sub-perpendicular to the bedding and partitioned in three main sets. The chronology of their activation is proposed in five stages since the Hercynian orogeny. Stage 1 predates folding and involves the horizontal compression of the Emsian sandstone. It involves fracture set I, composed of systematic joints parallel to the direction of compression. Stages 2-4 correspond to the folding and are marked in the outer-arc by the activation of fracture set II, composed mainly of joints parallel to the fold axial plane. Stage 5 is a regional shear event during which sets I and III, separated by an angle close to 60°, are activated in a conjugate manner. To throw light on the recurrent difficulty in discriminating between activation of inherited and new fractures, an elasto-plastic model is used to construct a stress path in the pervasively fractured medium idealized as a continuum. Each fracture set obeys the Mohr-Coulomb criterion truncated in tension to describe both sliding and opening activations. Finite-element simulations of a simple buckling event accounting for the field fracture sets are presented. It is shown that set I cannot be generated by folding and thus does belong to stage 1. Set II is activated at a later stage of folding than expected from the field interpretation. Set III cannot be activated during stage 2, confirming its role in stage 5. The advantages and limitations of the proposed modeling are finally discussed.

  7. 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.

  8. Urinary antibiotic activity in paediatric patients attending an outpatient department in north-western Cambodia

    PubMed Central

    Emary, Katherine R W; Carter, Michael J; Pol, Sreymom; Sona, Soeng; Kumar, Varun; Day, Nicholas P J; Parry, Christopher M; Moore, Catrin E

    2015-01-01

    Objective Antibiotic resistance is a prominent public and global health concern. We investigated antibiotic use in children by determining the proportion of unselected children with antibacterial activity in their urine attending a paediatric outpatient department in Siem Reap, Cambodia. Methods Caregiver reports of medication history and presence of possible infection symptoms were collected in addition to urine samples. Urine antibiotic activity was estimated by exposing bacteria to urine specimens, including assessment against multiresistant bacteria previously isolated from patients in the hospital (a methicillin-resistant Staphylococcus aureus (MRSA), a multiresistant Salmonella typhi and an extended-spectrum β-lactamase (ESBL)-producing Escherichia coli isolate). Results Medication information and urine were collected from 775 children. Caregivers reported medication use in 69.0% of children in the preceding 48 h. 31.7% samples showed antibacterial activity; 16.3% showed activity against a local multiresistant organism. No specimens demonstrated activity against an ESBL-producing E. coli. Conclusions Antibiotics are widely used in the community setting in Cambodia. Parents are often ill-informed about drugs given to treat their children. Increasing the regulation and training of private pharmacies in Cambodia may be necessary. Regional surveillance of antibiotic use and resistance is also essential in devising preventive strategies against further development of antibiotic resistance, which would have both local and global consequences. PMID:25324202

  9. 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…

  10. 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.

  11. Novel Bacillus thuringiensis Binary Insecticidal Crystal Proteins Active on Western Corn Rootworm, Diabrotica virgifera virgifera LeConte

    PubMed Central

    Ellis, R. Tracy; Stockhoff, Brian A.; Stamp, Lisa; Schnepf, H. Ernest; Schwab, George E.; Knuth, Mark; Russell, Josh; Cardineau, Guy A.; Narva, Kenneth E.

    2002-01-01

    A new family of insecticidal crystal proteins was discovered by screening sporulated Bacillus thuringiensis cultures for oral activity against western corn rootworm (WCR) larvae. B. thuringiensis isolates PS80JJ1, PS149B1, and PS167H2 have WCR insecticidal activity attributable to parasporal inclusion bodies containing proteins with molecular masses of ca. 14 and 44 kDa. The genes encoding these polypeptides reside in apparent operons, and the 14-kDa protein open reading frame (ORF) precedes the 44-kDa protein ORF. Mutagenesis of either gene in the apparent operons dramatically reduced insecticidal activity of the corresponding recombinant B. thuringiensis strain. Bioassays performed with separately expressed, biochemically purified 14- and 44-kDa polypeptides also demonstrated that both proteins are required for WCR mortality. Sequence comparisons with other known B. thuringiensis insecticidal proteins failed to reveal homology with previously described Cry, Cyt, or Vip proteins. However, there is evidence that the 44-kDa polypeptide and the 41.9- and 51.4-kDa binary dipteran insecticidal proteins from Bacillus sphaericus are evolutionarily related. The 14- and 44-kDa polypeptides from isolates PS80JJ1, PS149B1, and PS167H2 have been designated Cry34Aa1, Cry34Ab1, and Cry34Ac1, respectively, and the 44-kDa polypeptides from these isolates have been designated Cry35Aa1, Cry35Ab1, and Cry35Ac1, respectively. PMID:11872461

  12. Novel Bacillus thuringiensis binary insecticidal crystal proteins active on western corn rootworm, Diabrotica virgifera virgifera LeConte.

    PubMed

    Ellis, R Tracy; Stockhoff, Brian A; Stamp, Lisa; Schnepf, H Ernest; Schwab, George E; Knuth, Mark; Russell, Josh; Cardineau, Guy A; Narva, Kenneth E

    2002-03-01

    A new family of insecticidal crystal proteins was discovered by screening sporulated Bacillus thuringiensis cultures for oral activity against western corn rootworm (WCR) larvae. B. thuringiensis isolates PS80JJ1, PS149B1, and PS167H2 have WCR insecticidal activity attributable to parasporal inclusion bodies containing proteins with molecular masses of ca. 14 and 44 kDa. The genes encoding these polypeptides reside in apparent operons, and the 14-kDa protein open reading frame (ORF) precedes the 44-kDa protein ORF. Mutagenesis of either gene in the apparent operons dramatically reduced insecticidal activity of the corresponding recombinant B. thuringiensis strain. Bioassays performed with separately expressed, biochemically purified 14- and 44-kDa polypeptides also demonstrated that both proteins are required for WCR mortality. Sequence comparisons with other known B. thuringiensis insecticidal proteins failed to reveal homology with previously described Cry, Cyt, or Vip proteins. However, there is evidence that the 44-kDa polypeptide and the 41.9- and 51.4-kDa binary dipteran insecticidal proteins from Bacillus sphaericus are evolutionarily related. The 14- and 44-kDa polypeptides from isolates PS80JJ1, PS149B1, and PS167H2 have been designated Cry34Aa1, Cry34Ab1, and Cry34Ac1, respectively, and the 44-kDa polypeptides from these isolates have been designated Cry35Aa1, Cry35Ab1, and Cry35Ac1, respectively. PMID:11872461

  13. Influence of monsoon over the warm pool on interannual variation on tropical cyclone activity over the western North Pacific

    NASA Astrophysics Data System (ADS)

    Chen, Guanghua; Huang, Ronghui

    2008-03-01

    The relationship between the interannual variation in tropical cyclone (TC) activity over the western North Pacific (WNP) and the thermal state over the warm pool (WP) is examined in this paper. The results show that the subsurface temperature in the WP is well correlated with TC geographical distribution and track type. Their relation is linked by the East Asian monsoon trough. During the warm years, the westward-retreating monsoon trough creates convergence and vorticity fields that are favorable for tropical cyclogenesis in the northwest of the WNP, whereas more TCs concentrating in the southeast result from eastward penetration of the monsoon trough during the cold years. The steering flows at 500 hPa lead to a westward displacement track in the warm years and recurving prevailing track in the cold years. The two types of distinct processes in the monsoon environment triggering tropical cyclogenesis are hypothesized by composites centered for TC genesis location corresponding to two kinds of thermal states of the WP. During the warm years, low-frequency intraseasonal oscillation is active in the west of the WNP such that eastward-propagating westerlies cluster TC genesis in that region. In contrast, during the cold years, the increased cyclogenesis in the southeast of the WNP is mainly associated with tropical depression type disturbances transiting from equatorially trapped mixed Rossby gravity waves. Both of the processes may be fundamental mechanisms for the inherent interannual variation in TC activity over the WNP.

  14. The effects of solar activity on the global solar radiation measured at Khargha Oasis in the Western Dessert of Egypt

    NASA Astrophysics Data System (ADS)

    Shaltout, M.; Mohamed, A.

    Khargha is an Oasis in the Western Desert of Egypt of coordinates lat. 25 o 27/ N, long. 30 o 32 / E, and elevation 77.8 meter over the sea level. It is one of the driest areas in the world, the global solar radiation measured starting from January 1976 till now by station belong to the Egyptian Meteorological Authority. We used the data for the last 25 years of the 20"' Century on the daily bases, it is more than two solar cycles. The annual mean of relative humidity for Khargha is 30, and the total rainfall in mms as annual mean is less than one. Where, the evaporation in mms per day as annual mean is about 16. The total sky cover in oktas as annual mean is 0.4 at the midnight, while it is one oktas at the noon as 2annual mean, and 0.7 oktas on the mean of the day. The annual mean is 6.5 Kwh/rn /day for global solar radiation. Fourier analysis technique used to analysis the time series to show any reflection for the 11-year cycle of the solar activity on the measured global radiation in remote, clean, and dry desert area. The results indicate periodicity's similar to the solar activity periodicities, especially that of the eleven year cycle, in a good indication for the effect of solar activity on the climate change.

  15. L-Asparaginase Activity of Fungal Endophytes from Tabernaemontana heyneana Wall. (Apocynaceae), Endemic to the Western Ghats (India)

    PubMed Central

    Manasa, Chandramouli; Nalini, Monnanda Somaiah

    2014-01-01

    “Endophytes,” the microbes residing within the plant tissues, are important sources of secondary metabolites. Tabernaemontana heyneana Wall., a medicinal tree, endemic to the Western Ghats with rich ethnobotanical history and unique chemical diversity, was selected to study fungal endophytes and evaluate them for L-asparaginase activity. Healthy plant parts were selected for the isolation of endophytes following standard isolation protocols. A total of 727 isolates belonging to 20 taxa were obtained. The isolates comprised of bark (11%), twig (22%), leaf (43%), fruit (12.0%), and seeds (12%). Endophytes such as Colletotrichum, Curvularia, Fusarium, Phomopsis, Verticillium, and Volutella colonized T. heyneana plant parts. Fusarium sp., Phomopsis spp., isolate Thlf01, and Fusarium solani were the dominant genera of bark, twig, leaf, fruits, and seed samples, respectively. The endophytes were screened for their ability to utilize L-asparagine by plate assay method. Fusarium spp. exhibited a high level of activity among the nine endophytes tested positive for L-asparaginase activity. Studies underline the potentials of endophyte-derived fungal L-asparaginases as sources of chemotherapeutic agents. PMID:27382605

  16. 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.

  17. 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

  18. 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

  19. Western-blot detection of PrP**sc in archived paraffin-embedded brainstem from scrapie-affected sheep

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Scrapie is a naturally occurring fatal neurodegenerative disease of adult sheep and goats, one of a group of mammalian diseases known as transmissible spongiform encephalopathies or prion diseases. Immunoassays that identify disease-associated prion protein (PrP**Sc) are integral to the diagnosis o...

  20. 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...

  1. Detection of the disease-associated isoform of the prion protein in formalin-fixed tissues by Western blot

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Clinical signs of prion disease are not pathognomonic and include a variety of differential diagnoses. Specific immune responses have not been detected in affected organisms, serological tests to obtain evidence for the presence of the infectious agent are not available, and nucleic acid-based detec...

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

    PubMed Central

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

    2013-01-01

    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 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 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

  3. Western Mediterranean coastal waters--monitoring PCBs and pesticides accumulation in Mytilus galloprovincialis by active mussel watching: the Mytilos project.

    PubMed

    Scarpato, Alfonso; Romanelli, Giulia; Galgani, Francois; Andral, Bruno; Amici, Marina; Giordano, Pierpaolo; Caixach, Josep; Calvo, Monica; Campillo, Juan Antonio; Albadalejo, José Benedicto; Cento, Alessandro; BenBrahim, Samir; Sammari, Cherif; Deudero, Salud; Boulahdid, Mostefa; Giovanardi, Franco

    2010-04-01

    In order to evaluate the contamination levels in the Western Mediterranean basin, the active mussel watch methodology has been applied. This methodology consists of mussel transplantation (Mytilus galloprovincialis) from non impacted areas to selected coastal areas, characterised by potential impact from the continent due to contaminating sources. The areas of interest were selected along the entire coastal development of the Western Mediterranean sea, 122 sites in total. The time of mussel caging exposure was 12 weeks. The project was co-financed in the frame of the Interreg IIIB Meddoc Programme, aimed at determining the overall chemical quality of the Mediterranean sea, consistent with the Water Framework Directive 2000/60. Several partners representative of the coastal Mediterranean Countries were involved in the Project, with the purpose of building up a common surveillance network, adopting shared methodologies. In this paper we present the results of three yearly monitoring campaigns (2004, 2005, 2006) carried out along the coasts of Italy, France, Spain, Morocco, Algeria and Tunisia, including the coastal environment of Baleares, Sicily, Sardinia and Corsica. The contamination levels of Pesticides (DDT and its metabolites, Hexachlorocyclohexane isomers alpha and gamma) and Polychlorinated biphenyls, are reported and discussed. Statistical elaborations performed on the original data set were mainly aimed at validating the raw sample distributions, by means of the Johnson method. Both DD and PCB species frequency distributions have been approximated to appropriate theoretical distributions, belonging to the Log-normal and Bounded families. By integrating the related Probability Density Functions (p.d.f.), different accumulation values for DDT, DDD and DDE and PCB species have been estimated, corresponding to fixed percentage points of the area under the respective curves. By choosing appropriate probability level boundaries (33rd and 66th percentile

  4. 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

  5. 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

  6. 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

  7. 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.

  8. Biologically active warm-core anticyclonic eddies in the marginal seas of the western Pacific Ocean

    NASA Astrophysics Data System (ADS)

    Chen, Yuh-ling Lee; Chen, Houng-Yung; Jan, Sen; Lin, Yen-Huei; Kuo, Tien-Hsia; Hung, Jia-Jang

    2015-12-01

    Our investigations in the northern South China Sea (SCS) have revealed warm-core anticyclonic eddies that had a depressed pycnocline and a high biological productivity and phytoplankton abundance. With an elliptical shape of 420-430 km in major axis and 240-260 km in minor axis, these eddies were formed in the winter as the Kuroshio Current intruded through the Luzon Strait into the SCS under the prevailing northeast monsoon. They were characterized by a deep mixed layer up to 140-180 m, in which nitrate was relatively abundant. Although chlorophyll a concentration per volume of seawater was not always higher inside than outside the eddies, water-column (0-200 m) integrated chlorophyll a concentration and abundances of Synechococcus, coccolithophores, and diatoms were higher inside than outside the eddies. Primary productivity and nitrate-uptake new production inside the eddies were higher than or equal to those outside the eddies. Unlike the mode-water anticyclonic eddy that is biologically productive with a domed shallow seasonal pycnocline, the eddies we investigated had high surface temperatures and depressed pycnoclines in the upper water column. Possible explanations for these biological aspects were that the eddies were at their decaying stage, the eddies re-incorporated intermittently with an intruding Kuroshio branch, or the passage of the prevalent high amplitude internal tides introduced nutrients to the eddies. Frequent occurrences of eddies in oceanic regimes, especially cold eddies, are associated with high biological activity. Some warm eddies, such as these investigated in the present study, also have high biological activities, indicating that more rigorous in situ studies relating to eddy biological activity are needed in ocean regimes such as the SCS, where a half of the eddies are warm eddies.

  9. 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.

  10. 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.

  11. 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. PMID:15294828

  12. Binary Toxins from Bacillus thuringiensis Active against the Western Corn Rootworm, Diabrotica virgifera virgifera LeConte

    PubMed Central

    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

    2004-01-01

    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. PMID:15294828

  13. 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. PMID:21563213

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

    PubMed

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

    2015-01-01

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

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

    SciTech Connect

    Gray, M.R. )

    1992-02-01

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

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

    EPA Science Inventory

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

  17. 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

  18. Morphology of turbidite systems within an active continental margin (the Palomares Margin, western Mediterranean)

    NASA Astrophysics Data System (ADS)

    Perez-Hernandez, S.; Comas, M. C.; Escutia, C.

    2014-08-01

    The Palomares Margin, an NNE-SSW segment of the South Iberian Margin located between the Alboran and the Algerian-Balearic basins, is dissected by two major submarine canyon systems: the Gata (in the South) and the Alías-Almanzora (in the North). New swath bathymetry, side-scan sonar images, accompanied by 5 kHz and TOPAS subbottom profiles, allow us to recognize these canyons as Mediterranean examples of medium-sized turbidite systems developed in a tectonically active margin. The Gata Turbidite System is confined between residual basement seamounts and exhibits incised braided channels that feed a discrete deep-sea fan, which points to a dominantly coarse-grained turbiditic system. The Alías-Almanzora Turbidite System, larger and less confined, is a good example of nested turbiditic system within the canyon. Concentric sediment waves characterize the Alías-Almanzora deep-sea fan, and the size and acoustic character of these bedforms suggest a fine-grained turbidite system. Both canyons are deeply entrenched on a narrow continental shelf and terminate at the base of the continental slope as channels that feed deep sea fans. While the Alías-Almanzora Turbidite System is the offshore continuation of seasonal rivers, the Gata Turbidite System is exclusively formed by headward erosion along the continental slope. In both cases, left-lateral transpressive deformation influences their location, longitudinal profiles, incision at the upper sections, and canyon bending associated with specific fault segments.

  19. 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.

  20. 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

  1. Antimicrobial properties, antioxidant activity and bioactive compounds from six wild edible mushrooms of western ghats of Karnataka, India

    PubMed Central

    Ramesh, Ch.; Pattar, Manohar G.

    2010-01-01

    Methanolic extracts of 6 wild edible mushrooms isolated from the Western Ghats of Karnataka were used in this study. Among the isolates (Lycoperdon perlatum, Cantharellus cibarius, Clavaria vermiculris, Ramaria formosa, Marasmius oreades, Pleurotus pulmonarius), only 4 showed satisfactory results. Quantitative analysis of bioactive components revealed that total phenols are the major bioactive component found in extracts of isolates expressed as mg of GAE per gram of fruit body, which ranged from 3.20 ± 0.05 mg/mL to 6.25 ± 0.08 mg/mL. Average concentration of flavonoid ranged from 0.40 ± 0.052 mg/mL to 2.54 ± 0.08 mg/mL; followed by very small concentration of ascorbic acid (range, 0.06 ± 0.01 mg/mL to 0.16 ± 0.01 mg/mL) in all the isolates. All the isolates showed high phenol and flavonoid content, but ascorbic acid content was found in traces. Antioxidant efficiency by inhibitory concentration on 1,1-Diphenly-2-picrylhydrazyl (DPPH) was found significant when compared to standard antioxidant like Buthylated hydroxyanisol (BHA). The concentration (IC50) ranged from 0.94 ± 0.27 mg/mL to 7.57 ± 0.21 mg/mL. Determination of antimicrobial activity profile of all the isolates tested against a panel of standard pathogenic bacteria and fungi indicated that the concentrations of bioactive components directly influence the antimicrobial capability of the isolates. Agar diffusion assay showed considerable activity against all bacteria. Minimum inhibitory concentration values of the extracts of 4 isolates showed that they are also active even in least concentrations. These results are discussed in relation to therapeutic value of the studied mushrooms. PMID:21808550

  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 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

  4. Evolution of the Late Pleistocene Aspe River (Western Pyrenees, France). Signature of climatic events and active tectonics

    NASA Astrophysics Data System (ADS)

    Nivière, Bertrand; Lacan, Pierre; Regard, Vincent; Delmas, Magali; Calvet, Marc; Huyghe, Damien; Roddaz, Bernard

    2016-03-01

    We make use of the cosmogenic nuclide 10Be exposure to date an alluvial terrace of the Aspe River in the foothills of the northwestern Pyrenees. Initially ascribed to the Rissian glaciation, our dating shows that the terrace was abandoned at 18 ± 2 kyr. In reference to the Late Pleistocene climatic chronology, two kinds of terraces can be distinguished: high-standing fill terraces probably deposited during glacial events and lower cut-in-fill and strath terraces cut during the postglacial river incision. A part of the terrace aggradations could have occurred during the Würmian glacial episodes. Hence, the dated terrace fits in with the prevailing view of incision during climate transitions. Our study also shows that elevation is not a good criterion of terrace correlation, which should be better carried out on the basis of absolute dating. In addition, this dating also suggests a potential Late Pleistocene fault reactivation of the Mail Arrouy thrust in this tectonically active area of the Western Pyrenees.

  5. 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

  6. 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).

  7. 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.

  8. 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. PMID:26756993

  9. Dioxins, furans and AHH-active PCB congeners in eggs of two gull species from the western Mediterranean.

    PubMed

    Pastor, D; Ruiz, X; Barceló, D; Albaigés, J

    1995-09-01

    Polychlorinated biphenyls, dibenzodioxins and dibenzofurans were analysed in eggs of a protected gull species, the Audouin's Gull (Larus audouinii) and compared to those of the Yellow-legged Gull (Larus cachinnans), both breeding in the Western Mediterranean (Ebro Delta and Medes Islands, respectively). Differences in concentrations as well as in congener profiles reflected differences in both habitat and diet of the two species. Levels of AHH-active PCB congeners were lower in Yellow-legged Gull (0.4-1.6 micrograms/g d.w) than in Audouin's Gull eggs (1.2-33.9 micrograms/g d.w.). These concentrations, expressed in international toxic equivalence factors (i-TEQ/g d.w.), were on average 24 times higher in the Audouin's gull. I-TEQ levels due to dioxins were also higher in this species by a factor of ca. 7. I-TEQ levels related to PCBs resulted 90-230 times higher than those of dioxins and furans. Thus, AHH-inducing PCBs might represent even higher toxicological hazards than dioxins and furans to some populations of seabirds. The necessity of assessing the impact of these compounds in rare and protected species is pointed out. PMID:8528648

  10. 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

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

    PubMed

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

    2016-01-01

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

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

    PubMed Central

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

    2016-01-01

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

  13. 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

  14. 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.

  15. Analysis of multitemporal laserscanned DTMs of an active landslide (Doren, Western Austria) using a robust planefitting segmentation

    NASA Astrophysics Data System (ADS)

    Koma, Zs.; Pocsai, A.; Székely, B.; Dorninger, P.; Zámolyi, A.; Roncat, A.

    2012-04-01

    Structural geomorphometric analysis of high-resolution laser scanned DTMs is a straightforward method to study microtopographic components of dynamically forming landscapes, and thus areas affected by mass movements. However, results for multitemporal DTMs may turn out to be difficult to evaluate. In our approach, a robust plane fitting algorithm is used to create various segmentations to filtered lidar point cloud (ground surface points) by applying different sets of parameters. The resulting sets of planes are analyzed in terms of their geologic meaning and compared in order to detect changes. Our study area, the Doren landslide (Bregenzerwald, Vorarlberg, Western Austria), an actively forming landslide developed in molasse sediments has been measured several times by laser scanning (lidar). These DTMs form the input to our procedure. The DTMs are analyzed by the segmentation algorithm, using varying parameter sets (i.e. number of minimum points, standard deviation, point-to-plain distance). The segmented results are checked for indications of geological structures as well as for features belonging to the moving material of the landslide. Finally the segments of the different years are compared. Results show that patterns composed of segments of steep and less steep valley sides can be correlated with the tectonic and lithological setting of the study area. Furthermore some narrow linear or curvilinear zones appear that can be related to the outlines of some small internal mass movements. Interestingly, the various years show sometimes similar patterns despite the continuous displacement of the sliding material. The project has been supported by the Austrian Academy of Sciences (ÖAW) in the framework of the project "Geophysik der Erdkruste".

  16. An active extensional deformation example: 19 May 2011 Simav earthquake (Mw = 5.8), Western Anatolia, Turkey

    NASA Astrophysics Data System (ADS)

    Demirci, Alper; Özden, Süha; Bekler, Tolga; Kalafat, Doğan; Pınar, Ali

    2015-08-01

    The Simav Earthquake that occurred on 19 May 2011 in western Turkey was investigated on the basis of seismological data and geological observations. Approximately WNW-ESE trending surface ruptures were observed on the Simav Fault. The focal mechanism parameters of the earthquake (Mw = 5.8) and its aftershocks (Mw  >  3.5) were estimated using time-domain moment tensor inversion. A total of 2245 events were located with Geiger’s conventional absolute location method then relocated using the double difference (DD) algorithm. The calculated locations at a depths between 2 and 16 km were found to be consistent with Coulomb stress variation in the area. Average variance reduction (VR) of the solutions was calculated as ~70%. The focal parameters of strike dip and slip of the main shock, occurring at a depth of 11 km dipping towards the NNE, were estimated at 277, 62 and -92, respectively. The most striking indication of the study is that the area is dominated by normal faults with mainly WNW-ESE trends. It is also concluded that earthquakes in the region are caused by an active and regional NNE-SSW (N 12° E) trending (σ3 axis) extension regime. The mean stress ratio is 0.80, indicating a triaxial stress state. This extension is probably associated with a slab-pull force and /or roll-back due to the complex subduction process of the African Plate beneath Anatolian block along both the Hellenic and Cyprus arcs in the eastern Mediterranean region.

  17. 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.

  18. 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.

  19. 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

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

    SciTech Connect

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

    1993-07-01

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

  1. 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

  2. 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

  3. EFFECTS OF AMMONIUM AND NITRATE ON NUTRIENT UPTAKE AND ACTIVITY OF NITROGEN ASSIMILATING ENZYMES IN WESTERN HEMLOCK

    EPA Science Inventory

    Western hemlock seedlings were grown in nutrient solutions with ammonium, nitrate or ammonium plus nitrate as nitrogen sources. he objectives were to examine (1) possible selectivity for ammonium or nitrate as an N source, (2) the maintenance of charge balance during ammonium and...

  4. Reproduction of the Western Tarnished Plant Bug, Lygus hesperus, in Relation to Age, Gonadal Activity and Mating Status.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Understanding the basic life history and underlying regulatory mechanisms for a pest insect is essential for developing targeted control strategies, but for many insects relatively little is known. Although the western tarnished plant bug, Lygus hesperus Knight (Heteroptera: Miridae) has a substant...

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

    SciTech Connect

    van den Berg, K.J.

    1986-05-15

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

  6. 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

  7. Estimation of active faulting in a slow deformation area: Culoz fault as a case study (Jura-Western Alps junction).

    NASA Astrophysics Data System (ADS)

    de La Taille, Camille; Jouanne, Francois; Crouzet, Christian; Jomard, Hervé; Beck, Christian; de Rycker, Koen; van Daele, Maarten; Lebourg, Thomas

    2014-05-01

    The north-western Alps foreland is considered as still experiencing distal effects of Alpine collision, resulting in both horizontal and vertical relative displacements. Based on seismological and geodetic surveys, detailed patterns of active faulting (including subsurface décollements, blind ramps and deeper crustal thrusts have been proposed (Thouvenot et al., 1998), underlining the importance of NW-SE left-lateral strike-slip offsets as along the Vuache and Culoz faults (cf. the 1996 Epagny event: M=5.4; Thouvenot et al., 1998 and the 1822 Culoz event I=VII-VIII; Vogt, 1979). In parallel to this tectonic evolution, the last glaciation-deglaciation cycles contributed to develop large and over-deepened lacustrine basins, such as Lake Le Bourget (Perrier, 1980). The fine grain, post LGM (ie post 18 ky), sedimentary infill gives a good opportunity to evidence late quaternary tectonic deformations. This study focuses on the Culoz fault, extending from the Jura to the West, to the Chautagne swamp and through the Lake Le Bourget to the East. Historical earthquakes are known nearby this fault as ie the 1822 Culoz event. The precise location and geometry of the main fault is illustrated but its Eastern termination still needs to be determined. High resolution seismic sections and side-scan sonar images performed in the 90's (Chapron et al., 1996) showed that the Col du Chat and Culoz faults have locally deformed the quaternary sedimentary infill of the lake. These studies, mainly devoted to paleo-climate analysis were not able to determine neither the geometry of the fault, or to quantify the observed deformations. A new campaign devoted to highlight the fault geometry and associated deformation, has been performed in October 2013. Very tight profiles were performed during this high resolution seismic survey using seistec boomer and sparker sources. In several places the rupture reaches the most recent seismic reflectors underlying that these faults were active during

  8. 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

    2015-11-01

    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.

  9. 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.

  10. 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

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

    SciTech Connect

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

    1990-01-01

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

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

    PubMed Central

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

    1999-01-01

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

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

    SciTech Connect

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

    1986-10-01

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

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

    SciTech Connect

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

    1994-09-01

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

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

    PubMed

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

    2007-01-01

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

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

    PubMed

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

    2009-11-01

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

  17. Western USA

    Atmospheric Science Data Center

    2014-05-15

    article title:  Western United States Beyond the Four Corners ... to examine Earth's land, oceans, atmosphere, ice, and life as a total integrated system. MISR was built and is managed by NASA's ... D.C. The Terra spacecraft is managed by NASA's Goddard Space Flight Center, Greenbelt, MD. The MISR data were obtained from the NASA ...

  18. 1.2 million years of aeolian activity in northwestern Western Australian recorded in a deep-sea core in the eastern Indian Ocean

    NASA Astrophysics Data System (ADS)

    Temmesfeld, Felix; Stuut, Jan-Berend; Bassinot, Franck; Heslop, David; De Deckker, Patrick

    2015-04-01

    There is no continuous record of aridity from the Australian continent and we are presenting here, for the first time, a record of aeolian activity for northwestern Western Australia. Our data are based on a 32m long deep-sea core taken offshore North West Cape, the northwestern tip of Western Australia. Our data rely on 2 adjacent studies: (a) a complete XRF scan of the core which provide elemental ratios which are translated into an aeolian component as well as a fluvial discharge-to-sea component. The first one relates to periods of aridity inland Australia, whereas the second one is interpreted as river discharge during periods of monsoonal activity; (b) a close examination of grain size analyses and observations of wind-blown quartz grains found in close to 600 samples taken from the core. Our results clearly show a cyclic record of alternating dry and wet periods spanning the last 1.2 million years. Our findings also indicate that monsoonal activity as well as desertification were already in place in northern Australia so long ago, and this has clear implications for the evolution of the arid zone biota, associated fire activities and geomorphological features in northern Australia.

  19. Western Analysis of Histone Modifications (Aspergillus nidulans)

    PubMed Central

    Soukup, Alexandra; Keller, Nancy P.

    2016-01-01

    Western blotting allows for the specific detection of proteins and/or modifications of proteins by an antibody of interest. This protocol utilizes a crude nuclei extraction protocol for Aspergillus nidulans to enrich for histones and other nuclear proteins prior to gel electrophoresis. Post translational modifications of histones may then be easily detected. After electrophoresis, the selected antibodies are used to detect and quantify levels of the modifications of interest.

  20. 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

  1. 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

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

    SciTech Connect

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

    1994-09-01

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

  3. 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

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

    PubMed Central

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

    2000-01-01

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

  5. 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.

  6. 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.

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

    PubMed

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

    2011-06-15

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

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

    SciTech Connect

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

    1984-03-01

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

  9. 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.

  10. 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

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

    PubMed Central

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

    1992-01-01

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

  12. The effect of anthropogenic activity on the occurrence of Culicoides species in the South-Western Khomas Region, Namibia.

    PubMed

    Becker, Elbè; Venter, Gert J; Labuschagne, Karien; Greyling, Telanie; van Hamburg, Huib

    2013-01-01

    Certain species of midges in the genus Culicoides (Diptera: Ceratopogonidae) are vectors of several serious orbiviral (Reoviridae) diseases, one of which, African horse sickness (AHS), was reported in the South-Western area of Khomas Region, Namibia, where it had been believed to be absent. Culicoides imicola, AHS principal vector, was collected in several farms in the area during the winter of 2009. The objective of this study was to determine whether Culicoides midges, especially C. imicola, were favoured at anthropogenic impacted/homestead sites in the arid Khomas Region, where they were not expected to occur under natural, veld conditions. The natural 'background' Culicoides communities where determined from collections made at veld sites, which were then compared to corresponding collections made at homestead sites. Altogether, 10,178 Culicoides midges were collected at homesteads and were then compared to 1,733 individuals collected at veld sites. Culicoides midge numbers were likely boosted in anthropogenic impacted areas/homesteads. This was also the case for the Culicoides species that are vector of AHS. This study indicated the significance of human settlement in the Khomas Region in terms of Culicoides midge abundance and distribution and showed the implications that this may have on the transmission of Culicoides-vectored diseases. PMID:24166479

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

    PubMed Central

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

    2006-01-01

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

  14. 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

  15. [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

  16. 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.

  17. 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.

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

    PubMed

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

    1995-01-01

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

  19. 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...

  20. 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

  1. 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

    2015-09-01

    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

  2. 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 Central

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

    2015-01-01

    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

  3. 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

  4. 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

  5. Insecticidal and repellent activities of pyrethroids to the three major pyrethroid-resistant malaria vectors in western Kenya

    PubMed Central

    2014-01-01

    Background The dramatic success of insecticide treated nets (ITNs) and long-lasting insecticidal nets (LLINs) in African countries has been countered by the rapid development of pyrethroid resistance in vector mosquitoes over the past decade. One advantage of the use of pyrethroids in ITNs is their excito-repellency. Use of the excito-repellency of pyrethroids might be biorational, since such repellency will not induce or delay the development of any physiological resistance. However, little is known about the relationship between the mode of insecticide resistance and excito-repellency in pyrethroid-resistant mosquitoes. Methods Differences in the reactions of 3 major malaria vectors in western Kenya to pyrethroids were compared in laboratory tests. Adult susceptibility tests were performed using World Health Organization (WHO) test tube kits for F1 progenies of field-collected An. gambiae s.s., An. arabiensis, and An. funestus s.s., and laboratory colonies of An. gambiae s.s. and An. arabiensis. The contact repellency to pyrethroids or permethrin-impregnated LLINs (Olyset® Nets) was evaluated with a simple choice test modified by WHO test tubes and with the test modified by the WHO cone bioassay test. Results Field-collected An. gambiae s.s., An. arabiensis, and An. funestus s.s. showed high resistance to both permethrin and deltamethrin. The allelic frequency of the point mutation in the voltage-gated sodium channel (L1014S) in An. gambiae s.s. was 99.3–100%, while no point mutations were detected in the other 2 species. The frequency of takeoffs from the pyrethroid-treated surface and the flying times without contacting the surface increased significantly in pyrethroid-susceptible An. gambiae s.s. and An. arabiensis colonies and wild An. arabiensis and An. funestus s.s. colonies, while there was no significant increase in the frequency of takeoffs or flying time in the An. gambiae s.s. wild colony. Conclusion A different repellent reaction was observed in

  6. Sensing vegetation growth and senescence with reflected GPS signals: Active microwave detection of western North America phenology

    NASA Astrophysics Data System (ADS)

    Evans, Sarah Grace

    We explore a new technique to estimate vegetation growth and senescence using reflected GPS signals (multipath) measured by geodetic-quality GPS stations. The operational GPS-IR statistic Normalized Microwave Reflection Index (NMRI), a measure of multipath scattering, exhibits a clear seasonal cycle as is expected for vegetation growth and senescence. The sensing footprint is ˜1000 m 2, larger than that provided by typical in situ observations but smaller than that from space-based products. Since GPS satellites transmit L-band signals, the vegetation estimates derived from GPS reflections provide global phenology monitoring that is sensitive to changes in vegetation canopy water content and biomass. However, GPS reflections are insensitive to plant greenness, clouds, atmosphere, and solar illumination constraints that adversely affect optical-infrared remote sensing vegetation indices like Normalized Difference Vegetation Index (NDVI). Temporal and spatial diffuse scattering of microwave GPS-IR index NMRI and MODIS-based NDVI is documented at both the site-by-site and regional scale at 184 sites over the western United States. We derive NMRI and NDVI range, correlation between NMRI and NDVI signals, and phenology parameters including: start of season, season length, and peak day of year of vegetation growth. These phenology indexes are compared over a five water-year time series (2008 to 2012) to gauge spatial and temporal offsets. Average correlations (R 2=0.527) were found with NMRI variations lagging NDVI by approximately 21 days. This is consistent with the idea that greenup precedes plant growth. Phenology metrics extracted by microwave NMRI record a later start of season, later peak day of year, and shorter season length than determined by optical NDVI. Metrics are offset spatially with the largest offsets along Pacific Ocean coastline, decreasing inland and subdivided by region, supporting that plant growth cycles are controlled by regional climates. This

  7. Evaluation of antioxidant activity, phenolic, mineral contents and some physicochemical properties of several pine honeys collected from Western Anatolia.

    PubMed

    Akbulut, Mehmet; Ozcan, Mehmet Musa; Coklar, Hacer

    2009-11-01

    In this study, the qualities of 15 red pine honey samples from different parts of the Muğla province in Turkey were evaluated. Important chemical parameters such as degrees Brix, pH, moisture, ash, electrical conductivity, free acidity, proteins, diastase activity, hydroxymethylfurfurale, total reducing sugar, reducing sugar, sucrose, the five common physical parameters refractive index, density, water activity, dynamic viscosity and color index (L*, a* and b*), and mineral compositions, including potassium, sodium, magnesium, phosphorus, calcium, iron, copper, zinc, manganese, nickel, aluminum, boron, molybdenum, and chromium, were determined. Pine honey samples also were analyzed for total phenolic contents and antioxidant activity. The antioxidant activities were evaluated based on the ability of the pine honey extracts to scavenge 1,1-diphenyl-2-picrylhydrazyl. Significant correlations were obtained between the antioxidant activity and phenolic content (r = 0.887). All samples indicated proper maturity, considering the high soluble solids. The values for ash and electrical conductivity were high (0.24-0.60% and 878-1463 muS/cm, respectively). The concentration of 14 minerals of red pine honeys were determined by inductively coupled plasma-atomic emission spectrometry. All samples contained high amounts of potassium, calcium, phosphorus, sodium, iron, magnesium, aluminum, nickel, and zinc. Within the mineral contents, potassium was quantitatively the most important mineral, having an average content of 3,802 ppm, and accounted for 45% the minerals. PMID:19817637

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

    PubMed Central

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

    2004-01-01

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

  9. Intensity of recreational physical activity in different life periods in relation to breast cancer among women in the region of Western Pomerania

    PubMed Central

    2013-01-01

    Background Recreational physical activity has been consistently associated with lower breast cancer risk, but there is a need to study the intensity and duration of activity that are critical to reduce the risk. The aim of this study was to examine the influence of moderate and vigorous intensity of recreational physical activity performed at different age periods on breast cancer risk. Material and methods The case-control study included 858 women with histological confirmation of invasive breast cancer and 1085 women free of any cancer diagnosis, residents of the region of Western Pomerania, aged 28–79 years. The frequency, duration and intensity of lifetime household, occupational and recreational physical activity, sociodemographic characteristics, reproductive factors, family history of breast cancer, current weight and height, and lifestyle habits were measured using a self-administered questionnaire. Unconditional logistic regression analyses were applied to estimate odds ratios (ORs) and 95% confidence intervals (CIs). The risk estimates were controlled for potential risk factors and lifetime household and occupational activities. Results We found a risk reduction for recreational activity done early in life (age periods 14–20, 21–34, 35–50 years), particularly at ages 14–20 and 21–34 years, regardless of intensity. Active women engaging in more than 4.5 hours per week of moderate activity during ages 14–20 years had, on average, a 36% lower risk (OR = 0.64, 95% CI: 0.45–0.89) than women who were never or rarely active. For the women who reported 4.5 hours per week of vigorous activity during this period we found about 64% risk reduction (OR = 0.36, 95% CI: 0.26–0.51). For the period after 50 years of age, recreational activity of moderate or vigorous intensity was not significantly associated with the risk. Conclusions Recreational physical activity of moderate or vigorous intensity done during adolescence, early and middle adulthood

  10. 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

  11. Cyclicity of forest fire occurrence at Kola Peninsula (North-Western Russia) in connection to meteorological and solar activity

    NASA Astrophysics Data System (ADS)

    Shumilov, O. I.; Kasatkina, E. A.; Knyazev, N. V.; Lukina, N. V.

    2010-05-01

    The cyclicity of forest fire number for the period 1958-2007 at Kola Peninsula was investigated. We used the data of regular aerial surveying. The frequency of forest fires was compared with regional meteorological and dendrochronological records. Spectral analysis with help of MEM and wavelet revealed a clear cyclic character of fire occurrence with two main maxima. The main one occurred at frequencies around 18-20 years and the other in the band 2.8-4 year. Detailed analysis showed that fire occurrence at Kola Peninsula was a result of a complicated mixture of both anthropogenic and climatic forcings (temperature and precipitation). Climatic forcing is influenced by variations of solar activity (solar radiation, cosmic rays, cosmic dust etc.). Two maxima in the fire occurrence spectrum seem to be connected to one of the main cycles of solar activity (22 y) and NAO oscillation (3-4 y). As it is well known the NAO variations are rather tightly connected to cyclonic activity in the North Atlantic region. The enhanced numbers of fires were observed close to minima of solar activity. These results may be applied for fire forecasting at Kola Peninsula. This work is financially supported by the Russian Foundation for Basic Research (grant No. 09-04-98801), by the Program of the Russian Academy and by the Regional Scientific Program of Murmansk region.

  12. 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

  13. The use of caves as observatories for recent geodynamic activity and radon gas concentrations in the Western Carpathians and Bohemian Massif.

    PubMed

    Briestensky, M; Thinova, L; Stemberk, J; Rowberry, M D

    2011-05-01

    In recent years, many underground spaces such as caves and deep mines have been used to monitor geodynamic activity in the Western Carpathians and Bohemian Massif. In addition, long-term radon gas monitoring has also being undertaken in three of the caves. The observed radon concentrations have shown diurnal, seasonal and yearly variations. A significant correlation with external temperature has been registered. This pattern is considered to result from the movement of air caused by the contrast between the broadly constant internal and fluctuating external temperatures. The same seasonal effects have been observed in the record of active fault displacements at sites close to the surface. These seasonal effects lead to peak-to-peak massif dilation amplitude that affects the overall trend of fault displacement. It is also noted that a significant decrease in this amplitude occurs with increasing depth beneath the surface. However, high variability has also been observed in both the radon concentration and fault displacement activity recorded. Many events have been registered that link these two geofactors. PMID:21478172

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

    PubMed

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

    2015-10-01

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

  15. 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

  16. 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

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

    PubMed Central

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

    1992-01-01

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

  18. 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

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

    PubMed

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

    1997-08-01

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

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

    PubMed

    Sackstein, Robert; Fuhlbrigge, Robert

    2006-01-01

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

  1. A multidisciplinary study in the geodynamic active western Eger rift (Central Europe): The Quaternary volcanic complex Mytina and the recent CO2-degassing zone Hartousov

    NASA Astrophysics Data System (ADS)

    Flechsig, C.; Heinicke, J.; Kaempf, H. W.; Nickschick, T.; Mrlina, J.

    2013-12-01

    The Eger rift (Central Europe) belongs to the European Cenozoic rift system and represents an approximately 50 km wide and 300 km long ENE-WSW striking continental rift that formed during the Upper Cretaceous-Tertiary transition. This rift zone is one of the most active seismic regions in Central Europe. Especially, the western part of the Eger rift area is dominated by ongoing hidden magmatic processes in the intra-continental lithospheric mantle. Besides of known quaternary volcanoes, these processes take place in absence of any presently active volcanism at the surface. However, they are expressed by a series of phenomena distributed over a relatively large area, like occurrence of repeated earthquake swarms, surface exhalation of mantle-derived and CO2-enriched fluids at mofettes and mineral springs, and enhanced heat flow. At present this is the only known intra-continental region where such deep-seated, active lithospheric processes currently occur. The aim of the project is to investigate the tectonic/geologic near surface structure and the degassing processes of the mofette field of Hartousov, where soil gas measurements (concentration and flux rate) in an area of appr. 3x2 km traced a permeable NS extended segment of a fault zone and revealed highly permeable Diffuse Degassing Structures (DDS). The second target is volcanic environment of the Quaternary volcanic complex Mytina maar and the cinder cone Zelezna hurka/Eisenbühl. The investigations are intended to clarify: a) the spatio-temporal reconstruction of the maar complex, and the palaeo volcanic scenario (geological model, tectonic settings, distribution of pyroclastica, b) the geological structure and the tectonic control of the recent degassing zone, and c) the comperative interpretation of both regions in the consideration of potential future volcanic risk assessment in sub-regions of the western Eger Rift. To investigate both regions the following methods are used: geoelectrics, geomagnetics

  2. 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

  3. Distribution of PTPN22 polymorphisms in SLE from western Mexico: correlation with mRNA expression and disease activity.

    PubMed

    Machado-Contreras, Jesús René; Muñoz-Valle, José Francisco; Cruz, Alvaro; Salazar-Camarena, Diana Celeste; Marín-Rosales, Miguel; Palafox-Sánchez, Claudia Azucena

    2016-08-01

    Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune inflammatory disease characterized by loss of self-tolerance with hyperactivation of autoreactive T and B cells. Protein tyrosine phosphatase non-receptor type 22 (PTPN22) encodes for lymphoid-specific phosphatase (Lyp), which is a key negative regulator of T lymphocyte activation. The aim of this study was to evaluate the genetic contribution of PTPN22 -1123G>C and +1858C>T polymorphisms and their haplotypes in SLE patients, as well as mRNA expression according to -1123G>C promoter polymorphism and disease activity. One hundred and fifty SLE patients and 150 unrelated healthy controls (HC), both Mexican mestizos, were genotyped by PCR-RFLP technique for the PTPN22 -1123G>C and +1858C>T polymorphisms. PTPN22 mRNA expression levels were determined by real-time PCR from PBMCs of thirty patients with SLE and fifteen HC carrying different genotypes. Distributions of genotype and allelic frequencies were similar between SLE and HC. The most frequent alleles were -1123 G and +1858 C in both groups (69 vs. 66 % and 97 vs. 98 %, in SLE and HC, respectively). However, the recessive model of inheritance analysis showed a lower frequency of -1123 CC genotype in SLE patients (7 vs. 15 %), suggesting a protection effect to develop SLE (OR 0.41, CI 1.10-5.28, p = 0.02). Haplotype analysis showed strong linkage disequilibrium D' = 0.98 for PTPN22 -1123G>C and +1858C>T polymorphisms, but haplotypes were not associated with SLE. The PTPN22 mRNA expression did not show differences among -1123G>C genotypes; nevertheless, a significant negative correlation with disease activity was found (r = -0.64, p < 0.01). SLE inactive patients showed similar PTPN22 mRNA expression levels to healthy controls, whereas in patients with severe flare, the expression was nearly depleted. In conclusion, we found a lack of association of PTPN22 -1123G>C and +1858C>T polymorphisms with the risk of developing SLE in a Mexican

  4. 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.

  5. 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.

  6. Microseismic activity analysis for the study of the rupture mechanisms in unstable rock masses (Matterhorn, North-western Alps)

    NASA Astrophysics Data System (ADS)

    Amitrano, D.; Arattano, M.; Chiarle, M.; Mortara, G.; Occhiena, C.; Pirulli, M.; Scavia, C.

    2009-04-01

    Rockfalls are very frequent events in alpine areas and can endanger human lifes and activities. Since high mountains have been affected by an increasing number of these phenomena in the last years, a possible correlation with the effects of climate changes can be hypothesized. The permafrost degradation, causing the thaw of the ice that fills the rock discontinuities, is then investigated among possible causes of rockfalls. Therefore the monitoring of potential rock instabilities in high mountain in relation with permafrost degradation has been carried out in the frame of the Interreg IIIA ALCOTRA "PERMAdataROC" project. Within the project, a monitoring network has been installed in 2007 on the Italian side of the Matterhorn peak, close to the J. A. Carrel refuge (3829 m a.s.l.). This site is an important destination for climbers going to the Matterhorn peak and is frequently affected by rockfall events. The monitoring network consists of a set of 5 triaxial geophones, to record the existing microseismic activity, and one thermometer, to analyze the temperature trend. A preliminary data processing has concerned the classification of the recorded signals, the identification of the most important microseismic events and the analysis of their distribution in time. As far as this last aspect is concerned, first interpretations have evidenced a possible correlation between the temperature trend and the event concentrations, during particular thermal sequences. The research is still in progress and it is expected that a longer recording period of seismic events and temperatures will help to understand if the microseismic activity is mainly concentrated in some periods of the year, in some parts of the slope and if it is produced by superficial or deep events. A concentration of superficial events in some parts of the slope, together with a structural analysis of those portions, could help to focus on the areas that can be more unstable. While, their correlation with

  7. Ion activity products of iron sulfides in groundwaters: Implications from the Choshui fan-delta, Western Taiwan

    NASA Astrophysics Data System (ADS)

    Chen, Wen-Fu; Liu, Tsung-Kwei

    2005-07-01

    Precipitation of iron sulfides is an important process in groundwater geochemistry because it reduces iron mobility in anaerobic aquifers. Iron sulfides occur in various allotropic forms such as amorphous FeS and pyrite, and their solubility products differ up to 13 orders of magnitude. However, few data for ion activity products (IAP) of iron sulfides defined by the equation: H + + FeS (S) = Fe 2+ + HS - in groundwater have been reported in the literature. We computed IAP values of iron sulfides for 46 groundwater samples from the Choshui fan-delta of Taiwan and 65 samples from other areas of the world. The mean of -log(IAP) values obtained for the 46 samples is 3.07 ± 0.34 (1σ), which is consistent with the solubility constant 3.00 ± 0.12 ( Davison et al., 1999) of amorphous FeS, implying that the anaerobic aquifers in the Choshui fan-delta are still undergoing active sulfate-reduction processes and keeping the groundwater saturated with amorphous FeS. We suggest that the -logKsp value 3.91 of amorphous FeS adopted in the databases for WATEQF and PHREEQC computer programs ought to be revised to 3.00. Otherwise, the saturation indices (SI) calculated by the two computer programs will be an order of magnitude too high.

  8. Chemical Composition and In Vitro Cytotoxic Activity of Essential Oil of Leaves of Malus domestica Growing in Western Himalaya (India)

    PubMed Central

    Walia, Mayanka; Mann, Tavleen S.; Kumar, Dharmesh; Agnihotri, Vijai K.; Singh, Bikram

    2012-01-01

    Light pale-colored volatile oil was obtained from fresh leaves of Malus domestica tree, growing in Dhauladhar range of Himalaya (Himachal Pradesh, India), with characteristic eucalyptol dominant fragrance. The oil was found to be a complex mixture of mono-, sesqui-, di-terpenes, phenolics, and aliphatic hydrocarbons. Seventeen compounds accounting for nearly 95.3% of the oil were characterized with the help of capillary GC, GC-MS, and NMR. Major compounds of the oil were characterized as eucalyptol (43.7%), phytol (11.5%), α-farnesene (9.6%), and pentacosane (7.6%). Cytotoxicity of essential oil of leaves of M. domestica was evaluated by sulforhodamine B (SRB) assays. The essential oil of leaves of M. domestica, tested against three cancer cell lines, namely, C-6 (glioma cells), A549 (human lung carcinoma), CHOK1 (Chinese hamster ovary cells), and THP-1 (human acute monocytic leukemia cell). The highest activity showed by essential oil on C-6 cell lines (98.2%) at concentration of 2000 μg/ml compared to control. It is the first paper in literature to exploit the chemical composition and cytotoxic activity of leaves essential oil of M. domestica. PMID:22619691

  9. 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.

  10. Anatomy of an earthquake multiplet active over several years in the western part of the Corinth rift

    NASA Astrophysics Data System (ADS)

    Godano, Maxime; Bernard, Pascal; Marsan, David; Dublanchet, Pierre

    2014-05-01

    The Corinth rift is one of the most seismically zones in Europe. The seismic activity is characterized by numerous multiplets. A multiplet is a group of earthquakes with similar waveforms resulting from close location and focal mechanism. Multiplets are often associated with small asperities and can be seen as repeated ruptures due to transient forcing as silent creep or pore pressure front diffusion. Detailed analysis of the multiplets in the Corinth rift is an opportunity to better understand fault dynamics, small earthquake rupture mechanics and coupling with aseismic processes. We focus on a large multiplet (500 x 500 m) located under the northern coast of the Corinth gulf at 8 km depth. This multiplet was more or less regularly active between 2000 and 2007. During this period, 56 events were recorded. The most observed recurrence time is of 23 days but can vary between 1 and 115 days. We estimate the source parameters of the 56 earthquakes by following a two-step approach based on the analysis of the displacement seismic spectrum. First, the scalar seismic moment and the magnitude are computed from the amplitude of the low frequency part (plateau) of the P and S spectrum. Second the source size is calculated from the P and S corner frequencies. Corner frequencies are determined by inverting spectral ratio (i.e. the ratio between the spectra of two collocated earthquakes). The advantage of working with spectral ratio is to eliminate the trade-off between corner frequency and anelastic attenuation if Q factor is poorly known. Spectral ratio inversion is performed following a Bayesian formalism. The magnitudes scale between 1.20 and 2.76. The seismic activity is characterized by relatively high magnitude events (b-value = 0.82) until the mainshock (mid-2003) and low magnitude events after (b-value = 1.21). The source radii globally range between 50 and 200 m. The source overlapping is strong; some fault patches have ruptured up to19 times which has produced a

  11. 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

  12. 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

  13. Composition and in vitro cytotoxic activities of essential oil of Hedychium spicatum from different geographical regions of western Himalaya by principal components analysis.

    PubMed

    Mishra, Tripti; Pal, Mahesh; Meena, Sanjeev; Datta, Dipak; Dixit, Prateek; Kumar, Anil; Meena, Baleshwar; Rana, T S; Upreti, D K

    2016-01-01

    The rhizome of Hedychium spicatum has been widely used in traditional medicines. The present study deals with the evaluation of the cytotoxic potential of rhizome essential oils from four different regions of the Western Himalaya (India) along with comparative correlation analysis to characterise the bioactive cytotoxic component. The essential oils were coded as MHS-1, MHS-2, MHS-3 and MHS-4, and characterised using GC-FID and GC-MS. The main volatile compounds identified were 1,8-cineol, eudesmol, cubenol, spathulenol and α-cadinol. In vitro cytotoxic activities were assessed against human cancer cell lines such as, the lung (A549), colon (DLD-1, SW 620), breast (MCF-7, MDA-MB-231), head and neck (FaDu), and cervix (HeLa). MHS-4 is significantly active in comparison to other samples against all cancer cell lines. Sample MHS-4 has major proportion of monoterpene alcohol mainly 1,8-cineol. Principal components analysis was performed for the experimental results and all four samples were clustered according to their percentage inhibition at different doses. PMID:26196318

  14. 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.

  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. PMID:25731086

  16. 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

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

    PubMed Central

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

    2011-01-01

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

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

    PubMed Central

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

    1992-01-01

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

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

    PubMed

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

    2014-01-17

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

  20. 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

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

    PubMed

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

    2012-12-01

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

  2. 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

  3. 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

  4. The sequence of moderate-size earthquakes at the junction of the Ligurian basin and the Corsica margin (western Mediterranean): The initiation of an active deformation zone revealed?

    NASA Astrophysics Data System (ADS)

    Larroque, Christophe; Delouis, Bertrand; Sage, Françoise; Régnier, Marc; Béthoux, Nicole; Courboulex, Françoise; Deschamps, Anne

    2016-04-01

    A new seismically active zone is found in the southern part of the Ligurian basin, 80-km west of Corsica (western Mediterranean). The activity began in February 2011 with a foreshock (ML 4) and a mainshock (ML 5.3) 5 days later, followed by numerous aftershocks. We first analyze the fore- and mainshock in detail. We compare the results obtained using classical methods (linear location in a 1D medium and focal mechanisms from P and S polarities) and new approaches (non-linear location in a 3D medium and waveform modeling for determining the seismic moment and the focal mechanism). Both methods provided similar results for location, depth (in the range of 6-13 km) and focal mechanisms, which reveal reverse faulting with nodal planes oriented N-S and NE-SW. We then locate 27 of the aftershocks in the 3D model and find a 10-km-long NE-SW alignment with a depth between 7 and 16 km. In 2012 and 2013, three other moderate-size events (ML 3.8, 4 and 4.5) occurred and confirm that this zone is still active. The epicentral area is located in the oceanic domain of the Ligurian Basin. From analysis of the bathymetry and high-resolution multi-channel seismic profiles, no morphologic anomaly at surface and no inherited fault in the shallow ~ 4 km depth were imaged, which suggest that no significant deformation occurred in the area since 5 Ma. Thus, the structure(s) activated during the 2011-2013 sequence remain unknown. In light of these results, we point out a notable difference on both sides of the Ligurian Basin: the northern margin, close to the alpine chain, suffered strong earthquakes and large cumulated deformation since 5 Ma, while the southern margin, close to the Corsica-Sardinia continental block, is poorly deformed since 5 Ma.

  5. 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

  6. Kinematics and active faults in Western Greece in the framework of Central and Eastern Mediterranean geodynamics: new insights from GPS and field work

    NASA Astrophysics Data System (ADS)

    Perouse, E.; Chamot-Rooke, N. R.; Sébrier, M.; Rabaute, A.; Sorel, D.; Arsenikos, S.; Briole, P.

    2011-12-01

    Western Greece is the transition area between fast extension in the Gulf of Corinth, dextral transpression in the Ionian islands associated to the NW termination of the Hellenic subduction zone and collision of the Apulian platform and Eurasia plate in Epirus. We use a multi-scale approach to investigate the kinematics of this challenging region. As the kinematic boundary conditions of this region are complex, we propose a large scale kinematic model that encompasses both Central and Eastern Mediterranean. We use Haines and Holt (1993) method to derive a continuous velocity and strain rate field by interpolating model velocities that are fitted in a least square sense to published GPS velocities. The main results of our model are: (1) We minimize the motion of the Hyblean plateau in Sicily and Apulia to derive a pole of rotation for the Ionian block following D'Agostino et al. (2008). This rotation pole also minimizes the Ionian basin motion; (2) We propose a "southward drag" of SW Balkans associated to the retreat of the Hellenic slab. The southward motion of the Aegean block with respect to Eurasia is gradually accommodated in the Balkans, so that the propagation of the North Anatolian Fault throughout is not required; (3) On absolute plates motion reference frame (Kreemer 2009), our results suggest a surface toroidal motion located above the NW edge of the Hellenic slab. We discuss whether this clockwise surface toroidal motion mimics toroidal astenospheric flow or not. At a smaller scale, we perform kinematic calculations to distinguish whether surface movements measured by GPS in Western Greece are due to block motion or diffuse deformation. Interseismic strain influence of the shallow subduction interface on surface velocity vectors is tested. To constrain the active structures that accommodate the present strain, we carried out a Quaternary geology and geomorphology field work in Etolo-Akarnania region (Western Greece). We establish a new map of active

  7. 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.

  8. 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

  9. 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.

  10. Species Composition and Seasonal Activities of Malaria Vectors in an Area at Reintroduction Prevention Stage, Khuzestan, South-Western Iran

    PubMed Central

    Maghsoodi, Naimatallah; Ladonni, Hossin; Basseri, Hamid Reza

    2015-01-01

    Background: The most part of Iran become malaria-free region and fall in prevention of re-introduction stage. These regions however are struggling with imported of malaria cases where malaria vectors exist. Therefore, understanding the situation of mosquito vectors is crucial. This study was carried out to find out the present situation of malaria vectors and malaria transmission potential in a malaria-free area. Methods: The study was conducted in a malaria free area, Izeh County, Khuzestan Province during 12 months in 2011–2012. Five villages, including 2 in highlands and 3 in plain area, were selected randomly. The mosquito sampling methods were conducted using spray sheet and hand catch collection methods from indoor/outdoors, window trap and larvae collections. Results: In total, 3352 female Anopheles were captured, 1826 mosquito from highland and 1526 from plain areas. Five species, An. stephensi, An. fluviatilis s.l., An. dthali, An. superpictus and An. pulcherrimus were identified. The seasonal activities were started from April to March. The abdominal conditions of collected mosquitoes from indoor/outdoor places pointed to exophilic propensity of An. fluviatilis.l. s.l. and endophilic behaviour for rest of the vectors. The results of window trap also confirmed these behaviors. The larval habitats of four species were widely dispersed and included spring, margin of rivers, irrigation channels, stagnant water and rice filed. Conclusion: Understanding the present situation of malaria vectors in free-malaria area is crucial particularly where is struggling with imported cases. The results of present study can be expanded to other area of northern Khuzestan for malaria vector control planning in reintroduction prevention stage. PMID:26114144

  11. 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...

  12. 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

  13. 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

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

    PubMed Central

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

    2014-01-01

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

  15. UAV-based photogrammetry combination of the elevational outcrop and digital surface models: an example of Sanyi active fault in western Taiwan

    NASA Astrophysics Data System (ADS)

    Hsieh, Cheng-En; Huang, Wen-Jeng; Chang, Ping-Yu; Lo, Wei

    2016-04-01

    An unmanned aerial vehicle (UAV) with a digital camera is an efficient tool for geologists to investigate structure patterns in the field. By setting ground control points (GCPs), UAV-based photogrammetry provides high-quality and quantitative results such as a digital surface model (DSM) and orthomosaic and elevational images. We combine the elevational outcrop 3D model and a digital surface model together to analyze the structural characteristics of Sanyi active fault in Houli-Fengyuan area, western Taiwan. Furthermore, we collect resistivity survey profiles and drilling core data in the Fengyuan District in order to build the subsurface fault geometry. The ground sample distance (GSD) of an elevational outcrop 3D model is 3.64 cm/pixel in this study. Our preliminary result shows that 5 fault branches are distributed 500 meters wide on the elevational outcrop and the width of Sanyi fault zone is likely much great than this value. Together with our field observations, we propose a structural evolution model to demonstrate how the 5 fault branches developed. The resistivity survey profiles show that Holocene gravel was disturbed by the Sanyi fault in Fengyuan area.

  16. [Quantitative Analysis of Immuno-fluorescence of Nuclear Factor-κB Activation].

    PubMed

    Xiu, Min; He, Feng; Lou, Yuanlei; Xu, Lu; Xiong Jieqi; Wang, Ping; Liu, Sisun; Guo, Fei

    2015-06-01

    Immuno-fluorescence technique can qualitatively determine certain nuclear translocation, of which NF-κB/ p65 implicates the activation of NF-κB signal pathways. Immuno-fluorescence analysis software with independent property rights is able to quantitatively analyze dynamic location of NF-κB/p65 by computing relative fluorescence units in nuclei and cytoplasm. We verified the quantitative analysis by Western Blot. When we applied the software to analysis of nuclear translocation in lipopolysaccharide (LPS) induced (0. 5 h, 1 h, 2 h, 4 h) primary human umbilical vein endothelial cells (HUVECs) , we found that nuclear translocation peak showed up at 2h as with calculated Western blot verification results, indicating that the inventive immuno-fluorescence analysis software can be applied to the quantitative analysis of immuno-fluorescence. PMID:26485997

  17. Analysis of glabrous canary seeds by ELISA, mass spectrometry, and Western blotting for the absence of cross-reactivity with major plant food allergens.

    PubMed

    Boye, Joyce Irene; Achouri, Allaoua; Raymond, Nancy; Cleroux, Chantal; Weber, Dorcas; Koerner, Terence B; Hucl, Pierre; Patterson, Carol Ann

    2013-06-26

    Glabrous (hairless) canary seed belongs to the Poaceae (Gramineae) family and could serve as an alternative source of gluten-free cereal grain. In this study, allergenic cross-reactivities between hairless, dehulled canary seeds (Phalaris canariensis) and major allergenic proteins from gluten, soy, peanuts, tree nuts, sesame, and mustard were studied using commercial enzyme-linked immune sorbent assay (ELISA) kits specific for these target allergens. Mass spectrometry (MS) and immunoblotting were further used to assess for the presence of gluten-specific protein fragments. MS results revealed the likely presence of proteins homologous with rice, oat, corn, carrot, tomato, radish, beet, and chickpea. However, no presence of celiac-related gluten fragments from wheat, rye, barley, or their derivatives was found. Immunoblotting studies yielded negative results, further confirming the absence of gluten in the canary seed samples tested. No cross-reactivities were detected between canary seeds and almond, hazelnut, mustard, peanut, sesame, soy, walnut, and gluten using ELISA. PMID:23706175

  18. 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...

  19. Cross-Reactivity Pattern of Asian and American Human Gnathostomiasis in Western Blot Assays Using Crude Antigens Prepared from Gnathostoma spinigerum and Gnathostoma binucleatum Third-Stage Larvae.

    PubMed

    Neumayr, Andreas; Ollague, Jose; Bravo, Francisco; Gotuzzo, Eduardo; Jimenez, Pedro; Norton, Scott A; Doanh, Pham Ngoc; Nawa, Yukifumi; Horii, Yoichiro; Nickel, Beatrice; Marti, Hanspeter

    2016-08-01

    Gnathostomiasis is a zoonotic parasitosis endemic in many Asian and some Latin American countries. Most human infections are caused by Gnathostoma spinigerum in Asia and Gnathostoma binucleatum in the Americas, and recently, imported cases have been increasing among travelers returning from endemic regions. Confirmation of the clinical diagnosis relies largely on serologic tests, with a G. spinigerum-antigen-based immunoblot currently being the diagnostic method of choice. However, we repeatedly experienced that sera from patients with clinically suspected American gnathostomiasis gave negative results in this assay. Therefore, we used homologous methods to prepare G. spinigerum- and G. binucleatum-antigen-based immunoblot assays, and evaluated the cross-reactivity of the two assays. The results show incomplete cross-reactivity between the two assays: the G. spinigerum-antigen-based immunoblot apparently only detects Asian gnathostomiasis caused by G. spinigerum, whereas the G. binucleatum-antigen-based immunoblot is apparently capable of detecting American as well as Asian gnathostomiasis. PMID:27325806

  20. 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…

  1. 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

  2. Astragalin inhibits IL-1β-induced inflammatory mediators production in human osteoarthritis chondrocyte by inhibiting NF-κB and MAPK activation.

    PubMed

    Ma, Zhiqiang; Piao, Taikui; Wang, Yanlong; Liu, Jianyu

    2015-03-01

    Astragalin, a bioactive component isolated from Rosa agrestis, has been described to exhibit anti-inflammatory activity. The aim of this study was to investigate the anti-inflammatory effects and the underlying mechanisms of astragalin on IL-1β-stimulated human osteoarthritis chondrocyte. The production of NO and PGE2 was detected by Griess reaction and ELISA. The expression of iNOS and COX-2 was detected by western blotting. The expression of NF-κB and MAPKs was detected by western blot analysis. We found that astragalin dose-dependently inhibited IL-1β-induced NO and PGE2 production, as well as iNOS and COX-2 expression. Meanwhile, western blot analysis showed that astragalin inhibited IL-1β-induced NF-κB and MAPK activation in human osteoarthritis chondrocyte. In addition, astragalin was found to activate PPAR-γ. The inhibition of astragalin on IL-1β-induced NO and PGE2 production can be reversed by PPAR-γ antagonist GW9662. Astragalin suppressed IL-1β-induced inflammatory mediators via activating PPAR-γ, which subsequently inhibited IL-1β-induced NF-κB and MAPK activation. Astragalin may be a potential agent in the treatment of osteoarthritis. PMID:25637445

  3. Mitochondrial quality control, promoted by PGC-1α, is dysregulated by Western diet-induced obesity and partially restored by moderate physical activity in mice

    PubMed Central

    Greene, Nicholas P; Lee, David E; Brown, Jacob L; Rosa, Megan E; Brown, Lemuel A; Perry, Richard A; Henry, Jordyn N; Washington, Tyrone A

    2015-01-01

    Skeletal muscle mitochondrial degeneration is a hallmark of insulin resistance/obesity marked by lost function, enhanced ROS emission, and altered morphology which may be ameliorated by physical activity (PA). However, no prior report has examined mitochondrial quality control regulation throughout biogenesis, fusion/fission dynamics, autophagy, and mitochondrial permeability transition pore (MPTP) in obesity. Therefore, we determined how each process is impacted by Western diet (WD)-induced obesity and whether voluntary PA may alleviate derangements in mitochondrial quality control mechanisms. Despite greater mitochondrial content following WD (COX-IV and Cytochrome C), induction of biogenesis controllers appears impaired (failed induction of PGC-1α). Mitochondrial fusion seems diminished (reduced MFN2, Opa1 proteins), with no significant changes in fission, suggesting a shift in balance of dynamics regulation favoring fission. Autophagy flux was promoted in WD (reduced p62, increased LC3II:I ratio); however, mitophagy marker BNIP3 is reduced in WD which may indicate reduced mitophagy despite enhanced total autophagy flux. MPTP regulator Ant mRNA is reduced by WD. Few processes were impacted by physical activity. Finally, mitochondrial quality control processes are partially promoted by PGC-1α, as PGC-1α transgenic mice display elevated mitochondrial biogenesis and autophagy flux. Additionally, these mice exhibit elevated Mfn1 and Opa1 mRNA, with no change in protein content suggesting these factors are transcriptionally promoted by PGC-1α overexpression. These data demonstrate dysfunctions across mitochondrial quality control in obesity and that PGC-1α is sufficient to promote multiple, but not necessarily all, aspects of mitochondrial quality control. Mitochondrial quality control may therefore be an opportune target to therapeutically treat metabolic disease. PMID:26177961

  4. 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

  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

    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

  6. Mitochondrial quality control, promoted by PGC-1α, is dysregulated by Western diet-induced obesity and partially restored by moderate physical activity in mice.

    PubMed

    Greene, Nicholas P; Lee, David E; Brown, Jacob L; Rosa, Megan E; Brown, Lemuel A; Perry, Richard A; Henry, Jordyn N; Washington, Tyrone A

    2015-07-01

    Skeletal muscle mitochondrial degeneration is a hallmark of insulin resistance/obesity marked by lost function, enhanced ROS emission, and altered morphology which may be ameliorated by physical activity (PA). However, no prior report has examined mitochondrial quality control regulation throughout biogenesis, fusion/fission dynamics, autophagy, and mitochondrial permeability transition pore (MPTP) in obesity. Therefore, we determined how each process is impacted by Western diet (WD)-induced obesity and whether voluntary PA may alleviate derangements in mitochondrial quality control mechanisms. Despite greater mitochondrial content following WD (COX-IV and Cytochrome C), induction of biogenesis controllers appears impaired (failed induction of PGC-1α). Mitochondrial fusion seems diminished (reduced MFN2, Opa1 proteins), with no significant changes in fission, suggesting a shift in balance of dynamics regulation favoring fission. Autophagy flux was promoted in WD (reduced p62, increased LC3II:I ratio); however, mitophagy marker BNIP3 is reduced in WD which may indicate reduced mitophagy despite enhanced total autophagy flux. MPTP regulator Ant mRNA is reduced by WD. Few processes were impacted by physical activity. Finally, mitochondrial quality control processes are partially promoted by PGC-1α, as PGC-1α transgenic mice display elevated mitochondrial biogenesis and autophagy flux. Additionally, these mice exhibit elevated Mfn1 and Opa1 mRNA, with no change in protein content suggesting these factors are transcriptionally promoted by PGC-1α overexpression. These data demonstrate dysfunctions across mitochondrial quality control in obesity and that PGC-1α is sufficient to promote multiple, but not necessarily all, aspects of mitochondrial quality control. Mitochondrial quality control may therefore be an opportune target to therapeutically treat metabolic disease. PMID:26177961

  7. 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.

  8. 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.

  9. 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

  10. 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

  11. 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

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

    PubMed

    Tonosaki, K; Nishio, Takeshi

    2010-10-01

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

  13. Differential river incision across active normal faults in Grand Canyon: a response to mantle-driven uplift of the western Colorado Plateau

    NASA Astrophysics Data System (ADS)

    Karlstrom, K. E.; Crow, R.

    2010-12-01

    High-quality gravel-constrained incision rates have been calculated in western Grand Canyon in a reach between river mile (RM) 177 and 246 where 100-840 ka basalt flows erupted and cascaded into Grand Canyon and flowed downriver over 120 km. Bedrock incision rates are calculated at numerous locations where dated basalts overly gravel-capped bedrock straths; fault slip is measured via Ar-Ar dating of offset basalts. Marked variations in incision rate take place across the active Hurricane and Toroweap normal faults, with incision rates in the footwall of the faults being subequal to the incision rate in the immediate hanging-wall plus the fault slip rate. However incision rates increase progressively away from the fault due to formation of a hanging-wall anticline. Nine new and updated incision rates based on new 40Ar/39Ar analyses from Grand Canyon’s intra-canyon basalts support and extend earlier conclusions that: 1) incision rate discrepancies across active faults are explained by fault dampening of incision rates, 2) incision rate variations in the fault block between the Toroweap and the Hurricane faults shows a hanging-wall anticline with wavelength < 5 km. 3) large-scale ~100 m/Ma discrepancy in incision between eastern (~175 m/Ma) and western Grand Canyon (~65 m/Ma) across the Hurricane fault may reflect broad mantle-driven epeirogenic surface uplift of the Colorado Plateau. This last hypothesis is supported by profile reconstructions and geodynamic models. Postulated sub-equal incision across the eastern Grand Canyon block, which would support epeirogenic uplift, are being further tested using U-Series dating of high terrace flights at RM 65, 69, 95, and 116. Neogene surface uplift of eastern Grand Canyon and the margins of the Colorado Plateau is consistent with: 1) high elevations along the plateau margins, 2) geoid anomalies, 3) tomographic and magnetotelluric data showing low-velocity conductive mantle under plateau margins, 4) temporal and spatial

  14. 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.

  15. 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

  16. Characterizing the mechanism of action of double-stranded RNA activity against western corn rootworm (Diabrotica virgifera virgifera LeConte).

    PubMed

    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

  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. 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

  19. 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

  20. 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

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

    PubMed

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

    1993-11-01

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

  2. 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

  3. 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

  4. Resistance evolution to the first generation of genetically modified Diabrotica-active Bt-maize events by western corn rootworm: management and monitoring considerations

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 (GM) Bt-maize offers an additional means of control against W...

  5. The Influence of Feral Horse Activity on Water and Shellfish (Gukensia demissa) Quality Along the Western Coast of Assateague Island National Seashore, Maryland

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Feral horses (Equus caballus) inhabit portions of the Western United States and some barrier islands along the East Coast. Approximately 150 feral horses are located on Assateague Island National Seashore (ASIS), Maryland, a barrier island popular with tourists and recreational fishermen. This stu...

  6. 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…

  7. Measurement of constitutive MAPK and PI3K/AKT signaling activity in human cancer cell lines

    PubMed Central

    Paraiso, Kim H.T.; Van Der Kooi, Kaisa; Messina, Jane L.; Smalley, Keiran S. M.

    2014-01-01

    The growth and survival of cancer cells is often driven by constitutive activity in the mitogen activated protein kinase (MAPK) and phospho-inositide 3-kinase (PI3K)/AKT signaling pathways. Activity in these signal transduction cascades is known to contribute to the uncontrolled growth and resistance to apoptosis that characterizes tumor progression. There is now a great deal of interest in therapeutically targeting these pathways in cancer using small molecule inhibitors. In this chapter we describe methods to measure constitutive MAPK and AKT activity in melanoma cell lines, with a focus upon Western blotting, phospho-flow cytometry and immunofluorescence staining techniques. PMID:21036250

  8. Robot Printing of Reverse Dot Blot Arrays for Human Mutation Detection

    PubMed Central

    Lappin, Stephen; Cahlik, Jeff; Gold, Bert

    2001-01-01

    We report on a generally useful, partially automated, human mutation detection method based upon printing moderate density oligonucleotide arrays using a biorobot on activated nylon membranes. The Beckman Biomek 2000 was adapted to this task through fabrication of aluminum membrane filter holders and the development of an addressable Tool Command Language (Tcl) program, which can be invoked through BioScript. During program execution, a robot arm is moved along the x, y, and z axes to expel liquid, without dripping, from disposable barrier pipette tips and then to touch the drops on preactivated membranes. Printed arrays consist of alternating rows of oligonucleotides containing normal and mutant sequences. Hybridization of biotin labeled polymerase chain reaction products derived from human patient genomic DNA samples are visualized using chemiluminescent or chromogenic indicators. This technique allows unequivocal genotyping of 32 mutations at the β-thalassemia locus (11p15.5) and of 34 mutations and one polymorphism at the cystic fibrosis transconductance membrane regulator locus (7p35). PMID:11687602

  9. 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

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

    PubMed

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

    1997-12-01

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

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

    SciTech Connect

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

    1988-05-01

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

  12. Studies on prophenoloxidase activation in the mosquito Aedes aegypti L.

    PubMed

    Ashida, M; Kinoshita, K; Brey, P T

    1990-03-30

    This study, the first of its kind in a mosquito vector species, demonstrates the feasibility of studying prophenoloxidase activation in an insect containing not more than a few microliters of hemolymph. Mosquito phenoloxidase was found to be in an inactive proenzyme form, prophenoloxidase. Mosquito prophenoloxidase required bivalent cation for its activation; Ca2+ was found to be the most efficient for activation. Concomitant amidase activity was also observed prior to phenoloxidase activity. Through Western blotting, using a cross-reactive silkworm antiprophenoloxidase antibody, our results strongly suggest that mosquito prophenoloxidase activation resulted from limited proteolysis. Protease inhibitor studies reinforced this contention showing the involvement of (a) serine protease(s) with trypsin-like activity in the activation of mosquito prophenoloxidase. PMID:2110057

  13. Detection of immunogenic parasite and host-specific proteins in the sera of active and chronic individuals infected with Toxoplasma gondii.

    PubMed

    Yeng, Chen; Osman, Emelia; Mohamed, Zeehaida; Noordin, Rahmah

    2010-12-01

    Toxoplasma gondii infection in pregnant women may result in abortion and foetal abnormalities, and may be life-threatening in immunocompromised hosts. To identify the potential infection markers of this disease, 2-DE and Western blot methods were employed to study the parasite circulating antigens and host-specific proteins in the sera of T. gondii-infected individuals. The comparisons were made between serum protein profiles of infected (n=31) and normal (n=10) subjects. Antigenic proteins were identified by immunoblotting using pooled sera and monoclonal anti-human IgM-HRP. Selected protein spots were characterised using mass spectrometry. Prominent differences were observed when serum samples of T. gondii-infected individuals and normal controls were compared. A significant up-regulation of host-specific proteins, α(2)-HS glycoprotein and α(1)-B glycoprotein, was also observed in the silver-stained gels of both active and chronic infections. However, only α(2)-HS glycoprotein and α(1)-B glycoprotein in the active infection showed immunoreactivity in Western blots. In addition, three spots of T. gondii proteins were detected, namely (i) hypothetical protein chrXII: 3984434-3 TGME 49, (ii) dual specificity protein phosphatase, catalytic domain TGME 49 and (iii) NADPH-cytochrome p450 reductase TGME 49. Thus, 2-DE approach followed by Western blotting has enabled the identification of five potential infection markers for the diagnosis of toxoplasmosis: three are parasite-specific proteins and two are host-specific proteins. PMID:21080484

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

    SciTech Connect

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

    1994-09-01

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

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

    PubMed Central

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

    2014-01-01

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

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

    PubMed

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

    2014-01-01

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

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

    PubMed

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

    2011-02-01

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

  18. 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.

  19. 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…

  20. 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.

  1. 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

  2. Activating Peroxisome Proliferator-Activated Receptors (PPARs): a New Sight for Chrysophanol to Treat Paraquat-Induced Lung Injury.

    PubMed

    Li, Ang; Liu, Yuguang; Zhai, Lu; Wang, Liying; Lin, Zhe; Wang, Shumin

    2016-04-01

    The aim of this study is to evaluate the protective effects of chrysophanol (CH) against paraquat (PQ)-induced pulmonary injury. Fifty BALB/C mice were randomized into five groups: (1) control, (2) PQ, (3) PQ + dexamethasone (Dex, 2 mg/kg), (4) PQ + CH (10 mg/kg), and (5) PQ + CH (20 mg/kg). A single dose of PQ (50 mg/kg, i.p.) was intraperitoneally given to induce acute lung injury. Then mice were treated with CH (10 and 20 mg/kg/day, orally) for 7 days. At the end of the experiment, animals were euthanized and then bronchoalveolar lavage fluid (BALF) and lung tissues were collected for histological observation, biochemical analysis, and Western blot analysis. Malondialdehyde (MDA), myeloperoxidase (MPO), superoxide dismutase (SOD), interleukin-6 (IL-6), IL-1β, and tumor necrosis factor-α (TNF-α) levels in BALF were determined. The levels of SOD and MDA in the lung were also detected. The peroxisome proliferator-activated receptor (PPAR)-γ and nuclear factor-kappaB (NF-κB) pathway proteins in the lung were determined by Western blot. Histological examination indicated that CH attenuated lung inflammation caused by PQ. Biochemical results showed that CH treatment significantly reduced the levels of MDA, MPO, and inflammatory cytokines and increased the level of SOD, compared to those in the PQ group. Meanwhile, Western Blot results revealed that CH increased PPAR-γ expression and inhibited NF-κB pathway activation after PQ challenge. These findings suggested the potential therapeutic effects of CH which is derived from a natural product on PQ-induced pulmonary injury. PMID:26920845

  3. 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

  4. [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

  5. 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.

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

    SciTech Connect

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

    1988-08-25

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

  7. Melatonin alleviates myosin light chain kinase expression and activity via the mitogen-activated protein kinase pathway during atherosclerosis in rabbits.

    PubMed

    Cheng, Xiaowen; Wan, Yufeng; Xu, Yuanhong; Zhou, Qing; Wang, Yuan; Zhu, Huaqing

    2015-01-01

    Melatonin (MLT) is an endogenous indole compound with numerous biological activities that has been associated with atherosclerosis (AS). In the present study, rabbits were used as an AS model in order to investigate whether MLT affects endothelial cell permeability, myosin light chain kinase (MLCK) activity and MLCK expression via the mitogen-activated protein kinase (MAPK) pathway. Expression and activity of MLCK were measured using western blot analysis, quantitative polymerase chain reaction, immunohistochemistry and γ-32P-adenosine triphosphate incorporation. Endothelial permeability was detected using rhodamine phalloidin fluorescence staining. The phosphorylation of extracellular regulated protein kinase (ERK), c-Jun N-terminal kinase (JNK) and p38 in endothelial cells were also analyzed using western blot analysis. Atheromatous plaques were formed in rabbits with a high cholesterol diet; however, following treatment with MLT, the number and areas of atheromatous plaques were significantly reduced. In addition, MLT treatment reversed the increase of MLCK activity and expression that occurred in rabbits with high cholesterol intake. Furthermore, levels of phosphorylated ERK, JNK and p38 decreased following MLT treatment. In conclusion, the results of the present study indicated that AS may be associated with increased MLCK expression and activity, which was reduced following treatment with MLT. The mechanism of action of MLT was thought to proceed via modulating MAPK pathway signal transduction; however, further studies are required in order to fully elucidate the exact regulatory mechanisms involved. PMID:25339116

  8. A peptide derived from phage display library exhibits anti-tumor activity by targeting GRP78 in gastric cancer multidrug resistance cells.

    PubMed

    Kang, Jianqin; Zhao, Guohong; Lin, Tao; Tang, Shanhong; Xu, Guanghui; Hu, Sijun; Bi, Qian; Guo, Changcun; Sun, Li; Han, Shuang; Xu, Qian; Nie, Yongzhan; Wang, Biaoluo; Liang, Shuhui; Ding, Jie; Wu, Kaichun

    2013-10-10

    Multidrug resistance (MDR) remains a significant challenge to the clinical treatment of gastric cancer (GC). In the present study, using a phage display approach combined with MTT assays, we screened a specific peptide GMBP1 (Gastric cancer MDR cell-specific binding peptide), ETAPLSTMLSPY, which could bind to the surface of GC MDR cells specifically and reverse their MDR phenotypes. Immunocytochemical staining showed that the potential receptor of GMBP1 was located at the membrane and cytoplasm of MDR cells. In vitro and in vivo drug sensitivity assays, FACS analysis and Western blotting confirmed that GMBP1 was able to re-sensitize MDR cells to chemical drugs. Western blotting and proteomic approaches were used to screen the receptor of GMBP1, and GRP78, a MDR-related protein, was identified as a receptor of GMBP1. This result was further supported by immunofluoresence microscopy and Western blot. Additionally, Western blotting demonstrated that pre-incubation of GMBP1 in MDR cells greatly diminished MDR1, Bcl-2 and GRP78 expression but increased the expression of Bax, whereas downregulation of GRP78, function as a receptor and directly target for GMBP1, only inhibited MDR1 expression. Our findings suggest that GMBP1 could re-sensitize GC MDR cells to a variety of chemotherapeutic agents and this role might be mediated partly through down-regulating GRP78 expression and then inhibiting MDR1 expression. These findings indicate that peptide GMBP1 likely recognizes a novel GRP78 receptor and mediates cellular activities associated with the MDR phenotype, which provides new insight into research on the management of MDR in gastric cancer cells. PMID:23792224

  9. 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

  10. 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

  11. 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.

  12. Apoxin I, a novel apoptosis-inducing factor with L-amino acid oxidase activity purified from Western diamondback rattlesnake venom.

    PubMed

    Torii, S; Naito, M; Tsuruo, T

    1997-04-01

    Venom of the western diamondback rattlesnake (Crotalus atrox) induces apoptosis in human umbilical vein endothelial cells, which could result in hemorrhage in tissues bitten by the snake. To identify the hemorrhagic factor, we purified a novel protein, apoxin I, from rattlesnake venom. Apoxin I induced apoptosis in human umbilical vein endothelial, human promyelocytic leukemia HL-60, human ovarian carcinoma A2780, and mouse endothelial KN-3 cells. Amino acid sequence analysis of the apoxin I showed close similarity to L-amino acid oxidase from the Malayan pit viper (Calloselasma rhodostoma). The purified apoxin I oxidized L-leucine but not D-leucine to produce H2O2. The apoxin I-induced apoptosis was inhibited by catalase, a H2O2 scavenger. These results indicate that the H2O2 produced by L-amino acid oxidation by apoxin I is involved in the apoxin I-induced apoptosis and in hemorrhage caused by rattlesnake venom. PMID:9083096

  13. Down-Regulation of Telomerase Activity and Activation of Caspase-3 Are Responsible for Tanshinone I-Induced Apoptosis in Monocyte Leukemia Cells in Vitro

    PubMed Central

    Liu, Xiao-Dan; Fan, Rui-Fang; Zhang, Yong; Yang, Hong-Zhi; Fang, Zhi-Gang; Guan, Wei-Bing; Lin, Dong-Jun; Xiao, Ruo-Zhi; Huang, Ren-Wei; Huang, He-Qing; Liu, Pei-Qing; Liu, Jia-Jun

    2010-01-01

    Tanshinone I (Tan-I) is a diterpene quinone extracted from the traditional herbal medicine Salvia miltiorrhiza Bunge. Recently, Tan-I has been reported to have anti-tumor effects. In this study, we investigated the growth inhibition and apoptosis inducing effects of Tan-I on three kinds of monocytic leukemia cells (U937, THP-1 and SHI 1). Cell viability was measured by MTT assay. Cell apoptosis was assessed by flow cytometry (FCM) and AnnexinV/PI staining. Reverse transcriptase polymerase chain reaction (RT-PCR) and PCR–enzyme-linked immunosorbent assay (ELISA) were used to detect human telomerase reverse transcriptase (hTERT) expression and telomerase activity before and after apoptosis. The activity of caspase-3 was determined by Caspase colorimetric assay kit and Western blot analysis. Expression of the anti-apoptotic gene Survivin was assayed by Western blot and Real-time RT-PCR using the ABI PRISM 7500 Sequence Detection System. The results revealed that Tan-I could inhibit the growth of these three kinds of leukemia cells and cause apoptosis in a time- and dose-dependent manner. After treatment by Tan-I for 48 h, Western blotting showed cleavage of the caspase-3 zymogen protein with the appearance of its 17-kD subunit, and a 89-kD cleavage product of poly (ADP-ribose) polymerase (PARP), a known substrate of caspase-3, was also found clearly. The expression of hTERT mRNA as well as activity of telomerase were decreased concurrently in a dose-dependent manner. Moreover, Real-time RT-PCR and Western blot revealed a significant down-regulation of Survivin. We therefore conclude that the induction of apoptosis by Tan-I in monocytic leukemia U937 THP-1 and SHI 1 cells is highly correlated with activation of caspase-3 and decreasing of hTERT mRNA expression and telomerase activity as well as down-regulation of Survivin expression. To our knowledge, this is the first report about the effects of Tan-I on monocytic leukemia cells. PMID:20640151

  14. Presence and Seeding Activity of Pathological Prion Protein (PrPTSE) in Skeletal Muscles of White-Tailed Deer Infected with Chronic Wasting Disease

    PubMed Central

    Daus, Martin L.; Breyer, Johanna; Wagenfuehr, Katja; Wemheuer, Wiebke M.; Thomzig, Achim; Schulz-Schaeffer, Walter J.; Beekes, Michael

    2011-01-01

    Chronic wasting disease (CWD) is a contagious, rapidly spreading transmissible spongiform encephalopathy (TSE), or prion disease, occurring in cervids such as white tailed-deer (WTD), mule deer or elk in North America. Despite efficient horizontal transmission of CWD among cervids natural transmission of the disease to other species has not yet been observed. Here, we report for the first time a direct biochemical demonstration of pathological prion protein PrPTSE and of PrPTSE-associated seeding activity, the static and dynamic biochemical markers for biological prion infectivity, respectively, in skeletal muscles of CWD-infected cervids, i. e. WTD for which no clinical signs of CWD had been recognized. The presence of PrPTSE was detected by Western- and postfixed frozen tissue blotting, while the seeding activity of PrPTSE was revealed by protein misfolding cyclic amplification (PMCA). Semi-quantitative Western blotting indicated that the concentration of PrPTSE in skeletal muscles of CWD-infected WTD was approximately 2000-10000 -fold lower than in brain tissue. Tissue-blot-analyses revealed that PrPTSE was located in muscle-associated nerve fascicles but not, in detectable amounts, in myocytes. The presence and seeding activity of PrPTSE in skeletal muscle from CWD-infected cervids suggests prevention of such tissue in the human diet as a precautionary measure for food safety, pending on further clarification of whether CWD may be transmissible to humans. PMID:21483771

  15. Western Disturbances: A review

    NASA Astrophysics Data System (ADS)

    Dimri, A. P.; Niyogi, D.; Barros, A. P.; Ridley, J.; Mohanty, U. C.; Yasunari, T.; Sikka, D. R.

    2015-06-01

    Cyclonic storms associated with the midlatitude Subtropical Westerly Jet (SWJ), referred to as Western Disturbances (WDs), play a critical role in the meteorology of the Indian subcontinent. WDs embedded in the southward propagating SWJ produce extreme precipitation over northern India and are further enhanced over the Himalayas due to orographic land-atmosphere interactions. During December, January, and February, WD snowfall is the dominant precipitation input to establish and sustain regional snowpack, replenishing regional water resources. Spring melt is the major source of runoff to northern Indian rivers and can be linked to important hydrologic processes from aquifer recharge to flashfloods. Understanding the dynamical structure, evolution-decay, and interaction of WDs with the Himalayas is therefore necessary to improve knowledge which has wide ranging socioeconomic implications beyond short-term disaster response including cold season agricultural activities, management of water resources, and development of vulnerability-adaptive measures. In addition, WD wintertime precipitation provides critical mass input to existing glaciers and modulates the albedo characteristics of the Himalayas and Tibetan Plateau, affecting large-scale circulation and the onset of the succeeding Indian Summer Monsoon. Assessing the impacts of climate variability and change on the Indian subcontinent requires fundamental understanding of the dynamics of WDs. In particular, projected changes in the structure of the SWJ will influence evolution-decay processes of the WDs and impact Himalayan regional water availability. This review synthesizes past research on WDs with a perspective to provide a comprehensive assessment of the state of knowledge to assist both researchers and policymakers, and context for future research.

  16. Competitive interstate taxation of western coal

    SciTech Connect

    Kolstad, C.D.; Wolak, F.A. Jr.

    1983-01-01

    This paper analyzes the potential market power of western states in setting coal severance taxes. An attempt to determine the emphasis placed by the western states on the development of their coal resources is also made. Three market structures are analyzed. One involves a western regional cartel, setting taxes collectively. The other cases are noncooperative tax equilibria with Montana and Wyoming competing against each other. We study the effects on these equilibria of changes in each region's relative emphasis on development of coal resources vs tax revenue. The welfare impacts of these tax setting policies are also addressed. The analysis is based on an activity analysis of US coal markets. The results show that the taxes associated with the noncooperative competitive tax equilibria are close to present tax levels. Additionally, we conclude that western states currently are quite efficient extractors of economic rent from coal produced within their boundaries, in terms of welfare loss per dollar of tax revenue collected. 2 figures.

  17. 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, Emmanuelle; Arnaud, Nicolas; Guivel, Christèle; Lagabrielle, Yves; Scalabrino, Bruno; Espinoza, Felipe

    2010-05-01

    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.

  18. Seasonal variation in antioxidative responses and acetylcholinesterase activity in Perna viridis in eastern oceanic and western estuarine waters of Hong Kong

    NASA Astrophysics Data System (ADS)

    Lau, P. S.; Wong, H. L.; Garrigues, Ph.

    2004-10-01

    A year-round study was conducted to assess the seasonal variations and potential influence of the riverine discharge from the Pearl River on biomarker responses in Hong Kong waters. A suite of biomarkers including antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), the lipid peroxidation product malondialdehyde (MDA), a Phase II detoxification enzyme glutathione-S-transferase (GST) and the neural transmitter enzyme acetylcholinesterase (AChE) in the green mussel, Perna viridis, were monitored from three coastal sites, Port Shelter, Tung Chung and Tai O, stretching from the east to the west of Hong Kong. Despite of the seasonal variations, the total protein profiles suggested that mussels from the three sites had a growth cycle that was in phase with each other. This implied that intrinsic variation between sites due to a different phase of growth was minimal. Seasonal variations of the biomarker responses in the mussels were found to be significant (Tukey multiple comparison test, p<0.05) with a summer minimum and winter maximum. On top of seasonal variations, the western site, Tai O, was further subjected to the reduced salinity effect of the Pearl River discharge in the summer wet season. This was demonstrated by the significant July minimum in all the biomarker responses at Tai O in relation to the extreme low salinity of 8‰. Mussels from the western site also revealed a higher oxidative stress than those from the eastern side throughout the year (Tukey multiple comparison test, p<0.05), which could be caused by chemical pollutants from the Pearl River discharge. ANOVAs of the year-round dataset suggested that size was a minor factor in affecting the biomarker responses. Gill tissues of the mussels were more advantageous for biomarker studies or monitoring because their protein levels were less sensitive to seasonal variations and they yielded a higher protein normalized biomarker response than the whole body tissues. This increases their

  19. Chemical Composition and Biological Activity of Essential Oils from Wild Growing Aromatic Plant Species of Skimmia laureola and Juniperus macropoda from Western Himalaya.

    PubMed

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

    2015-06-01

    The Himalayan region is very rich in a great variety of medicinal plants. In this investigation the essential oils of two selected species are described for their antimicrobial and larvicidal as well as biting deterrent activities. Additionally, the odors are characterized. Analyzed by simultaneous GC-MS and GC-FID, the essential oils' chemical compositions are given. The main components of Skimmia laureola oil were linalool and linalyl acetate whereas sabinene was found as the main compound for Juniperus macropoda essential oil. Antibacterial testing by agar dilution assay revealed highest activity of S. laureola oil against all tested bacteria, followed by J. macropoda oil. Antifungal activity was evaluated against the strawberry anthracnose causing plant pathogens Colletotrichum acutatum, C. fragariae and C. gloeosporioides. Juniperus macropoda essential oil indicated higher antifungal activity against all three pathogens than S. laureola oil. Both essential oils showed biting deterrent activity above solvent control but low larvicidal activity. PMID:26197554

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

    PubMed

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

    2007-09-01

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

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

    PubMed Central

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

    2007-01-01

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

  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. PMID:23104293

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

    PubMed

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

    1994-08-15

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

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

    PubMed

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

    2016-08-28

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

  5. The geometry of the active strike-slip El Tigre Fault, Precordillera of San Juan, Central-Western Argentina: integrating resistivity surveys with structural and geomorphological data

    NASA Astrophysics Data System (ADS)

    Fazzito, Sabrina Y.; Cortés, José M.; Rapalini, Augusto E.; Terrizzano, Carla M.

    2013-07-01

    The geometry and related geomorphological features of the right-lateral strike-slip El Tigre Fault, one of the main morphostructural discontinuities in the Central-Western Precordillera of Argentina, were investigated. Achievements of this survey include: recognition of structural and geometrical discontinuities along the fault trace, identification and classification of landforms associated with local transpressional and transtensional sectors, observation of significant changes in the fault strike and detection of right and left bends of different wavelength. In the Central Segment of the El Tigre Fault, 2D electrical resistivity tomography surveys were carried out across the fault zone. The resistivity imaging permitted to infer the orientation of the main fault surface, the presence of blind fault branches along the fault zone, tectonic tilting of the Quaternary sedimentary cover, subsurface structure of pressure ridges and depth to the water table. Based on this information, it is possible to characterize the El Tigre Fault also as an important hydro-geological barrier. Our survey shows that the main fault surface changes along different segments from a high-angle to a subvertical setting whilst the vertical-slip component is either reverse or normal, depending on the local transpressive or transtensive regime induced by major bends along the trace. These local variations are expressed as sections of a few kilometres in length with relatively homogeneous behaviour and frequently separated by oblique or transversal structures.

  6. The effects of China's environmental protection policies on pasture quality and the activities of small-scale herders in Songpan County, Western Sichuan.

    PubMed

    Veeck, Gregory; Li, Zhou; Yu, Fawen; Emerson, Charles

    2015-06-01

    Since 2000, there has been an ambitious governmental effort to protect China's grassland resources through new, often controversial, policies and programs backed up by massive capital investments and subsidies. The policies are intended to protect pastoral ecological systems while allowing the pasture-based husbandry sector to continue to evolve in ways that assure acceptable revenues to affected families and regions. This article reports results of a case study conducted in Songpan County of western Sichuan during the fall of 2013 specifically focused on these issues and interactions. The mixed-method research combines remotely sensed MODIS vegetation i