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Sample records for activity western blot

  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. The Western Blot.

    PubMed

    Hnasko, Thomas S; Hnasko, Robert M

    2015-01-01

    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 antibody-antigen interaction and is useful for the qualitative or semiquantitative identification of specific proteins and their molecular weight from a complex mixture. This chapter will outline the requisite steps including gel electrophoresis of a protein sample, transfer of protein from a gel to a membrane support, and immunodetection of a target antigen.

  3. How to Distinguish Between the Activity of HDAC1-3 and HDAC6 with Western Blot.

    PubMed

    Beyer, Mandy; Kiweler, Nicole; Mahboobi, Siavosh; Krämer, Oliver H

    2017-01-01

    Histone deacetylases (HDACs) catalyze the deacetylation of lysine residues in their target proteins. This biochemical modification can have profound effects on the functions of these proteins and a dysregulation of HDAC activity contributes to severe diseases, including neoplastic transformation. In the following chapter, we present a strategy that allows to distinguish between the inhibition of the class I HDACs HDAC1, 2, and 3 and of the class IIb HDAC HDAC6. This method is based on Western blot and relies on the detection of hyperacetylated substrates of class I or class IIb HDACs in lysates from cells that were treated with histone deacetylase inhibitors (HDACi).

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

    PubMed

    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.

  5. Line blot and western blot immunoassays for diagnosis of Mediterranean spotted fever.

    PubMed Central

    Raoult, D; Dasch, G A

    1989-01-01

    The line blot, a new immunoassay in which antigens are placed on nitrocellulose as narrow lines, was evaluated for its sensitivity and specificity relative to the microimmunofluorescence assay for the diagnosis of Mediterranean spotted fever (MSF). The line blot assay was only slightly less sensitive and less specific than the microimmunofluorescence assay for detection of immunoglobulin M (IgM) or IgG in 100 serum specimens from 42 patients with MSF. No line blot reactions were observed among 50 control serum specimens from febrile patients with other illnesses. The line blot assay was largely group reactive for spotted fever rickettsiae, but 26% of the positive serum specimens also cross-reacted by IgM with Rickettsia typhi. Western immunoblotting was used to characterize the antigenic components recognized by 19 MSF serum specimens. For both IgM and IgG, lipopolysaccharide was the cross-reactive group antigen, whereas the high-molecular-weight species-specific protein antigens (SPAs) were the only reactive proteins. Relative to the other nine rickettsiae, Rickettsia bellii was unique both in exhibiting no SPA reactions and in having a lipopolysaccharide with a predominantly high-molecular-weight distribution. Although most of the 19 MSF serum specimens examined by Western blotting exhibited preferential reactivity to SPAs of two strains of R. conorii and weaker reactions to the other rickettsiae, 2 serum specimens exhibited SPA reactions consistent with typhus infections. In comparison with other assays, the line blot and Western blot immunoassays have advantages which may permit an improvement in the general availability and commercialization of assays for the serodiagnosis of rickettsial infections. Images PMID:2506223

  6. Western blot analysis of adhesive interactions under fluid shear conditions: the blot rolling assay.

    PubMed

    Sackstein, Robert; Fuhlbrigge, Robert

    2015-01-01

    Western blotting has proven to be an important technique in the analysis of receptor-ligand interactions (i.e., by ligand blotting) and for identifying molecules mediating cell attachment (i.e., by cell blotting). Conventional ligand blotting and cell blotting methods employ non-dynamic (static) incubation conditions, whereby molecules or cells of interest are placed in suspension and overlaid on membranes. However, many cell-cell and cell-matrix adhesive interactions occur under fluid shear conditions, and shear stress itself mediates and/or facilitates the engagement of these physiologically appropriate receptors and ligands. Notably, shear forces critically influence the adhesion of circulating cells and platelets to vessel walls in physiologic cell migration and hemostasis, as well as in inflammatory and thrombotic disorders, cancer metastasis, and atherosclerosis. Use of non-dynamic blotting conditions to analyze such interactions can introduce bias, overtly missing relevant effectors and/or exaggerating the relative role(s) of non-physiologic adhesion molecules. To address this shortfall, we have developed a new technique for identifying binding interactions under fluid shear conditions, the "blot rolling assay." Using this method, molecules in a complex mixture are resolved by gel electrophoresis, transferred to a membrane that is rendered semitransparent, and the membrane is then incorporated into a parallel-plate flow chamber apparatus. Under controlled flow conditions, cells or particles bearing adhesion proteins of interest are then introduced into the chamber and interactions with individual immobilized molecules (bands) can be visualized in real time. The substrate molecule(s) supporting adhesion under fluid shear can then be identified by staining with specific antibodies or by excising the relevant band(s) and performing mass spectrometry or microsequencing of the isolated material. This method thus allows for the identification, within a complex

  7. Western blot analysis of adhesive interactions under fluid shear conditions: the blot rolling assay.

    PubMed

    Sackstein, Robert; Fuhlbrigge, Robert

    2009-01-01

    Western blotting has proven to be an important technique in analysis of receptor-ligand interactions (i.e., by ligand blotting) and for identifying molecules mediating cell attachment (i.e., by cell blotting). Conventional ligand blotting and cell blotting methods employ nondynamic (static) incubation conditions, whereby molecules or cells of interest are placed in suspension and overlaid on membranes. However, many cell-cell and cell-matrix adhesive interactions occur under fluid shear conditions, and shear stress itself mediates and/or facilitates the engagement of these physiologically appropriate receptors and ligands. Notably, shear forces critically influence the adhesion of circulating cells and platelets to vessel walls in physiologic cell migration and hemostasis, as well as in inflammatory and thrombotic disorders, cancer metastasis, and atherosclerosis. Use of nondynamic blotting conditions to analyze such interactions can introduce bias, overtly missing relevant effectors and/or exaggerating the relative role(s) of nonphysiologic adhesion molecules. To address this shortfall, we have developed a new technique for identifying binding interactions under fluid shear conditions, the "blot rolling assay." Using this method, molecules in a complex mixture are resolved by gel electrophoresis, transferred to a membrane that is rendered semi-transparent, and the membrane is then incorporated into a parallel-plate flow chamber apparatus. Under controlled flow conditions, cells or particles bearing adhesion proteins of interest are then introduced into the chamber and interactions with individual immobilized molecules (bands) can be visualized in real-time. The substrate molecule(s) supporting adhesion under fluid shear can then be identified by staining with specific antibodies or by excising the relevant band(s) and performing mass spectrometry or microsequencing of the isolated material. This method thus allows for the identification, within a complex mixture

  8. 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.

  9. Evaluating strategies to normalise biological replicates of Western blot data.

    PubMed

    Degasperi, Andrea; Birtwistle, Marc R; Volinsky, Natalia; Rauch, Jens; Kolch, Walter; Kholodenko, Boris N

    2014-01-01

    Western blot data are widely used in quantitative applications such as statistical testing and mathematical modelling. To ensure accurate quantitation and comparability between experiments, Western blot replicates must be normalised, but it is unclear how the available methods affect statistical properties of the data. Here we evaluate three commonly used normalisation strategies: (i) by fixed normalisation point or control; (ii) by sum of all data points in a replicate; and (iii) by optimal alignment of the replicates. We consider how these different strategies affect the coefficient of variation (CV) and the results of hypothesis testing with the normalised data. Normalisation by fixed point tends to increase the mean CV of normalised data in a manner that naturally depends on the choice of the normalisation point. Thus, in the context of hypothesis testing, normalisation by fixed point reduces false positives and increases false negatives. Analysis of published experimental data shows that choosing normalisation points with low quantified intensities results in a high normalised data CV and should thus be avoided. Normalisation by sum or by optimal alignment redistributes the raw data uncertainty in a mean-dependent manner, reducing the CV of high intensity points and increasing the CV of low intensity points. This causes the effect of normalisations by sum or optimal alignment on hypothesis testing to depend on the mean of the data tested; for high intensity points, false positives are increased and false negatives are decreased, while for low intensity points, false positives are decreased and false negatives are increased. These results will aid users of Western blotting to choose a suitable normalisation strategy and also understand the implications of this normalisation for subsequent hypothesis testing.

  10. Evaluating Strategies to Normalise Biological Replicates of Western Blot Data

    PubMed Central

    Degasperi, Andrea; Birtwistle, Marc R.; Volinsky, Natalia; Rauch, Jens; Kolch, Walter; Kholodenko, Boris N.

    2014-01-01

    Western blot data are widely used in quantitative applications such as statistical testing and mathematical modelling. To ensure accurate quantitation and comparability between experiments, Western blot replicates must be normalised, but it is unclear how the available methods affect statistical properties of the data. Here we evaluate three commonly used normalisation strategies: (i) by fixed normalisation point or control; (ii) by sum of all data points in a replicate; and (iii) by optimal alignment of the replicates. We consider how these different strategies affect the coefficient of variation (CV) and the results of hypothesis testing with the normalised data. Normalisation by fixed point tends to increase the mean CV of normalised data in a manner that naturally depends on the choice of the normalisation point. Thus, in the context of hypothesis testing, normalisation by fixed point reduces false positives and increases false negatives. Analysis of published experimental data shows that choosing normalisation points with low quantified intensities results in a high normalised data CV and should thus be avoided. Normalisation by sum or by optimal alignment redistributes the raw data uncertainty in a mean-dependent manner, reducing the CV of high intensity points and increasing the CV of low intensity points. This causes the effect of normalisations by sum or optimal alignment on hypothesis testing to depend on the mean of the data tested; for high intensity points, false positives are increased and false negatives are decreased, while for low intensity points, false positives are decreased and false negatives are increased. These results will aid users of Western blotting to choose a suitable normalisation strategy and also understand the implications of this normalisation for subsequent hypothesis testing. PMID:24475266

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

    NASA Astrophysics Data System (ADS)

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

    2012-03-01

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

  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. 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.

  14. Serological Diagnosis of Paracoccidioidomycosis through a Western Blot Technique

    PubMed Central

    Perenha-Viana, M. C. Z.; Gonzales, I. A. A.; Brockelt, S. R.; Machado, L. N. C.

    2012-01-01

    Paracoccidioidomycosis (PCM) is a serious infectious disease that progresses toward death if untreated. Its confirmatory diagnosis is made by the detection of the fungus Paracoccidioides brasiliensis in a direct mycological examination or by histopathology. However, these techniques are of low sensitivity. Serological tests seem to be more promising. The objective of this study was to test Western blot (WB) analysis using sera from patients suspected of PCM to determine whether it represents a safe and sensitive serological technique for a rapid and effective diagnosis for this disease. Sera from 517 patients were analyzed through WB analysis and double-immunodiffusion (DID) techniques using a crude exoantigen of P. brasiliensis 339. DID gave positive reactions for 140 sera (27%) and WB for 250 sera (48.4%). All sera that had a positive reaction by DID also had a positive result with a 43-kDa glycoprotein by WB analysis. Among the 377 samples that were negative by DID, 29.1% were reactive in WB analysis. For the cutoff dilution used (1:400), a positive reaction was not observed with any of the 102 sera from patients with other diseases in regions where such diseases are endemic and 30 healthy individuals tested as negative controls. These results prove WB analysis to be a sensitive technique and suggest its inclusion among routine laboratory assays as a safe method for PCM diagnosis. PMID:22301695

  15. The line blot assay: problems with titrating first and second antibodies for Western blot and immunohistochemistry assays?

    PubMed

    Rojas-Espinosa, O; Silva-Miranda, M; Wek-Rodriguez, K; Arce-Paredes, P

    2006-01-01

    We describe a technique designed to assess the optimal dilution of primary and secondary antibodies, to be used in Western blot, dot blot, the multi-antigen print immunoassay (MAPIA) and immunohistochemistry assays. The method that we call "line blot" is not an alternative but a practical, complementary tool for the above techniques that assures definitive results are obtained from single assays, so there is no need to repeat the assay. As with most immunoenzymatic assays, the line blot assay is very sensitive, allowing the detection of absolute amounts of antigen as low as 2.5 ng in the 0.5 cm-long segment line (see Results), depending on the strength of the secondary, enzyme-labelled antibody.

  16. A comparison of the Genie and western blot assays in confirmatory testing for HIV-1 antibody.

    PubMed

    Chan, E L; Sidaway, F; Horsman, G B

    1996-03-01

    The Genie HIV-1/2 kit (Sanofi Diagnostics Pasteur, Montreal, Quebec), a synthetic-peptide solid-phase enzyme immunoassay, was evaluated as a confirmatory assay for HIV-1 antibodies in comparison with Western blot (BioRad, Hercules, CA, USA) on 50 stored HIV-1 antibody-positive sera and the 137 sera yielding repeated positive results in the conventional EIA screen out of 13405 fresh patient sera from Saskatchewan in 1993. The stored HIV-1-positive sera were uniformly positive in the Genie test. Of the 137 EIA screen-positive sera, 33 were uniformly positive and 64 were uniformly negative in Genie and Western blot; 36 were Genie-negative and indeterminate by Western blot; and four were Genie indeterminate, of which one was negative and three were indeterminate by Western blot. All HIV-1 Western blot-indeterminate and Genie-interdeterminate sera were negative in radio-immunoprecipitation assay (RIPA) and Western blot for HIV-1 and HIV-2 antibodies performed by a reference laboratory. Genie gave an accurate definitive result for 97% of EIA positive sera compared with 71% for Western blot. There was excellent correlation between Genie, Western blot and RIPA results. However, the Genie assay was faster, less costly and yielded fewer indeterminate results than Western blot in confirmatory testing for HIV-1 antibodies.

  17. 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…

  18. Detection of antibodies to Hypoderma lineatum in cattle by Western blotting with recombinant hypodermin C antigen.

    PubMed

    Boldbaatar, D; Xuan, X; Kimbita, E; Huang, X; Igarashi, I; Byambaa, B; Battsetseg, B; Battur, B; Battsetseg, G; Batsukh, Z; Nagasawa, H; Fujisaki, K; Mikami, T

    2001-08-01

    The cDNA encoding the entire mature hypodermin C (HC) of Hypoderma lineatum was cloned and expressed in Escherichia coli as a glutathione S-transferase fusion protein using pGEX vector. The recombinant HC protein (rHC) was tested by Western blotting to detect antibodies to H. lineatum in cattle. Western blotting with rHC as antigen clearly differentiated between H. lineatum-infested cattle sera and normal cattle sera. Forty-six out of forty-eight serum samples from cattle in Central Mongolia were positive, whereas all 30 serum samples from cows in Hokkaido, Japan, were negative by Western blotting. The result of Western blotting was identical to that of a previously developed enzyme-linked immunosorbent assay. These data demonstrated that Western blotting, with rHC expressed in E. coli, might be a useful method for the diagnosis of cattle hypodermosis.

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

    PubMed

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

    2015-06-01

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

  20. Indeterminate HIV-1 Western Blots: Etiology, Natural History and Psychological Reactions

    DTIC Science & Technology

    1991-09-24

    viral-like syndrome in the month between the IWB (p24 antibody only) and the positive Western blot. Case 2 was a woman with a history of autoimmune...Dupont blot two weeks later and by Epitope blot four weeks later. Case 3 was a homosexual man with a v!ral-like syndrome who had p2 4 and weak gpl60...proteins on Western blot. Case 4 was a homosexual man with a mononucleosis-like syndrome with a borderline ETA (R-value 0.9) and a faint p24 and weak

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

    PubMed

    Hou, Chenlu; Herr, Amy E

    2013-01-07

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

  2. Reversible Ponceau staining as a loading control alternative to actin in Western blots.

    PubMed

    Romero-Calvo, Isabel; Ocón, Borja; Martínez-Moya, Patricia; Suárez, María Dolores; Zarzuelo, Antonio; Martínez-Augustin, Olga; de Medina, Fermín Sánchez

    2010-06-15

    It is becoming standard practice to measure a housekeeping gene, typically actin, in Western blots, as it is the rule in RNA blots. We have applied reversible Ponceau staining to check equal loading of gels and measured actin in parallel under different conditions. Our results show that densitometric analysis is comparable with both techniques. Therefore, routine quantitation of Ponceau staining before antibody probing is validated as an alternative to actin blotting.

  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. Application of SYPRO Ruby- and Flamingo-stained polyacrylamide gels to Western blot analysis.

    PubMed

    Hagiwara, Makoto; Kobayashi, Ken-Ichi; Tadokoro, Tadahiro; Yamamoto, Yuji

    2010-02-15

    Western blots are widely used for analysis of the expression levels of specific proteins. Blotting is conducted after sodium dodecyl sulfate or native polyacrylamide gel electrophoresis without staining the gel. However, when it is necessary to analyze the gel, duplicate polyacrylamide gels (one of which is stained) usually must be prepared, leading to the consumption of precious sample. Thus, we have developed a convenient and efficient Western blot method using a stained gel. This simple modification should be beneficial for the analysis of samples that are limited in quantity and/or samples for which the stained gel serves as the loading control.

  5. A guide to modern quantitative fluorescent western blotting with troubleshooting strategies.

    PubMed

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

    2014-11-20

    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.

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

    PubMed

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

    2012-09-01

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

  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.

  8. Application of SYPRO Ruby- and Flamingo-stained polyacrylamide gels to Western blot analysis.

    PubMed

    Hagiwara, Makoto; Kobayashi, Ken-Ichi; Tadokoro, Tadahiro; Yamamoto, Yuji

    2009-06-15

    Western blot analysis has been a useful method for analysis of expression levels of specific proteins and is conducted after sodium dodecyl sulfate (SDS) or native polyacrylamide gel electrophoresis without staining the gel. However, when it is necessary to analyze the gel, duplicate polyacrylamide gels usually must be prepared, one of which is stained, leading to the consumption of precious sample. Thus, we developed a convenient and efficient Western blotting method using a stained gel. This simple modification should be beneficial for analyzing samples that are limited in quantity and/or samples for which the stained gel serves as the loading control.

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

    PubMed Central

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

    2014-01-01

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

  10. Western Blot Analysis of the Exotoxin Components From Bacillus anthracis Separated by Isoelectric Focusing Gel Electrophoresis

    DTIC Science & Technology

    2004-02-14

    Biochemical and Biophysical Research Communications 317 (2004) 294–300 295 Approved for...Ames isolate was originally identified as Iowa, it has subsequently been identified as Texas. 296 S.F. Little / Biochemical and Biophysical Research Communications 317... Biochemical and Biophysical Research Communications 317 (2004) 294–300 297 Approved for public release. Distribution is unlimited.Western blots

  11. 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...

  12. 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...

  13. 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…

  14. 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…

  15. 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...

  16. 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

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

    PubMed

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

    2015-01-01

    SDS-PAGE and western blotting are two commonly taught protein detection techniques in biochemistry and molecular biology laboratory classrooms. A pitfall associated with incorporating these techniques into the laboratory is the significant wait times that do not allow students to obtain timely results. The waiting associated with SDS-PAGE comes from staining and destaining, whereas with western blotting it is the times required for antibody incubations and the numerous wash steps. This laboratory exercise incorporates 2,2,2-trichloroethanol (TCE) into the SDS-PAGE gel allowing for visualization of migrated proteins in a matter of minutes, saving both the time and chemical waste associated with traditional Coomassie staining. Additionally, TCE staining does not affect protein transfer eliminating the requirement for duplicated gels for total protein and western analyses. Protein transfer can be confirmed immediately without the use of Ponceau S staining. Lastly, this western blot procedure has been further shortened by using an HRP-conjugated primary antibody, which eliminates the secondary antibody incubation and washes, and uses a colorimetric detection to allow for visualization by students without the need for specialized equipment.

  18. Specific Western Blot Bands Are Associated with Initial CD4+ Lymphocyte Counts in Human Immunodeficiency Virus Seroconverters.

    DTIC Science & Technology

    1992-09-30

    SPECIFIC WESTERN BLOT BANDS ARE ASSOCIATED WITH INITIAL CD4+ LYMPHOCYTE COUNTS IN HUMAN IMMUNODEFICIENCY VIRUS SEROCONVERTERS DTI C ELEkCTE AUG...NAVAL MEDICAL RESEARCH AND DEVELOPMENT COMMAND BETHESDA, MARYLAND Specific Western Blot Bands Are Associated With Initial CD4+ Lymphocyte Counts in...and Ms. Susan Yu also provided - quality assurance review of Western blot results. Mr. Jerry Talicurian, Mr. Juan Jurado, Mr. David Morgan, and

  19. Western blot membrane composed of electrospun polyvinylidene fluoride nanofiber membrane and polyethylene terephthalate sheet.

    PubMed

    Cho, Eugene; Kim, Chan; Hwang, Cheol Ho; Chang, Duck Rye; Kook, Joong-Ki

    2013-06-01

    In a previous study, an electrospun polyvinylidene fluoride (PVDF) nanofiber membrane was developed for Western blotting. The membrane exhibited high sensitivity and high binding capacity for the detection of protein bands that was unlike that observed for conventional, microphase separation-based porous PVDF membranes. Nevertheless, the PVDF nanofiber membrane is quite expensive. The objective of this study was to develop an economical Western blot membrane using a hybrid electrospun PVDF nanofiber and polyethylene terephthalate (PET) sheet. The results showed that the detection sensitivity of the 4 gram per square meter (gsm) membrane was similar to those of the electrospun PVDF nanofiber membrane only, and the 7 gsm PVDF nanofiber membranes on a PET sheet and the electrospun PVDF nanofiber membrane. This means the protein detection sensitivity is not proportional to the thickness of the PVDF nanofiber membrane. The 4 gsm PVDF nanofiber membrane on a PET sheet can be used to detect proteins with high sensitivity and economic efficiency.

  20. Mapping of radiolabeled peptides derived from proteolysis of polypeptides bound to nitrocellulose after Western blotting

    SciTech Connect

    Carrey, E.A.; Hardie, D.G.

    1986-11-01

    Sections of nitrocellulose containing bound /sup 32/P-labeled polypeptides were excised from Western blots and exhaustively digested by trypsin in order to analyze the distribution of phosphorylation sites between the products of limited proteolysis of the multifunctional protein CAD. Using the criterion of analytical isoelectric focusing, the /sup 32/P-peptides obtained by this method were found to be similar, although not identical, to peptides obtained by a more conventional digestion of trichloroacetic acid precipitates. Digestion on Western blots is more straightforward than electrophoretic elution of individual gel slices, gives better recoveries than direct digestion of gel slices, and is particularly suitable for peptide mapping of small peptides which bind to nitrocellulose but would diffuse out of polyacrylamide gels during the commonly used fixing and staining procedures.

  1. Western Blot Analysis of the Exotoxins Components from Bacillus anthracis Separated by Isoelectric Focusing Gel Electrophoresis

    DTIC Science & Technology

    2004-07-01

    Biochemical and Biophysical Research Communications 317 (2004) 294–300 BBRC www.elsevier.com/locate/ybbrcWestern blot analysis...pag iden- tified five different point mutations from 26 different S.F. Little / Biochemical and Biophysical Research Communications 317 (2004) 294...identified as Iowa, it has subsequently been identified as Texas. 296 S.F. Little / Biochemical and Biophysical Research Communications 317 (2004)

  2. Serological Differentiation of Murine Typhus and Epidemic Typhus Using Cross-Adsorption and Western Blotting

    PubMed Central

    La Scola, Bernard; Rydkina, Lena; Ndihokubwayo, Jean-Bosco; Vene, Sirkka; Raoult, Didier

    2000-01-01

    Differentiation of murine typhus due to Rickettsia typhi and epidemic typhus due to Rickettsia prowazekii is critical epidemiologically but difficult serologically. Using serological, epidemiological, and clinical criteria, we selected sera from 264 patients with epidemic typhus and from 44 patients with murine typhus among the 29,188 tested sera in our bank. These sera cross-reacted extensively in indirect fluorescent antibody assays (IFAs) against R. typhi and R. prowazekii, as 42% of the sera from patients with epidemic typhus and 34% of the sera from patients with murine typhus exhibited immunoglobulin M (IgM) and/or IgG titers against the homologous antigen (R. prowazekii and R. typhi, respectively) that were more than one dilution higher than those against the heterologous antigen. Serum cross-adsorption studies and Western blotting were performed on sera from 12 selected patients, 5 with murine typhus, 5 with epidemic typhus, and 2 suffering from typhus of undetermined etiology. Differences in IFA titers against R. typhi and R. prowazekii allowed the identification of the etiological agent in 8 of 12 patients. Western blot studies enabled the identification of the etiological agent in six patients. When the results of IFA and Western blot studies were considered in combination, identification of the etiological agent was possible for 10 of 12 patients. Serum cross-adsorption studies enabled the differentiation of the etiological agent in all patients. Our study indicates that when used together, Western blotting and IFA are useful serological tools to differentiate between R. prowazekii and R. typhi exposures. While a cross-adsorption study is the definitive technique to differentiate between infections with these agents, it was necessary in only 2 of 12 cases (16.7%), and the high costs of such a study limit its use. PMID:10882661

  3. 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.

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

    PubMed Central

    2011-01-01

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

  5. Detection of La/SS-B by western blot using nanogold-tagged antibodies and silver enhancement.

    PubMed

    Maier, Shannon; Hubbell, Sherry; Scofield, R Hal

    2009-01-01

    Immunogold staining with silver enhancement is a versatile, sensitive and specific method for immunodetection of diverse protein antigens separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to nitrocellulose or polyvinylidene difluoride membranes. "Next-generation" antibodies tagged with nanogold particles have a wide scope of use including but not limited to immunohistochemistry, western blotting, electron microscopy, fluorescent activated cell sorting procedures, and cell isolation and migration studies. Herein, we describe the use of a nanogold-tagged anti-mouse IgG secondary antibody and silver enhancement methodologies coupled with antigen-specific unlabeled primary antibodies for the detection of the La/SS-B autoantigen by western blotting as a useful alternative to chemiluminescent and enzymatic detection methods.

  6. Design, preparation and use of ligated phosphoproteins: a novel approach to study protein phosphatases by dot blot array, ELISA and Western blot assays.

    PubMed

    Sun, Luo; Ghosh, Inca; Barshevsky, Tanya; Kochinyan, Samvel; Xu, Ming-Qun

    2007-07-01

    The study of substrate specificity of protein phosphatases (PPs) is very challenging since it is difficult to prepare a suitable phosphorylated substrate. Phosphoproteins, phosphorylated by a protein kinase, or chemically synthesized phosphopeptides are commonly used substrates for PPs. Both types of these substrates have their advantages and limitations. Phosphoproteins mimic more closely the physiologically relevant PP substrates, but their preparation is technically demanding. Synthetic phosphopeptides present advantages over proteins because they can be easily produced in large quantity and their amino acid sequence can be designed to contain potential determinants of substrate specificity. However, short peptides are less optimal compared to in vivo PP substrates and often display poor and variable binding to different matrices, resulting in low sensitivity in analysis of PP activity on solid support. In this work we utilize the intein-mediated protein ligation (IPL) technique to generate substrates for PPs, combining the advantages of proteins and synthetic peptides in one molecule. The ligation of a synthetic phosphopeptide to an intein-generated carrier protein (CP) with a one-to-one stoichiometry results in the formation of a ligated phosphoprotein (LPP). Three widely used assays, dot blot array, Western blot and ELISA were employed to study the PP activity on LPP substrates. Dephosphorylation was measured by detection of the remaining phosphorylation, or lack of it, with a phospho-specific antibody. The data show the advantage of LPPs over free peptides in assays on solid supports. LPPs exhibited enhanced binding to the matrices used in the study, which significantly improved sensitivity and consistency of the assays. In addition, saturation of the signal was circumvented by serial dilution of the assay samples. This report describes detailed experimental procedures for preparation of LPP substrates and their use in PP assays based on immobilization on

  7. FANCD2 Western blot as a diagnostic tool for Brazilian patients with Fanconi anemia.

    PubMed

    Pilonetto, D V; Pereira, N F; Bitencourt, M A; Magdalena, N I R; Vieira, E R; Veiga, L B A; Cavalli, I J; Ribeiro, R C; Pasquini, R

    2009-03-01

    Fanconi anemia is a rare hereditary disease showing genetic heterogeneity due to a variety of mutations in genes involved in DNA repair pathways, which may lead to different clinical manifestations. Phenotypic variability makes diagnosis difficult based only on clinical manifestations, therefore laboratory tests are necessary. New advances in molecular pathogenesis of this disease led researchers to develop a diagnostic test based on Western blot for FANCD2. The objective of the present study was to determine the efficacy of this method for the diagnosis of 84 Brazilian patients with Fanconi anemia, all of whom tested positive for the diepoxybutane test, and 98 healthy controls. The FANCD2 monoubiquitinated isoform (FANCDS+/FANCD2L-) was not detected in 77 patients (91.7%). In 2 patients (2.4%), there was an absence of both the monoubiquitinated and the non-ubiquitinated proteins (FANCD2S-/FANCD2L-) and 5 patients (5.9%) had both isoforms (FANCD2S+/FANCD2L+). This last phenotype suggests downstream subtypes or mosaicism. All controls were diepoxybutane negative and were also negative on the FANCD2 Western blot. The Western blot for FANCD2 presented a sensitivity of 94% (79/84) and specificity of 100% (98/98). This method was confirmed as an efficient approach to screen Brazilian patients with deleterious mutations on FANCD2 (FANCD2S-/FANCD2L-) or other upstream genes of the FA/BRCA pathway (FANCDS+/FANCD2L-), to confirm the chromosome breakage test and to classify patients according to the level of FA/BRCA pathway defects. However, patients showing both FANCD2 isoforms (FANCD2S+/FANCD2L+) require additional studies to confirm mutations on downstream Fanconi anemia genes or the presence of mosaicism.

  8. Western Blotting Is an Efficient Tool for Differential Diagnosis of Paracoccidioidomycosis and Pulmonary Tuberculosis

    PubMed Central

    Bertoni, Thâmara Aline; Perenha-Viana, Maysa Cláudia Zolin; Patussi, Eliana Valéria; Cardoso, Rosilene Fressatti

    2012-01-01

    Sputum and sera from 134 patients screened for tuberculosis (TB) were analyzed to investigate TB and paracoccidioidomycosis (PCM). Of these patients, 11 (8.2%) were confirmed to have TB, but six (4.5%) were positive only for PCM. All patients with PCM presented anti-43-kDa-component antibodies in Western blotting (WB) assays, while in the TB-positive patients these antibodies did not appear. This preliminary study suggests WB as a potential tool for differential laboratory diagnosis between TB and PCM. PMID:22971781

  9. High-selectivity cytology via lab-on-a-disc western blotting of individual cells.

    PubMed

    Kim, John J; Sinkala, Elly; Herr, Amy E

    2017-02-28

    Cytology of sparingly available cell samples from both clinical and experimental settings would benefit from high-selectivity protein tools. To minimize cell handling losses in sparse samples, we design a multi-stage assay using a lab-on-a-disc that integrates cell handling and subsequent single-cell western blotting (scWestern). As the two-layer microfluidic device rotates, the induced centrifugal force directs dissociated cells to dams, which in turn localize the cells over microwells. Cells then sediment into the microwells, where the cells are lysed and subjected to scWestern. Taking into account cell losses from loading, centrifugation, and lysis-buffer exchange, our lab-on-a-disc device handles cell samples with as few as 200 cells with 75% cell settling efficiencies. Over 70% of microwells contain single cells after the centrifugation. In addition to cell settling efficiency, cell-size filtration from a mixed population of two cell lines is also realized by tuning the cell time-of-flight during centrifugation (58.4% settling efficiency with 6.4% impurity). Following the upstream cell handling, scWestern analysis detects four proteins (GFP, β-TUB, GAPDH, and STAT3) in a glioblastoma cell line. By integrating the lab-on-a-disc cell preparation and scWestern analysis, our platform measures proteins from sparse cell samples at single-cell resolution.

  10. Nitrotyrosine Density of Rabbit Urinary Bladder Muscle and Mucosa Measured via Western Blotting and 96-Well Plate Analysis.

    PubMed

    Fitzpatrick, Brittany; Schuler, Catherine; Leggett, Robert E; Levin, Robert M

    2012-01-01

    Purpose. Nitrotyrosine was quantitated in rabbit bladder muscle and mucosa using two analytical systems: Western blotting analyses and a 96-well plate quantitative analysis kit. Materials and Methods. Rabbit bladder muscle and mucosa were obtained from control rabbits. For the Western analysis, the samples were loaded into a SDS page gel and then transferred to a PVDF membrane. The optical density was measured using a Kodak Scanner. Using the 96-well plate, the samples and standards were loaded, incubated with primary and secondary antibody, washed and vacuumed with 10x wash buffer three times between each incubation period. Stop buffer was added to the plate and the results were quantified via the plate reader. Results. For both muscle and mucosa tissue, the optical density readings were linear with tissue concentration; the concentration of nitrotyrosine in the mucosa was significantly higher than in the muscle. However, whereas the Western blot analysis is based on relative optical densities, the 96-well plate kit provides a truly quantitative analysis. Discussion. Mucosa tissue displayed a higher density of nitrotyrosine than did detrusor muscle tissue. This may well be due to the significantly higher metabolic activity of the mucosa compared to the muscle.

  11. Mechanism of DNA (Southern) and protein (Western) blotting on cellulose nitrate and other membranes.

    PubMed

    Van Oss, C J; Good, R J; Chaudhury, M K

    1987-03-27

    The transfer of DNA fractions from hydrophilic gels to nitrocellulose membranes (Southern blotting) which was soon followed by the description of an analogous procedure for RNA (Northern blotting), and somewhat later for proteins (Western blotting), has rapidly become an important separation and characterization method in molecular biology, genetic engineering, and immunological detection. Surface tension measurements have shown that the interfacial attraction between DNA and cellulose esters (-delta G132) in aqueous media can be considerable. The weaker binding energy of proteins to cellulose nitrate and to cellulose acetate may be compared to hydrophobic interaction chromatography, as on account of the somewhat lower [-delta G132] values, it often is necessary to "fix" them more tightly onto nitrocellulose by using high salt concentrations. The binding energy of RNA to both cellulose esters also is rather low. In addition to the effect of high ionic strength, the effect of adding methanol, and the effects of denaturation, heating and drying on the energy of attachment of the biopolymers to cellulose esters, have been studied. Cationized nylon membranes have been advocated recently, especially for electrophoretic transfer of nucleic acids (in which process high salt concentrations cannot easily be used). With positively charged nylon membranes, the attachment mainly occurs through the electrostatic attraction between the strongly negatively charged nucleic acids (or proteins) and the positively charged membrane. Also, more apolar membranes (of polyvinyl difluoride) have been proposed, which manifest a strong interfacial (hydrophobic) attraction to all the above biopolymers (regardless of their electrostatic charge). However, with these two novel membrane types it is no longer possible to exploit the large difference in binding energy between DNA and RNA, which makes cellulose nitrate membranes so uniquely suited for RNA-DNA hybridization assays.

  12. Recombinant Dense Granular Protein (GRA5) for Detection of Human Toxoplasmosis by Western Blot

    PubMed Central

    Ching, Xiao Teng; Lau, Yee Ling; Fong, Mun Yik; Nissapatorn, Veeranoot; Andiappan, Hemah

    2014-01-01

    Toxoplasma gondii infects all warm-blooded animals, including humans, causing serious public health problems and great economic loss for the food industry. Commonly used serological tests require costly and hazardous preparation of whole Toxoplasma lysate antigens from tachyzoites. Here, we have evaluated an alternative method for antigen production, which involved a prokaryotic expression system. Specifically, we expressed T. gondii dense granular protein-5 (GRA5) in Escherichia coli and isolated it by affinity purification. The serodiagnostic potential of the purified recombinant GRA5 (rGRA5) was tested through Western blot analysis against 212 human patient serum samples. We found that rGRA5 protein was 100% specific for analysis of toxoplasmosis-negative human sera. Also, rGRA5 was able to detect acute and chronic T. gondii infections (sensitivities of 46.8% and 61.2%, resp.). PMID:24987700

  13. Western blot analysis of Src kinase assays using peptide substrates ligated to a carrier protein.

    PubMed

    Xu, Jie; Sun, Luo; Ghosh, Inca; Xu, Ming-Qun

    2004-06-01

    We have applied intein-mediated peptide ligation (IPL) to the use of peptide substrates for kinase assays and subsequent Western blot analysis. IPL allows for the efficient ligation of a synthetic peptide with an N-terminal cysteine residue to an intein-generated carrier protein containing a cysteine reactive C-terminal thioester through a native peptide bond. A distinct advantage of this procedure is that each carrier protein molecule ligates only one peptide, ensuring that the ligation product forms a sharp band on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). We demonstrate the effectiveness of this approach by mutational analysis of peptide substrates derived from human cyclin-dependent kinase, Cdc2, which contains a phosphorylation site of human c-Src protein tyrosine kinase.

  14. Direct identification and characterization of llama (Lama glama L.) whey proteins by microsequencing after western blotting.

    PubMed

    Cantisani, A; Napolitano, L; Giuffrida, M G; Conti, A

    1990-01-01

    Amino acid sequence determination is the most reliable and powerful tool to identify a protein or to classify a new one by comparison of its primary structure with already known sequences. A rapid and simple purification procedure is an essential pre-requisite for routine sequence determination. Structural characterization of llama whey proteins was undertaken for evolutionary as well as economic purposes. N-terminal sequence analyses directly on an immobilon polyvinylidene difluoride (PVDF) membrane, following Western blotting of both native and SDS-denatured llama whey proteins after polyacrylamide gel electrophoresis, revealed three different forms of glycosylated alpha-lactalbumin, and a protein with a high degree of homology with a camel whey protein of unknown function. Furthermore, by immunoblotting techniques, the electrophoretic band corresponding to serum albumin was identified.

  15. An overview of technical considerations for Western blotting applications to physiological research.

    PubMed

    Bass, J J; Wilkinson, D J; Rankin, D; Phillips, B E; Szewczyk, N J; Smith, K; Atherton, P J

    2017-01-01

    The applications of Western/immunoblotting (WB) techniques have reached multiple layers of the scientific community and are now considered routine procedures in the field of physiology. This is none more so than in relation to skeletal muscle physiology (i.e., resolving the mechanisms underpinning adaptations to exercise). Indeed, the inclusion of WB data is now considered an essential aspect of many such physiological publications to provide mechanistic insight into regulatory processes. Despite this popularity, and due to the ubiquitous and relatively inexpensive availability of WB equipment, the quality of WB in publications and subsequent analysis and interpretation of the data can be variable, perhaps resulting in spurious conclusions. This may be due to poor laboratory technique and/or lack of comprehension of the critical steps involved in WB and what quality control procedures should be in place to ensure robust data generation. The present review aims to provide a detailed description and critique of WB procedures and technicalities, from sample collection through preparation, blotting and detection, to analysis of the data collected. We aim to provide the reader with improved expertise to critically conduct, evaluate, and troubleshoot the WB process, to produce reproducible and reliable blots.

  16. Western blot diagnosis of vivax malaria with multiple stage-specific antigens of the parasite

    PubMed Central

    Son, Eui-Sun; Kim, Tong Soo

    2001-01-01

    Western blot analysis was performed to diagnose vivax malaria using stage-specific recombinant antigens. Genomic DNA from the whole blood of a malaria patient was used as templates to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), merozoite surface protein (MSP-1), apical merozoite antigen (AMA-1), serine repeat antigen (SERA), and exported antigen (EXP-1) of Plasmodium vivax. Each amplified DNA fragment was inserted into a pGEX-4T plasmid to induce the expression of GST fusion protein in Escherichia coli by IPTG. The bacterial cell extracts were separated on 10% SDS-PAGE followed by western blot analysis with patient sera which was confirmed by blood smear examination. When applied with patient sera, 147 (91.9%) out of 160 vivax malaria, 12 (92.3%) out of 13 falciparum malaria, and all 9 vivax/falciparum mixed malaria reacted with at least one antigen, while no reactions occurred with 20 normal uninfected sera. In the case of vivax malaria, CSP-1 reacted with 128 (80.0%) sera, MSP-1 with 102 (63.8%), AMA-1 with 128 (80.0%), SERA with 115 (71.9%), and EXP-1 with 89 (55.6%), respectively. We obtained higher detection rates when using 5 antigens (91.9%) rather than using each antigen solely (55.6-80%), a combination of 2 (76.3-87.5%), 3 (85.6-90.6%), or 4 antigens (89.4-91.3%). This method can be applied to serological diagnosis, mass screening in endemic regions, or safety test in transfusion of prevalent vivax malaria. PMID:11441504

  17. Western blot diagnosis of vivax malaria with multiple stage-specific antigens of the parasite.

    PubMed

    Son, E S; Kim, T S; Nam, H W

    2001-06-01

    Western blot analysis was performed to diagnose vivax malaria using stage-specific recombinant antigens. Genomic DNA from the whole blood of a malaria patient was used as templates to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), merozoite surface protein (MSP-1), apical merozoite antigen (AMA-1), serine repeat antigen (SERA), and exported antigen (EXP-1) of Plasmodium vivax. Each amplified DNA fragment was inserted into a pGEX-4T plasmid to induce the expression of GST fusion protein in Escherichia coli by IPTG. The bacterial cell extracts were separated on 10% SDS-PAGE followed by western blot analysis with patient sera which was confirmed by blood smear examination. When applied with patient sera, 147 (91.9%) out of 160 vivax malaria, 12 (92.3%) out of 13 falciparum malaria, and all 9 vivax/falciparum mixed malaria reacted with at least one antigen, while no reactions occurred with 20 normal uninfected sera. In the case of vivax malaria, CSP-1 reacted with 128 (80.0%) sera, MSP-1 with 102 (63.8%), AMA-1 with 128 (80.0%), SERA with 115 (71.9%), and EXP-1 with 89 (55.6%), respectively. We obtained higher detection rates when using 5 antigens (91.9%) rather than using each antigen solely (55.6-80%), a combination of 2 (76.3-87.5%), 3 (85.6-90.6%), or 4 antigens (89.4-91.3%). This method can be applied to serological diagnosis, mass screening in endemic regions, or safety test in transfusion of prevalent vivax malaria.

  18. 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.

  19. Contribution of a Comparative Western Blot Method to Early Postnatal Diagnosis of Congenital Syphilis

    PubMed Central

    Foschi, Claudio; Capretti, Maria Grazia; Nardini, Paola; Compri, Monica; Corvaglia, Luigi Tommaso; Faldella, Giacomo; Cevenini, Roberto

    2016-01-01

    Serology has a pivotal role in the diagnosis of congenital syphilis (CS), but problems arise because of the passive transfer of IgG antibodies across the placenta. The aim of this study was to assess the diagnostic value of a comparative Western blot (WB) method finalized to match the IgG immunological profiles of mothers and their own babies at birth in order to differentiate between passively transmitted maternal antibodies and antibodies synthesized by the infants against Treponema pallidum. Thirty infants born to mothers with unknown or inadequate treatment for syphilis were entered in a retrospective study, conducted at St. Orsola-Malpighi Hospital, Bologna, Italy. All of the infants underwent clinical, instrumental, and laboratory examinations, including IgM WB testing. For the retrospective study, an IgG WB assay was performed by blotting T. pallidum antigens onto nitrocellulose sheets and incubating the strips with serum specimens from mother-child pairs. CS was diagnosed in 11 out of the 30 enrolled infants; 9/11 cases received the definitive diagnosis within the first week of life, whereas the remaining two were diagnosed later because of increasing serological test titers. The use of the comparative IgG WB testing performed with serum samples from mother-child pairs allowed a correct CS diagnosis in 10/11 cases. The CS diagnosis was improved by a strategy combining comparative IgG WB results with IgM WB results, leading to a sensitivity of 100%. The comparative IgG WB test is thus a welcome addition to the conventional laboratory methods used for CS diagnosis, allowing identification and adequate treatment of infected infants and avoiding unnecessary therapy of uninfected newborns. PMID:26961856

  20. 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.

  1. The prevalence and significance of HTLV-I/II seroindeterminate Western blot patterns.

    PubMed

    Abrams, Anna; Akahata, Yoshimi; Jacobson, Steven

    2011-08-01

    Human T-lymphotropic virus type I (HTLV-I) infects an estimated 15-20 million persons worldwide. A number of diseases have been associated with the virus including adult T-cell leukemia (ATL), HTLV-associated myelopathy/tropical spastic paraparesis (HAM/TSP), HTLV-I uveitis, and HTLV-I-associated infective dermatitis. Once it was shown that there is an increased risk for developing HAM/TSP associated with blood transfusion, screening for HTLV-1 among blood banks was implemented in Japan, United States, France, and the Netherlands. This process includes detection by an enzyme immunoassay (EIA) followed by a confirmatory Western blot (WB) in which recombinant proteins specific for HTLV-I Env glycoproteins are incorporated into WB strips. HTLV-I seropositive results are defined by the presence of antibodies against either gp46 or gp62/68 (both Env protein bands) and either p19, p24, or p53 (one of the gag bands). HTLV-II seropositivity is confirmed by the presence of rgp46-II. However, numerous cases have been documented in which serum samples are reactive by EIA, but an incomplete banding pattern is displayed by subsequent confirmatory WB. Although the significance of these HTLV-I/II seroindeterminates is unclear, it may suggest a much higher incidence of exposure to HTLV-I/II than previously estimated.

  2. Characterization of excretory-secretory antigens of adult Toxocara canis by western blotting.

    PubMed

    Sudhakar, N R; Samanta, S; Sahu, Shivani; Raina, O K; Gupta, S C; Goswami, T K; Lokesh, K M; Kumar, Ashok

    2014-06-01

    Toxocara canis is one of the most common helminth worm of dogs which continues to stimulate both public health concern alongside the higher scientific interest. It may cause visceral and ocular damage in humans especially in children. The identification of specific antigens of T. canis is important so as to develop better diagnostic techniques. Excretory-secretory (ES) antigens were prepared by culturing the adult T. canis worms in RPMI 1640 medium without serum supplementation followed by ammonium sulphate precipitation. These antigens were separated using sodium dodecyl sulphate-electrophoresis (SDS-PAGE). Recovered proteins ranged from 30 to 384 kDa. The specific reactivity of the T. canis excretory-secretory (TC-ES) proteins was checked by western blotting. The immuno-reactivity of the naturally infected dog sera with the TC-ES antigens showed five bands at 43, 57,105, 139 and 175 kDa. The immuno-reactivity of the hyper immune serum raised in rabbits against TC-ES antigens was observed with ten polypeptides of 21, 25, 30, 37, 45, 50, 57, 69, 77 and 105 kDa. Common antigens band were observed at 57 and 105 KDa. These antigens merit further evaluation as candidate for use in diagnosis of toxocariasis in humans and adult dogs.

  3. HIV-1: absence of infection in subjects with indeterminate western blot.

    PubMed

    Migali, E; Mariotti, D; Lovari, A; Tenani, T; Imperiali, P; Ozzola, G

    1993-01-01

    The search for specific antibodies against the Human Immunodeficiency Virus type-1 (Ab anti HIV 1), using an immunoenzymatic test and a subsequent confirmation test (Western Blot-WB) in patients who were previously positive or borderline at the first test, singled out from about 12500 tested subjects, fifteen patients with indeterminate WB (WBi) for the presence of an abnormal band. The presence of p24 was predominant in those WBi (about 50%); generally p24 was the only band found. In all serum samples with WBi, the viral antigen p24 (Ag p24) was absent. In 3 subjects with WBi, after at least six months from the first test, not only did the same pattern persist but also the search for VIH-1 DNA sequence using the polymerase chain reaction (PCR) gave a negative result. According to our experience and the literature on the subject, we suggest that patients with low risk of infection and whose WBi does not modify with time have a remote possibility of being infected by HIV.

  4. Western blot analysis of antibody response to pneumococcal protein antigens in a murine model of pneumonia.

    PubMed Central

    Mouneimne, H; Juvin, M; Beretti, J L; Azoulay-Dupuis, E; Vallee, E; Geslin, P; Petitpretz, P; Berche, P; Gaillard, J L

    1997-01-01

    To detect new antigen candidates for serological tests, we studied the antibody response to pneumococcal protein antigens in mice infected intratracheally with various Streptococcus pneumoniae strains. Sera were tested by Western blotting against whole-cell protein extracts. Mice developed a detectable immunoglobulin G-type response against a small number of polypeptides. The antibody response was strain dependent: sera from individuals infected with the same strain gave similar banding patterns on immunoblots. The banding patterns varied with the strain used for infection. However, a band at 36 to 38 kDa was recognized by all reactive sera. This band appeared to correspond to a polypeptide that was antigenically well conserved among the different S. pneumoniae serotypes. An antibody response to this antigen developed in mice irrespective of the capsular type, the virulence, and the susceptibility to penicillin G of the infecting strain. Thus, this 36- to 38-kDa protein antigen may be of value for the development of a serological test for humans. PMID:9384307

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

    PubMed

    Gilda, Jennifer E; Gomes, Aldrin V

    2013-09-15

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

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

    PubMed Central

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

    2014-01-01

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

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

    PubMed Central

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

    1999-01-01

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

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

    PubMed

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

    2015-01-01

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

  9. Electrostatic protein immobilization using charged polyacrylamide gels and cationic detergent microfluidic Western blotting.

    PubMed

    Kim, Dohyun; Karns, Kelly; Tia, Samuel Q; He, Mei; Herr, Amy E

    2012-03-06

    We report a novel protein immobilization matrix for fully integrated microfluidic Western blotting (WB). The electrostatic immobilization gel (EIG) enables immobilization of all proteins sized using cetyl trimethylammonium bromide polyacrylamide gel electrophoresis (CTAB-PAGE), for subsequent electrophoretic probing with detection affinity reagents (e.g., labeled antibodies). The "pan-analyte" capture strategy introduced here uses polyacrylamide gel grafted with concentrated point charges (zwitterionic macromolecules), in contrast to existing microfluidic WB strategies that rely on a sandwich immunoassay format for analyte immobilization and detection. Sandwich approaches limit analyte immobilization to capture of only a priori known targets. A charge interaction mechanism study supports the hypothesis that electrostatic interaction plays a major role in analyte immobilization on the EIG. We note that protein capture efficiency depends on both the concentration of copolymerized charges and ionic strength of the gel buffer. We demonstrate pan-analyte immobilization of sized CTAB-laden model proteins (protein G, ovalbumin, bovine serum albumin, β-galactosidase, lactoferrin) on the EIG with initial capture efficiencies ranging from 21 to 100%. Target proteins fixed on the EIG (protein G, lactoferrin) are detected using antibody probes with signal-to-noise ratios of 34 to 275. The approach advances protein immunoblotting performance through 200× reduction on sample consumption, 12× reduction in assay duration, and automated assay operation, compared to slab-gel WB. Using the microfluidic WB assay, assessment of lactoferrin in human tear fluid is demonstrated with a goal of advancing toward nonbiopsy-based diagnosis of Sjögren's Syndrome, an autoimmune disease.

  10. Antibody responses to Borrelia burgdorferi detected by western blot vary geographically in Canada

    PubMed Central

    Ogden, Nicholas H.; Arsenault, Julie; Hatchette, Todd F.; Mechai, Samir; Lindsay, L. Robbin

    2017-01-01

    Lyme disease is emerging in eastern and central Canada, and most cases are diagnosed using the two-tier serological test (Enzyme Immuno Assay [EIA] followed by Western blot [WB]). Simplification of this algorithm would be advantageous unless it impacts test performance. In this study, accuracy of individual proteins of the IgG WB algorithm in predicting the overall test result in samples from Canadians was assessed. Because Borrelia burgdorferi strains vary geographically in Canada, geographic variations in serological responses were also explored. Metrics of relative sensitivity, specificity and the kappa statistic measure of concordance were used to assess the capacity of responses to individual proteins to predict the overall IgG WB result of 2524 EIA (C6)-positive samples from across Canada. Geographic and interannual variations in proportions of samples testing positive were explored by logistic regression. No one protein was highly concordant with the IgG WB result. Significant variations were found amongst years and geographic regions in the prevalence of samples testing positive using the overall IgG WB algorithm, and for individual proteins of the algorithm. In most cases the prevalence of samples testing positive were highest in Nova Scotia, and lower in samples from Manitoba westwards. These findings suggest that the current two tier test may not be simplified and continued use of the current two-tier test method and interpretation is recommended. Geographic and interannual variations in the prevalence of samples testing positive may be consistent with B. burgdorferi strain variation in Canada, and further studies are needed to explore this. PMID:28182723

  11. Evaluation of immunodominant proteins of Mycobacterium avium paratuberculosis cell wall by Western blot analysis.

    PubMed

    Hashemi, Maryam; Madani, Rasool; Razmi, Nematollah

    2014-04-01

    Mycobacterium avium subspecies paratuberculosis (M. paratuberculosis) is a slow growing mycobactin, whose dependence on mycobacterial species is known to be the causative agent of Johne's disease (paratuberculosis) in all species of domestic ruminants worldwide. The organism is transmitted via close contact, ingestion, or transplacentally from mother to fetus and occurs commonly in grazing domestic animals. Johne's disease (JD) is characterized by gradual weight loss, decreased milk production, and diarrhea due to the chronic, progressive, granulomatous enteritis and lymphadenitis. The disease can cause serious economic damage to the dairy industry due to the loss of milk production and early culling of infected animals. In recent years, researchers have focused on the identification of a specific antigen of M. paratuberculosis to use in diagnosis test and preparation of effective vaccine. The goal of this study is evaluation of the immunodominant proteins of M. paratuberculosis cell wall. The amount of protein was determined with a Lowry assay (22.68 μg/100 μL). For production of polyclonal antibody against proteins of M. paratuberculosis cell wall, a New Zealand white rabbit was immunized with antigen and Freund's adjuvant. After immunization, the rabbit was bled to produce enriched serum. Antibodies were purified from serum with ion exchange chromatography. In the Ouchterlony test, the reactions between antigen and antibodies were seen in dilutions of one quarter for serum, one quarter for Ig, and one half for IgG by clear precipitation lines due to the well immunization of the rabbit. Electrophoresis and Western blot analysis were used and subsequently a sharp band appeared in nitrocellulose paper; these bands were about 25, 37, 50, 75, and 150 kDa molecular weight, which indicated immunodominant proteins.

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

    PubMed

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

    2015-12-01

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

  13. 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...

  14. Applications of an Automated and Quantitative CE-Based Size and Charge Western Blot for Therapeutic Proteins and Vaccines.

    PubMed

    Rustandi, Richard R; Hamm, Melissa; Lancaster, Catherine; Loughney, John W

    2016-01-01

    Capillary Electrophoresis (CE) is a versatile and indispensable analytical tool that can be applied to characterize proteins. In recent years, labor-intensive SDS-PAGE and IEF slab gels have been replaced with CE-SDS (CGE) and CE-IEF methods, respectively, in the biopharmaceutical industry. These two CE-based methods are now an industry standard and are an expectation of the regulatory agencies for biologics characterization. Another important and traditional slab gel technique is the western blot, which detects proteins using immuno-specific reagents after SDS-PAGE separation. This technique is widely used across industrial and academic laboratories, but it is very laborious, manual, time-consuming, and only semi-quantitative. Here, we describe the applications of a relatively new CE-based western blot technology which is automated, fast, and quantitative. We have used this technology for both charge- and size-based CE westerns to analyze biotherapeutic and vaccine products. The size-based capillary western can be used for fast antibody screening, clone selection, product titer, identity, and degradation while the charge-based capillary western can be used to study product charge heterogeneity. Examples using this technology for monoclonal antibody (mAb), Enbrel, CRM197, and Clostridium difficile (C. difficile) vaccine proteins are presented here to demonstrate the utility of the capillary western techniques. Details of sample preparation and experimental conditions for each capillary western mode are described in this chapter.

  15. Stain-free detection as loading control alternative to Ponceau and housekeeping protein immunodetection in Western blotting.

    PubMed

    Rivero-Gutiérrez, B; Anzola, A; Martínez-Augustin, O; de Medina, F Sánchez

    2014-12-15

    It is currently a routine practice to require a measurement of a housekeeping reference, including actin, glyceraldehyde-3-phosphate dehydrogenase, β-tubulin, among others, in Western blots, as it is the rule in RNA blots. Reversible Ponceau staining has been applied successfully to check equal loading of gels. Here we test a new technique, with the Stain-Free gels from Bio-Rad, against both Ponceau staining and housekeeping protein immunodetection under different conditions. Our results show that Stain-Free gels outperform Ponceau staining and that both are more consistent than housekeeping proteins as a loading control.

  16. Stabilization of hypoxia-inducible factor-1α in buffer containing cobalt chloride for Western blot analysis.

    PubMed

    Srinivasan, Sathyanarayanan; Dunn, Jeff F

    2011-09-01

    Hypoxia-inducible factor-1α (HIF-1α) is a widely studied protein with significant biomedical impact. Care is needed to stabilize HIF-1α protein during sample preparation for Western blot analysis due to its rapid degradation in the presence of oxygen. Enzyme inhibitor cocktails can be complex and expensive. We present a protease inhibitor-free buffer, containing cobalt chloride, which is effective at stabilizing HIF-1α, while being inexpensive, straightforward, and convenient, and has potential for widespread application.

  17. Western blotting of formaldehyde-fixed neuropeptides as small as 400 daltons on gelatin-coated nitrocellulose paper.

    PubMed

    Too, C K; Murphy, P R; Croll, R P

    1994-06-01

    A method is described for Western blotting of peptides as small as 400 daltons (Da). Peptides were separated by tricine-sodium dodecyl sulfate electrophoresis and electroblotted to gelatin-coated PH79 nitrocellulose paper (0.1 micron). The electroblotted peptides were fixed to the nitrocellulose paper for 5-10 min in 4% paraformaldehyde solution. Using anti-rabbit FMRF-amide (Phe-Met-Arg-Phe-NH2) as primary antibody, positive immunoreactivity was detected with an amplified alkaline phosphatase assay which was sensitive to at least 0.5 microgram FMRFamide/lane. When immunoreactivity was determined with 125I-protein A, it was possible to amplify and detect weak signals by increasing the autoradiography time. Therefore, using the 125I-protein A detection method, Western blot analysis of brain extracts from Lymnaea stagnalis (pond snail) and Poecilia reticulata (guppy) indicated the presence of four FMRFamide immunoreactive bands after a 7-day exposure to X-ray film. The most abundant peptide coelectrophoresed with the FMRFamide standard (M(r) 598.8 Da). In addition, this Western blotting procedure also detected APGWamide (Ala-Pro-Gly-Try-NH2; 428.5 Da) and [D-Ala2]-Leu-enkephalinamide (568.7 Da) with their respective specific antibodies.

  18. Differential detection of cytoplasmic Wilms tumor 1 expression by immunohistochemistry, western blotting and mRNA quantification.

    PubMed

    Maki, Takehiro; Ikeda, Hiroaki; Kuroda, Aki; Kyogoku, Noriaki; Yamamura, Yoshiyuki; Tabata, Yukiko; Abiko, Takehiro; Tsuchikawa, Takahiro; Hida, Yasuhiro; Shichinohe, Toshiaki; Tanaka, Eiichi; Kaga, Kichizo; Hatanaka, Kanako; Matsuno, Yoshihiro; Imai, Naoko; Hirano, Satoshi

    2017-01-01

    Wilms tumor 1 (WT1) is considered to be a promising target of cancer treatment because it has been reported to be frequently expressed at high levels in various malignancies. Although WT1-targeted cancer treatment has been initiated, conclusive detection methods for WT1 are not established. The present study aimed to consolidate immunohistochemistry for WT1 with statistical basis. Transfected cells with forced WT1 expression yielded specific western blot bands and nuclear immunostaining; cytoplasmic immunostaining was not specifically recognized. Immunohistochemistry, western blotting, and quantitative reverse transcriptase-polymerase chain reaction were performed in 35 human cell lines using multiple WT1 antibodies and their results were quantified. Relationships among the quantified results were statistically analyzed; the nuclear immunostaining positively correlated with western blot bands and mRNA expression levels, whereas cytoplasmic immunostaining did not. These results indicate that nuclear immunostaining reflects WT1 expression but cytoplasmic immunostaining does not. The nuclear immunostaining was barely (3/541) observed in primary cancer of esophagus, bile duct, pancreas and lung. Although the present study has some limitations, the results indicate that the cytoplasmic immunostaining does not correlate with actual WT1 expression and prompts researchers to carefully evaluate target molecule expression in treatment of cancer.

  19. Investigation of Anti-Toxocara Antibodies in Epileptic Patients and Comparison of Two Methods: ELISA and Western Blotting

    PubMed Central

    Zibaei, Mohammad; Firoozeh, Farzaneh; Bahrami, Parviz; Sadjjadi, Seyed Mahmoud

    2013-01-01

    The relationship between Toxocara infection and epilepsy was previously demonstrated by several case-control studies and case reports. These previous studies were often based on the enzyme-linked immunosorbent assay (ELISA) using Toxocara excretory-secretory antigens, which are not specific due to cross-reactivity with other parasitic infections such as ascariasis, trichuriasis, and anisakiasis. An immunoblot analysis is highly specific and can detect low levels of Toxocara antibodies. Therefore, this assay may be useful in the identification of toxocariasis in epileptic patients. We examined patients who had epilepsy and healthy subjects for seropositivity for Toxocara infection by ELISA and Western blotting. Out of 85 epileptic patients, 10 (11.8%) and 3 (3.5%) persons exhibited Toxocara immunoglobulin G (IgG) antibodies responses by ELISA and by both techniques, respectively. Moreover, in the healthy group (n = 85), 3 (3.5%) persons were positive by ELISA, but none was detected by Western blotting. This study indicates that Toxocara infection is a risk factor for epilepsy in Iran. These findings strongly suggest the need to perform Western blotting immunodiagnosis, as well as the ELISA using Toxocara excretory-secretory antigens, to improve diagnosis of human toxocariasis in patients with epilepsy. PMID:23710354

  20. Western blot analysis of virus-specific antibody responses for capripox and contagious pustular dermatitis viral infections in sheep.

    PubMed

    Chand, P; Kitching, R P; Black, D N

    1994-10-01

    This paper reports the development and evaluation of serological tests for the differentiation of antibodies in animals infected with capripox and parapox viruses. Agar-gel immunodiffusion tests using sera from sheep with naturally-acquired infections and from sheep experimentally inoculated with orf or capripox viruses showed cross reactions. Virus-specific antibody responses to structural proteins of the viruses were analysed by Western-blot analysis. This analysis readily differentiated the infections as either capripox or contagious pustular dermatitis. The antibody responses to the 32 kDa and 26 kDa proteins of capripoxvirus provided a firm basis for differentiation.

  1. Application of Western blot analysis for the diagnosis of Encephalitozoon cuniculi infection in rabbits: example of a quantitative approach.

    PubMed

    Desoubeaux, Guillaume; Pantin, Ana; Peschke, Roman; Joachim, Anja; Cray, Carolyn

    2017-02-01

    Diagnosis of Encephalitozoon cuniculi infection in rabbits remains a major veterinary issue. ELISA or immunofluorescence assays are the current reference standards of serological tests. However, these conventional techniques suffer from a lack of accuracy for distinguishing active from past infections, as a positive serostatus is common in clinically normal rabbits. In this study, we assessed the diagnostic performance of Western blot (WB) to detect both anti-E. cuniculi immunoglobulin G (IgG) and immunoglobulin M (IgM) in comparison with ELISA and to address the intensity of the immune response through a quantitative approach. Positive WB results were highly correlated with the E. cuniculi-related diseased status (P < 0.0001). Although it was more labor intensive and less standardized, quantitative WB provided detailed comparable analysis regarding the humoral response and diagnostic performance similar to ELISA testing with statistically higher sensitivity (88.4 vs. 76.1% for IgG detection and 84.3 vs. 70.4% for IgM, P < 0.01). Several specific WB bands were shown to be significantly associated with concomitant clinical signs, like the one located at 50 kDa (OR = 8.2, [2.4-27.7], P = 0.0008) for IgG and (OR = 27.9, [4.2-187.9], P = 0.0006) for IgM. Therefore, the quantitative WB may have application in veterinary diagnostic laboratories to increase the accuracy of the clinical diagnosis of E. cuniculi infection. In addition, this tool may help to further understand the development and function of the humoral immune response to this infectious agent.

  2. Quantum dot bio-conjugate: as a western blot probe for highly sensitive detection of cellular proteins

    NASA Astrophysics Data System (ADS)

    Kale, Sonia; Kale, Anup; Gholap, Haribhau; Rana, Abhimanyu; Desai, Rama; Banpurkar, Arun; Ogale, Satishchandra; Shastry, Padma

    2012-03-01

    In the present study, we report a quantum dot (QD)-tailored western blot analysis for a sensitive, rapid and flexible detection of the nuclear and cytoplasmic proteins. Highly luminescent CdTe and (CdTe)ZnS QDs are synthesized by aqueous method. High resolution transmission electron microscopy, Raman spectroscopy, fourier transform infrared spectroscopy, fluorescence spectroscopy and X-ray diffraction are used to characterize the properties of the quantum dots. The QDs are functionalized with antibodies of prostate apoptosis response-4 (Par-4), poly(ADP-ribose) polymerases and β actin to specifically bind with the proteins localized in the nucleus and cytoplasm of the cells, respectively. The QD-conjugated antibodies are used to overcome the limitations of conventional western blot technique. The sensitivity and rapidity of protein detection in QD-based approach is very high, with detection limits up to 10 pg of protein. In addition, these labels provide the capability of enhanced identification and localization of marker proteins in intact cells by confocal laser scanning microscopy.

  3. 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.

  4. Laboratory diagnosis of hymenoptera venom allergy: comparative study between specific IgE, western blot and allergen leukocyte stimulation (CAST).

    PubMed

    Santos, M C Pereira; Carlos, M L Palma; Pedro, E; Carlos, A G Palma

    2002-01-01

    Allergy to hymenoptera venom is a classical IgE mediated disease with a potentially fatal course. Specific venom immunotherapy (SIT) is the most effective mean of treating this serious condition, after the diagnosis has been clearly established by a clinical history, in-vivo and in vitro tests. We have evaluated the usefulness of a cellular test (CAST) which is a recently developed ELISA method based on the evaluation of sulfidoleukotriene secretion by leukocytes stimulated with specific antigen. We also evaluated the correlation of CAST with skin tests, specific IgE (sIgE) and western blot for sIgE for hymenoptera venom sIgE. We have included in this study 14 patients, with a clinical history suggestive of hymenoptera venom allergy. None of them had previously been subjected to immunotherapy. A good correlation was obtained between skin tests, sIgE and western blot. However, there was no correlation between these methods and CAST. We conclude that the positivity of CAST method raises some questions about other mechanisms, which maybe non-IgE dependent. Although the number of patients in this study is quite small, the immunoblot analysis may be a valuable additional method in insect venom allergy.

  5. 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

  6. Western blot analysis of virus-specific antibody responses for capripox and contagious pustular dermatitis viral infections in sheep.

    PubMed Central

    Chand, P.; Kitching, R. P.; Black, D. N.

    1994-01-01

    This paper reports the development and evaluation of serological tests for the differentiation of antibodies in animals infected with capripox and parapox viruses. Agar-gel immunodiffusion tests using sera from sheep with naturally-acquired infections and from sheep experimentally inoculated with orf or capripox viruses showed cross reactions. Virus-specific antibody responses to structural proteins of the viruses were analysed by Western-blot analysis. This analysis readily differentiated the infections as either capripox or contagious pustular dermatitis. The antibody responses to the 32 kDa and 26 kDa proteins of capripoxvirus provided a firm basis for differentiation. Images Fig. 1 Fig. 2 Fig. 3 Fig. 4 PMID:7925674

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

    PubMed

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

    2015-12-01

    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.

  8. Standardisation of Western blotting to detect HTLV-1 antibodies synthesised in the central nervous system of HAM/TSP patients.

    PubMed

    Ribeiro, Luiz Claudio Pereira; Gonçalves, Cassia Cristina Alves; Slater, Carla Maria Sena Andrade; Carvalho, Silvia Maia Farias de; Puccioni-Sohler, Marzia

    2013-09-01

    Intrathecal synthesis of human T-lymphotropic virus type 1 (HTLV-1) antibodies (Abs) represents conclusive evidence of a specific immune response in the central nervous system of HTLV-1 associated myelopathy/tropical spastic paraparesis (HAM/TSP) patients. Western blotting (WB) for HTLV Abs in serum is a confirmatory test for HTLV-1 infection. The aim of this study was to standardise the Western blot to demonstrate the intrathecal pattern of Abs against HTLV-1 proteins in HAM/TSP patients. Paired cerebrospinal fluid (CSF) and serum samples were selected from 20 patients with definite HAM/TSP, 19 HTLV-1 seronegative patients and two HTLV-1 patients without definite HAM/TSP. The presence of reactive bands of greater intensity in the CSF compared to serum (or bands in only the CSF) indicated the intrathecal synthesis of anti-HTLV-1 Abs. All definite HAM/TSP patients presented with an intrathecal synthesis of anti-HTLV-1 Abs; these Abs were not detected in the control patients. The most frequent intrathecal targets of anti-HTLV-1 Abs were GD21, rgp46-I and p24 and, to a lesser extent, p19, p26, p28, p32, p36, p53 gp21 and gp46. The intrathecal immune response against env (GD21 and rgp46-I) and gag (p24) proteins represents the most important humoral pattern in HAM/TSP. This response may be used as a diagnostic marker, considering the frequent association of intrathecal anti-HTLV-1 Ab synthesis with HAM/TSP and the pathogenesis of this neurological disease.

  9. [Optimization of the immunoelectrophoresis technic (western blot) for the confirmation of human immunodeficiency virus infection (HIV) in Panama].

    PubMed

    Pascale, J M; de Austin, E; de Moreno, N O; Ledezma, C; de Márquez, E; Blanco, R; Quiroz, E; Calzada, J; Vincensini, A R; de Martin, M C

    1995-01-01

    The purpose of this study is to report the results of the authors' investigation to apply the western blot technique (WB UP-LCS) in the diagnosis of human immunodeficiency virus type 1 (HIV-1) infection. To do this, the authors separated the proteins of the HIV-1 virus by electrophoresis, based on their molecular weight, in poliacilamide gel with SDS (SDS-PAGE) during 3 hours at 200 volts. Then they electrotransferred these proteins to nitrocellulose paper during four hours at 200 milliamperes, with the aid of external cooling. The nitrocellulose strips were evaluated considering the incubation time (1 and 16 hours), two conjugates (human anti IgG with Peroxidase and human anti IgG Biotin plus Streptatividine with Peroxidase) and two dilutions of the patients' sera (1/50 and 1/100). Based on their results the Authors conclude that, in the first place, the optimal conditions for the test include a dilution of 1/100 of the patients serum, incubation of the serum for 16 hours and the use of the conjugate of anti human IgG with Biotin and Streptavidine with Peroxidase; secondary, that the immunologic reactivity against proteins p24 and gp 160/120 is the most important diagnostic criterion for the confirmation of infection with HIV-1 and that they obtained a diagnostic correlation of 100% at a cost which was 5 to 7 times less than that of the commercial system.

  10. 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

  11. Analysis of cell wall extracts of Candida albicans by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot techniques.

    PubMed Central

    Ponton, J; Jones, J M

    1986-01-01

    Cell walls of intact yeast- and mycelial-phase Candida albicans B311 were extracted with different compounds: dithiothreitol, dithiothreitol with protease, dithiothreitol with lyticase, and dithiothreitol with protease followed by beta-glucuronidase with chitinase. Extracts were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot techniques. Dithiothreitol extracts contained the most satisfactory array of components for study. Analysis of these extracts demonstrated that the outer cell wall layers of Candida blastoconidia and germ tubes contained a complex array of polysaccharides, glycoproteins, and proteins. The proteins contributed to a latticework stabilized by covalent bonds that was important in determining the porosity of the outer cell wall layers. When equivalent weights were analyzed, mycelial-phase extract contained a more varied array of proteins than did yeast-phase extract. Only a portion of proteins in mycelial-phase extract elicited antibody responses in hyperimmunized rabbits or infected humans. A polysaccharide-rich, high-molecular-weight component (migrating at a position that would correspond to proteins having molecular weights of 235,000 to 250,000) and a protein component (molecular weight, 19,000) were readily demonstrable in the mycelial-phase extract but could not be identified in the yeast-phase extract. Images PMID:3527986

  12. A cost effective non-commercial ECL-solution for Western blot detections yielding strong signals and low background.

    PubMed

    Haan, Claude; Behrmann, Iris

    2007-01-10

    We compared several alternative ECL solutions for Western blot detection of endogenous proteins in whole cell lysates using inexpensive, commercially available reagents. Starting from an existing protocol based on p-coumaric acid (pCA) as enhancer, we found that the ECL solution containing 4-iodophenylboronic acid (4IPBA) generated strong specific signals and low background chemiluminescence. We optimised the luminol, 4IPBA and hydrogenperoxide concentrations of this 4IPBA-ECL solution. The optimised 4IPBA-ECL solution (100 mM Tris/HCl pH 8.8, 1.25 mM luminol, 2 mM 4IPBA, 5.3 mM hydrogenperoxide) shows a greatly increased signal intensity compared to the initial pCA-ECL protocol and to some commercially available ECL solutions. In addition, the optimised 4IPBA-ECL solution also generates much lower background chemiluminescence than other non-commercial ECL solutions using p-coumaric acid or 4-iodophenol as enhancers. The 4IPBA-ECL solution was stable when stored but had the lowest background when prepared freshly from stock solutions. Thus, we present an optimised protocol for a well-performing inexpensive ECL solution which is an alternative to expensive commercial ECL solutions and which achieves a better signal and lower background than the commercial solutions tested.

  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.

  14. 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.

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

    PubMed

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

    2012-04-30

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

  16. 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.

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

    PubMed

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

    2015-12-15

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

  18. 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)

  19. 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...

  20. Human T lymphotropic virus types I and II proviral sequences in Argentinian blood donors with indeterminate Western blot patterns.

    PubMed

    Mangano, A M; Remesar, M; del Pozo, A; Sen, Luisa

    2004-10-01

    Human T-cell lymphotropic virus (HTLV) seroindeterminate blood donors have been reported worldwide including Argentina. To investigate the significance of HTLV-I/II seroindeterminate Western blot (WB) patterns, we conducted an 8-year cross-sectional study. Of 86,238 Argentinian blood donors, 146 sera were reactive by screening tests. The WB results indicated that 20% were HTLV-I reactive, 8% HTLV-II reactive, 61% indeterminate, and 11% negative. The overall seroprevalence was 0.034% for HTLV-I, 0.014% for HTLV-II, and 0.103% for indeterminate. In 57 reactive specimens, HTLV-I/II provirus could be examined by type specific PCR for tax, pol, and env regions. When at least two gene fragments were amplified HTLV-I/II infection was considered confirmed. PCR results confirmed all WB seropositive samples for HTLV-I (n = 15), and HTLV-II (n = 7), and the only WB negative case was also PCR negative, showing a complete concordance between PCR and WB. However, of 34 WB seroindeterminate sera studied by PCR, in 5 was proviral DNA amplified. According to our criteria PCR confirmed one to be HTLV-I, and one HTLV-II, 3 remained indeterminate since only tax sequences were amplified. Among WB indeterminate samples tested by PCR, most of their serological profile showed reactivity to gag codified proteins but lacked env reactivities (70%). One sample with a WB gag pattern showed proviral tax sequences, but of the four samples with reactivity to env proteins GD21 (n = 3) or rgp46II (n = 1) PCR results indicated that one was HTLV-I, one was HTLV-II, and two were indeterminate (only tax sequences). In conclusion, the majority of HTLV-seroindeterminate WB donors exhibited a gag indeterminate profile lacking HTLV provirus, and were thus considered uninfected. However, seroreactivity to env proteins, in particular to GD21, may indicate infection and a follow-up study of each seroreactive blood donor should be considered.

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

    PubMed

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

    2013-12-01

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

  2. A Western blot-based investigation of the yeast secretory pathway designed for an intermediate-level undergraduate cell biology laboratory.

    PubMed

    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 two distinct steps of protein secretion were differentiated using a genetic reporter designed specifically to identify defects in the first step of the pathway, the insertion of proteins into the endoplasmic reticulum (Vallen, 2002). We have developed two versions of a Western blotting assay that serves as a second way of distinguishing the two secretory mutants, which we pair with the genetic assay in a 3-wk laboratory module. A quiz administered before and after students participated in the lab activities revealed significant postlab gains in their understanding of the secretory pathway and experimental techniques used to study it. A second survey administered at the end of the lab module assessed student perceptions of the efficacy of the lab activities; the results of this survey indicated that the experiments were successful in meeting a set of educational goals defined by the instructor.

  3. Avoiding Pitfalls of Internal Controls: Validation of Reference Genes for Analysis by qRT-PCR and Western Blot throughout Rat Retinal Development

    PubMed Central

    Rocha-Martins, Maurício; Njaine, Brian; Silveira, Mariana S.

    2012-01-01

    Background Housekeeping genes have been commonly used as reference to normalize gene expression and protein content data because of its presumed constitutive expression. In this paper, we challenge the consensual idea that housekeeping genes are reliable controls for expression studies in the retina through the investigation of a panel of reference genes potentially suitable for analysis of different stages of retinal development. Methodology/Principal Findings We applied statistical tools on combinations of retinal developmental stages to assess the most stable internal controls for quantitative RT-PCR (qRT-PCR). The stability of expression of seven putative reference genes (Actb, B2m, Gapdh, Hprt1, Mapk1, Ppia and Rn18s) was analyzed using geNorm, BestKeeper and Normfinder software. In addition, several housekeeping genes were tested as loading controls for Western blot in the same sample panel, using Image J. Overall, for qRT-PCR the combination of Gapdh and Mapk1 showed the highest stability for most experimental sets. Actb was downregulated in more mature stages, while Rn18s and Hprt1 showed the highest variability. We normalized the expression of cyclin D1 using various reference genes and demonstrated that spurious results may result from blind selection of internal controls. For Western blot significant variation could be seen among four putative internal controls (β-actin, cyclophilin b, α-tubulin and lamin A/C), while MAPK1 was stably expressed. Conclusion Putative housekeeping genes exhibit significant variation in both mRNA and protein content during retinal development. Our results showed that distinct combinations of internal controls fit for each experimental set in the case of qRT-PCR and that MAPK1 is a reliable loading control for Western blot. The results indicate that biased study outcomes may follow the use of reference genes without prior validation for qRT-PCR and Western blot. PMID:22916200

  4. 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

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

    PubMed

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

    2015-05-01

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

  6. Assessment and modification of a Western blot assay for detection of central nervous system tissue in meat products in the United States.

    PubMed

    Salman, M D; Jemmi, T; Triantis, J; Dewell, R D

    2005-08-01

    Health hazards associated with meat contaminated by the bovine spongiform encephalopathy agent have led to the development of tests for the presence of this agent. The objective of this study was to optimize a neuron-specific enolase Western blot assay for use in the United States. We compared the original test with a modified protocol to evaluate the detection limit for the presence of central nervous system (CNS) tissue in experimentally inoculated samples and compared and evaluated the utility of these tests for detecting CNS tissue in retail sausages. Sensitivity and specificity of the original and modified protocols were evaluated using the kappa statistic to assess agreement between the results of the two protocols. The original protocol resulted in 100% specificity and 92% sensitivity for raw samples and 92% specificity and 72% sensitivity for cooked samples. The modified protocol resulted in 92% specificity and 89% sensitivity for raw samples and 83% specificity and 75% sensitivity for cooked samples. The kappa statistic for protocol comparison was 0.94 for raw samples and 0.74 for cooked samples. Both protocols correctly identified CNS tissue in positive controls for each replicate. Although the Western blot technique should be considered for screening for the presence of bovine CNS tissue in meat samples, the techniques should be further optimized to address problems of low sensitivity. A test with higher sensitivity is needed to protect consumers from food safety threats associated with bovine CNS tissue.

  7. Characterization of antigenic property of Toxocara canis and Toxascaris leonina adults and larvae through immunodiagnostic electrophoresis (SDS-PAGE) and western blot technique.

    PubMed

    el-Massry, A A

    1999-08-01

    Differential molecular studies were performed by sodium dodecyle sulphate polyacrylamide gel electrophoresis--SDS-PAGE--between somatic antigen of Toxocara canis and Toxascaris leonina, adults and larvae, recovered from dogs. SDS-PAGE of both adults somatic antigen showed two closely similar bands (90.00, 91.95 KDa and 69.25-70.56 KDa). Each parasite had characteristic bands clustered in different molecular weights. While for their larval antigen, T. canis showed a very different antigenic profile when analysed in comparison to T. leonina antigen except at one band (66.85-66.89 KDa). The Western blot analysis showed four prominent bands represented immunoreaction between the separated somatic antigen of T. canis adults and experimentally immunized rabbit with the corresponding parasite (125.37, 117.73, 90.00 and 69.25 KDa). While separated antigen of T. leonina adults immune reacted with the corresponding hyperimmune rabbit sera at 119.04, 91.95 and 70.56 KDa. The Western blot showed cross reactive immune bands between T. canis and T. leonina adults somatic antigen at two bands (90.00, 91.95 KDa and 69.25-70.56 KDa). The polypeptide bands reacted at 125.37 KDa and 117.73 KDa can be used as specific finger print for T. canis adults while that at 119.04 KDa was specific for T. leonina adult worm.

  8. Product-selective blot: a technique for measuring enzyme activities in large numbers of samples and in native electrophoresis gels

    SciTech Connect

    Thompson, G.A.; Davies, H.M.; McDonald, N.

    1985-08-01

    A method termed product-selective blotting has been developed for screening large numbers of samples for enzyme activity. The technique is particularly well suited to detection of enzymes in native electrophoresis gels. The principle of the method was demonstrated by blotting samples from glutaminase or glutamate synthase reactions into an agarose gel embedded with ion-exchange resin under conditions favoring binding of product (glutamate) over substrates and other substances in the reaction mixture. After washes to remove these unbound substances, the product was measured using either fluorometric staining or radiometric techniques. Glutaminase activity in native electrophoresis gels was visualized by a related procedure in which substrates and products from reactions run in the electrophoresis gel were blotted directly into a resin-containing image gel. Considering the selective-binding materials available for use in the image gel, along with the possible detection systems, this method has potentially broad application.

  9. Qualitative and quantitative analyses of Epstein-Barr virus early antigen diffuse component by western blotting enzyme-linked immunosorbent assay with a monoclonal antibody.

    PubMed Central

    Lin, J C; Choi, E I; Pagano, J S

    1985-01-01

    We report the use of monoclonal antibody against the early antigen diffuse component (anti-EA-D) of Epstein-Barr virus (EBV) to analyze, both qualitatively and quantitatively, the expression of EA-D in various human lymphoblastoid cell lines activated by chemical inducers. The kinetics of synthesis of EA-D in P3HR-1, B95-8, and Ramos/AW cells were similar in that they all reached the peak of synthesis on day 5 after induction. Surprisingly, no expression of EA-D was found in induced BJAB/GC, an EBV-genome-containing cell line. EBV-negative cell lines, BJAB and Ramos, were negative for EA-D. Raji cells had no detectable EA-D but responded rapidly to induction, reaching a peak on day 3. Superinfection of Raji cells also resulted in marked induction of EA-D, which reached a plateau between 8 to 12 h postinfection. Western blotting coupled with the enzyme-linked immunosorbent assay was employed to identify polypeptides representing EA-D. A family of four polypeptides with molecular weights of 46,000 (46K protein), 49,000, 52,000, and 55,000 were identified to be reactive with monoclonal anti-EA-D antiserum. The pattern of EA-D polypeptides expressed in each cell line was different. Of particular interest was the expression of a large quantity of 46K protein both in induced Raji and P3HR-1 cells, but not in superinfected Raji cells. A 49K doublet was expressed in activated p3HR-1, B95-8, and Ramos/AW cells and in superinfected Raji cells. In addition, two distinct 52K and 55K polypeptides were expressed in induced Ramos/AW and superinfected Raji cells. However, none of these EA-D polypeptides was detectable in BJAB/GC, BJAB, Ramos, and mock-infected Raji cells. To approximate relative concentrations of EA-D in cell extracts, we employed the enzyme-linked immunosorbent assay and immunoblot dot methods by using one of the purified EA-D components to construct a standard curve. Depending upon the cell lines, it was estimated that ca. 1 to 3% (determined by the enzyme

  10. A theoretical timeline for myocardial infarction: immunohistochemical evaluation and western blot quantification for Interleukin-15 and Monocyte chemotactic protein-1 as very early markers

    PubMed Central

    2014-01-01

    Background Experimental and human studies have demonstrated that innate immune mechanisms and consequent inflammatory reaction play a critical role in cardiac response to ischemic injury. Thus, the detection of immuno-inflammatory and cellular phenomena accompanying cardiac alterations during the early inflammatory phase of myocardial infarction (MI) may be an excellent diagnostic tool. Current knowledge of the chronology of the responses of myocardial tissue following the occurrence of ischemic insult, as well as the existence of numerous studies aiming to identify reliable markers in dating MI, induced us to investigate the myocardial specimens of MI fatal cases in order to better define the age of MI. Methods We performed an immunohistochemical study and a Western blot analysis to evaluate detectable morphological changes in myocardial specimens of fatal MI cases and to quantify the effects of cardiac expression of inflammatory mediators (CD15, IL-1β, IL-6, TNF-α, IL-15, IL-8, MCP-1, ICAM-1, CD18, tryptase) and structural and functional cardiac proteins. Results We observed a biphasic course of MCP-1: it was strongly expressed in the very early phase (0-4 hrs), to diminish in the early period (after 6-8 hrs). Again, our choice of IL-15 is explained by the synergism with neutrophilic granulocytes (CD15) and our study shows the potential for striking cytokine synergy in promoting fast, local neutrophil response in damaged tissues. A progressively stronger immunoreaction for the CD15 antibody was visible in the areas where the margination of circulating inflammatory cells was detectable, up to very strong expression in the oldest ones (>12 hours). Further, the induction of CD15, IL-15, MCP-1 expression levels was quantified by Western blot analysis. The results were as follows: IL-15/β-actin 0.80, CD15/β-actin 0.30, and MCP-1/β-actin 0.60, matching perfectly with the results of immunohistochemistry. Control hearts from traumatic death cases did not show any

  11. Detection of immunoglobulin M antibodies to glycoprotein G-2 by western blot (immunoblot) for diagnosis of initial herpes simplex virus type 2 genital infections.

    PubMed Central

    Ho, D W; Field, P R; Irving, W L; Packham, D R; Cunningham, A L

    1993-01-01

    Western blots (immunoblots) for the detection of immunoglobulin M (IgM) antibodies specific for herpes simplex virus type 1 (HSV-1) and HSV-2 in patients' sera were developed. The locations of the type-specific glycoprotein G (gpG-2) of HSV-2 (92- and 140-kDa forms) and glycoprotein C of HSV-1 (gpC-1), which carries mostly type-specific antigenic epitopes, were checked with specific monoclonal antibodies. Western blot assays for IgM antibody to gpC-1 or gpG-2 were performed after depletion of IgG by precipitation with anti-human IgG. In patients with primary HSV-2 genital infections, seroconversion of IgM and IgG antibodies to both the 92- and 140-kDa forms of gpG-2 was observed, although both antibodies appeared in convalescent-phase serum after the first week. IgM and IgG antibodies to low-molecular-size polypeptides (40 to 65 kDa) were the first antibodies observed in patients with primary infection, but these antibodies were cross-reactive with HSV-1 and HSV-2. However, in patients with recurrent HSV-2 infections, IgG antibodies to both forms of gpG-2 and the low-molecular-size polypeptides were found no matter how early after onset the patient was bled, and IgM to gpG-2 did not appear. In patients with nonprimary initial genital HSV-2 infections, IgG antibody to HSV-1 was demonstrated in the first serum specimen, and HSV-2-specific IgM was found in 39% of the serum specimens. Hence, the Western blot assay can be used to test for IgM antibody to gpG-2, allowing for the retrospective diagnosis of inital HSV-2 infections and its use as a supplementary test to the gpG-2 IgG enzyme-linked immunosorbent assays developed elsewhere. In contrast, IgM antibody to gpG-2 is not usually detected in patients with recurrent HSV-2 infections. Images PMID:7508453

  12. 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

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

    PubMed

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

    2015-08-01

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

  14. Multiplexed quantification of plant thylakoid proteins on Western blots using lanthanide-labeled antibodies and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS).

    PubMed

    de Bang, Thomas Christian; Pedas, Pai; Schjoerring, Jan Kofod; Jensen, Poul Erik; Husted, Søren

    2013-05-21

    We have developed a novel calibration method that allows concurrent quantification of multiple proteins by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) after Western blotting. Calibrants were made of nitrocellulose membranes doped with lanthanide standards. Excellent linearity was obtained in the interval from 0 to 24 ng lanthanide cm(-2). Cerium-labeled lysozyme was introduced as an internal reference protein, enabling correction for up to 50% difference in transfer efficiency during the blotting of membranes. The sensitivity of the LA-ICP-MS method was comparable to state-of-the-art chemiluminescence detection and was further improved by a factor of 20, using a polymer tag. Our method allowed reproducible and multiplexed quantification of five thylakoid proteins extracted from chloroplasts of the plant species Arabidopsis thaliana (relative standard deviation (RSD) of ≤ 5% in three independent analytical series). The method was capable of measuring the L subunit in photosystem I of an Arabidopsis mutant containing <5% of this particular protein, relative to the wild type. We conclude that the developed calibration method is highly suited for multiplexed and comparative protein studies, allowing for intermembrane comparisons with high sensitivity and reproducibility.

  15. GAPDH and β-actin protein decreases with aging, making Stain-Free technology a superior loading control in Western blotting of human skeletal muscle.

    PubMed

    Vigelsø, Andreas; Dybboe, Rie; Hansen, Christina Neigaard; Dela, Flemming; Helge, Jørn W; Guadalupe Grau, Amelia

    2015-02-01

    Reference proteins (RP) or the total protein (TP) loaded is used to correct for uneven loading and/or transfer in Western blotting. However, the signal sensitivity and the influence of physiological conditions may question the normalization methods. Therefore, three widely used reference proteins [β-actin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and α-tubulin], as well as TP loaded measured by Stain-Free technology (SF) as normalization tool were tested. This was done using skeletal muscle samples from men subjected to physiological conditions often investigated in applied physiology where the intervention has been suggested to impede normalization (ageing, muscle atrophy, and different muscle fiber type composition). The linearity of signal and the methodological variation coefficient was obtained. Furthermore, the inter- and intraindividual variation in signals obtained from SF and RP was measured in relation to ageing, muscle atrophy, and different muscle fiber type composition, respectively. A stronger linearity of SF and β-actin compared with GAPDH and α-tubulin was observed. The methodological variation was relatively low in all four methods (4-11%). Protein level of β-actin and GAPDH was lower in older men compared with young men. In conclusion, β-actin, GAPDH, and α-tubulin may not be used for normalization in studies that include subjects with a large age difference. In contrast, the RPs may not be affected in studies that include muscle wasting and differences in muscle fiber type. The novel SF technology adds lower variation to the results compared with the existing methods for correcting for loading inaccuracy in Western blotting of human skeletal muscle in applied physiology.

  16. Development and validation of a novel protein extraction methodology for quantitation of protein expression in formalin-fixed paraffin-embedded tissues using western blotting.

    PubMed

    Nirmalan, Niroshini J; Harnden, Patricia; Selby, Peter J; Banks, Rosamonde E

    2009-03-01

    The development of efficient formaldehyde cross-link reversal strategies will make the vast diagnostic tissue archives of pathology departments amenable to prospective and retrospective translational research, particularly in biomarker-driven proteomic investigations. Heat-induced antigen retrieval strategies (HIARs) have achieved varying degrees of cross-link reversal, potentially enabling archival tissue usage for proteomic applications outside its current remit of immunohistochemistry (IHC). While most successes achieved so far have been based on retrieving tryptic peptide fragments using shot-gun proteomic approaches, attempts at extracting full-length, non-degraded, immunoreactive proteins from archival tissue have proved challenging. We have developed a novel heat-induced antigen retrieval strategy using SDS-containing Laemmli buffer for efficient intact protein recovery from formalin-fixed tissues for subsequent analysis by western blotting. Protocol optimization and comparison of extraction efficacies with frozen tissues and current leader methodology is presented. Quantitative validation of methodology was carried out in a cohort of matched tumour/normal, frozen/FFPE renal tissue samples from 10 patients, probed by western blotting for a selected panel of seven proteins known to be differentially expressed in renal cancer. Our data show that the protocol enables efficient extraction of non-degraded, full-length, immunoreactive protein, with tumour versus normal differential expression profiles for a majority of the panel of proteins tested being comparable to matched frozen tissue controls (rank correlation, r = 0.7292, p < 1.825e-09). However, the variability observed in extraction efficacies for some membrane proteins emphasizes the need for cautious interpretation of quantitative data from this subset of proteins. The method provides a viable, cost-effective quantitative option for the validation of potential biomarker panels through a range of clinical

  17. A new and frequent human T-cell leukemia virus indeterminate Western blot pattern: epidemiological determinants and PCR results in central African inhabitants.

    PubMed

    Filippone, Claudia; Bassot, Sylviane; Betsem, Edouard; Tortevoye, Patricia; Guillotte, Micheline; Mercereau-Puijalon, Odile; Plancoulaine, Sabine; Calattini, Sara; Gessain, Antoine

    2012-05-01

    Human T-cell leukemia virus (HTLV) indeterminate Western blot (WB) serological patterns are frequently observed in plasma/serum from persons living in intertropical areas. In the framework of ongoing projects on HTLV-1/2 and related viruses in Central Africa, we systematically analyzed plasma from villagers living in South Cameroon by WB. The group included 1,968 individuals (mean age, 44 years; age range, 5 to 90 years; 978 women/990 men), both Bantus (1,165) and Pygmies (803). Plasma samples were tested by WB analysis (MPD HTLV Blot 2.4) and interpreted according to the manufacturer's instructions. Only clear bands were considered in the analysis. Among the 1,968 plasma samples, 38 (1.93%) were HTLV-1, 13 (0.66%) were HTLV-2, and 6 (0.3%) were HTLV WB seropositive. Furthermore, 1,292 (65.65%) samples were WB sero-indeterminate, including 104 (5.28%) with an HTLV-1 Gag-indeterminate pattern (HGIP) and 68 (3.45%) with a peculiar yet unreported pattern exhibiting mostly a strong shifted GD21 and a p28. The other 619 (31.45%) samples were either WB negative or exhibited other patterns, mostly with unique p19 or p24 bands. DNA, extracted from peripheral blood buffy coat, was subjected to PCR using several primer pairs known to detect HTLV-1/2/3/4. Most DNAs from HTLV-1- and HTLV-seropositive individuals were PCR positive. In contrast, all the others, from persons with HTLV-2, HGIP, new WB, and other indeterminate patterns, were PCR negative. Epidemiological determinant analysis of the persons with this new peculiar WB pattern revealed that seroprevalence was independent from age, sex, or ethnicity, thus resembling the indeterminate profile HGIP rather than HTLV-1. Moreover, this new pattern persists over time.

  18. Western blot seroindeterminate individuals for human T-lymphotropic virus I/II (HTLV-I/II) in Fortaleza (Brazil): a serological and molecular diagnostic and epidemiological approach.

    PubMed

    Santos, Terezinha de Jesus Teixeira; Costa, Carlos Maurício de Castro; Goubau, Patrick; Vandamme, Anne-Mieke; Desmyter, Jan; Van Doren, Sonia; Mota, Rosa M S; de Castro Costa, Francine Bovy; Oliveira, Ana C S; Barreto, Vania; Gomes, A F; Carneiro-Proietti, Anna B; de Bruin, Veralice Meireles Sales; de Sousa, Francisca C F; Oriá, Reinaldo Barreto

    2003-06-01

    How to handle Western blot (WB) seroindeterminate individuals for Human T-lymphotropic Virus 1/2 (HTLV-1/2) constitutes a challenge for blood banks and families. We made a cross-sectional study of 191 enzyme linked immunoassay (EIA) reactive individuals from the hematological center (HEMOCE) of Fortaleza (Brazil), examining their serological (WB) and molecular (PCR) diagnosis, and demographic profiles, as well as a possible association of their condition with other infectious pathologies and risk factors. Ethical institutional approval and personal consent were obtained. Out of 191 EIA reactive individuals, 118 were WB seroindeterminate and 73 were seropositive for HTLV-1/2. In the PCR analysis of 41 WB seroindeterminate individuals, 9 (22%) were positive and 32 (78%) were negative for HTLV-1/2. The demographic analysis indicated a trend towards a predominance of males among the seroindeterminate individuals and females in the seropositive ones. The seroindeterminate individuals were younger than the seropositive ones. We did not find any association of these conditions with syphilis, Chagas disease or HIV or hepatitis, and with risk factors such as breast-feeding, blood transfusion, STD (syphilis) and IDU.

  19. Differential staining of Western blots of human secreted glycoproteins from serum, milk, saliva, and seminal fluid using lectins displaying diverse sugar specificities.

    PubMed

    Gilboa-Garber, Nechama; Lerrer, Batya; Lesman-Movshovich, Efrat; Dgani, Orly

    2005-12-01

    Human milk, serum, saliva, and seminal fluid glycoproteins (gps) nourish and protect newborn and adult tissues. Their saccharides, which resemble cell membrane components, may block pathogen adhesion and infection. In the present study, they were examined by a battery of lectins from plants, animals, and bacteria, using hemagglutination inhibition and Western blot analyses. The lectins included galactophilic ones from Aplysia gonad, Erythrina corallodendron, Maclura pomifera (MPL), peanut, and Pseudomonas aeruginosa (PA-IL); fucose-binding lectins from Pseudomonas aeruginosa (PA-IIL), Ralstonia solanacearum (RSL), and Ulex europaeus (UEA-I), and mannose/glucose-binding Con A. The results demonstrated the chosen lectin efficiency for differential analysis of human secreted gps as compared to CBB staining. They unveiled the diversity of these body fluid gp glycans (those of the milk and seminal fluid being highest): the milk gps interacted most strongly with PA-IIL, followed by RSL; the saliva gps with RSL, followed by PA-IIL and MPL; the serum gps with Con A and MPL, followed by PA-IIL and RSL, and the seminal plasma gps with RSL and MPL, followed by UEA-I and PA-IIL. The potential usage of these lectins as probes for scientific, industrial, and medical purposes, and for quality control of the desired gps is clearly indicated.

  20. Estimation of hepatitis E virus (HEV) pig seroprevalence using ELISA and Western blot and comparison between human and pig HEV sequences in Belgium.

    PubMed

    Thiry, Damien; Mauroy, Axel; Saegerman, Claude; Thomas, Isabelle; Wautier, Magali; Miry, Cora; Czaplicki, Guy; Berkvens, Dirk; Praet, Nicolas; van der Poel, Wim; Cariolet, Roland; Brochier, Bernard; Thiry, Etienne

    2014-08-27

    Zoonotic transmission of hepatitis E virus (HEV) is of special concern, particularly in high income countries were waterborne infections are less frequent than in developing countries. High HEV seroprevalences can be found in European pig populations. The aims of this study were to obtain prevalence data on HEV infection in swine in Belgium and to phylogenetically compare Belgian human HEV sequences with those obtained from swine. An ELISA screening prevalence of 73% (95% CI 68.8-77.5) was determined in Belgian pigs and a part of the results were re-evaluated by Western blot (WB). A receiver operating characteristic curve analysis was performed and scenarios varying the ELISA specificity relative to WB were analysed. The seroprevalences estimated by the different scenarios ranged between 69 and 81% and are in agreement with the high exposure of the European pig population to HEV. Pig HEV sequences were genetically compared to those detected in humans in Belgium and a predominance of genotype 3 subtype f was shown in both swine and humans. The high HEV seroprevalence in swine and the close phylogenetic relationships between pig and human HEV sequences further support the risk for zoonotic transmission of HEV between humans and pigs.

  1. Human T lymphotropic virus types I and II western blot seroindeterminate status and its association with exposure to prototype HTLV-I.

    PubMed

    Yao, Karen; Hisada, Michie; Maloney, Elizabeth; Yamano, Yoshihisa; Hanchard, Barrie; Wilks, Rainford; Rios, Maria; Jacobson, Steven

    2006-02-01

    Human T lymphotropic virus types I and II (HTLV-I/II) Western blot (WB) seroindeterminate status, which is defined as an incomplete banding pattern of HTLV protein Gag (p19 or p24) or Env (GD21 or rgp46), is commonly observed. To investigate the significance of this finding, we examined HTLV-I/II serostatus and HTLV-I proviral load in 2 groups of individuals with WB seroindeterminate status. Low proviral loads were detected in 42% of patients with neurologic symptoms and 44% of voluntary blood donors. These data suggest that a subset of WB seroindeterminate individuals may be infected with prototype HTLV-I. To confirm this hypothesis, we evaluated HTLV-I/II serostatus and proviral load in prospectively collected specimens from 66 WB seronegative patients who had received HTLV-I-infected blood products by transfusion. Eight individuals developed WB seroindeterminate profiles after the transfusion. In addition, using a human leukocyte antigen type A*201-restricted HTLV-I Tax11-19 tetramer, we detected virus-specific CD8(+) T cells in peripheral blood mononuclear cells from WB seroindeterminate patients. These CD8(+) T cells were effective at targeting HTLV-I-infected cells. Collectively, the results suggest that HTLV-I/II WB seroindeterminate status may reflect a history of HTLV-I exposure. Our findings warrant further investigation of the possible clinical outcomes associated with WB seroindeterminate status.

  2. An optimized xylene-free protein extraction method adapted to formalin-fixed paraffin embedded tissue sections for western blot analysis.

    PubMed

    Mansour, Anthony G; Khalil, Pamela Abou; Bejjani, Noha; Chatila, Rajaa; Dagher-Hamalian, Carole; Faour, Wissam H

    2017-03-01

    Deparaffinization of formalin-fixed paraffin embedded (FFPE) tissues with xylene currently remains a major challenge to the biomedical community. We developed an efficient xylene-free protocol to isolate proteins from archived FFPE human tissue sections. A total of 79 different types of FFPE tissue sections of 8 µm thickness were obtained from various archived FFPE specimens. Deparaffinization was conducted by gently washing each section with around 1 ml of hot distilled water (≈80°C). The deparaffinized tissues were homogenized in lysis buffer, and the isolated proteins were quantified and efficiently resolved using western blot analysis for the presence of Protein kinase B (PKB/AKT) and β-actin. Moreover, a significant amount of proteins was successfully isolated with an average of 2.31 µg/µl. The migration pattern of AKT and β-actin obtained from the specimens was similar to the positive control obtained from protein lysates prepared from in vitro cultured MDA231 cancer cell lines. AKT was successfully identified in all specimens, and β-actin protein was resolved with an efficiency higher than 80%. The entire extraction procedure requires only 20 minutes. This newly developed technique is an efficient, safe, cost-effective, and rapid method to isolate proteins from FFPE tissue sections adequate for molecular analysis.

  3. Immunohistochemical and Western blot analysis of two protein tyrosine phosphatase receptors, R and Z1, in colorectal carcinoma, colon adenoma and normal colon tissues.

    PubMed

    Woźniak, Marta; Gamian, Elżbieta; Łaczmańska, Izabela; Sąsiadek, Maria M; Duś-Szachniewicz, Kamila; Ziółkowski, Piotr

    2014-05-01

    Two classes of proteins, namely tyrosine kinases (PTK) and phosphatases (PTP), play an important role in cell proliferation and differentiation, thus leading to an acceleration or inhibition of tumour growth. The role of the above proteins in colorectal carcinoma (CRC) growth is a well-known event. In this study we carried out immunohistochemical and Western blot analysis of colorectal carcinoma, adenoma and normal colon tissue in relation to two protein tyrosine phosphatase receptors, R and Z1. Twenty-five cases of CRC were analyzed and the results were compared with similar data obtained in non-malignant tissues. High expression of both PTP receptors was observed in all examined cases of CRC, adenoma and normal colon tissue in this study. These results are not in line with recently published data, showing that genetic coding for PTPRR and PTPRZ1 were hypermethylated in CRC's. We presume that the protein tyrosine phosphatase overexpression in colorectal carcinoma is not enough to protect from the progression of disease.

  4. Anti-LKM-1 antibodies determined by use of recombinant P450 2D6 in ELISA and western blot and their association with anti-HCV and HCV-RNA.

    PubMed Central

    Seelig, R; Renz, M; Bünger, G; Schröter, H; Seelig, H P

    1993-01-01

    Several subtypes of anti-liver-kidney microsome antibodies (LKM) are known. LKM-1 antibodies associated with autoimmune chronic active hepatitis recognize P450 2D6, a cytochrome P450 mono-oxygenase. The frequent association of anti-LKM-1 antibodies and hepatitis C virus (HCV) infections and the probable existence of an infectious and autoimmune form of anti-LKM-1-associated hepatitis, requiring different therapeutical strategies, necessitates the exact determination of anti-LKM-1 specificities. Therefore, we compared various antibody tests (immunofluorescence, ELISA with recombinant P450 2D6, and Western blot with recombinant and natural antigens and agargel double diffusion) with sera of 27 anti-LKM-1-positive chronic active hepatitis (CAH) patients, with 61 sera harbouring anti-mitochondrial antibodies, 100 sera each from HCV-RNA-positive and HCV-RNA-negative patients, and 50 sera of healthy persons. Western blot techniques using recombinant MS2-polymerase P450 2D6 fusion protein were found to be the most sensitive and specific method for anti-LKM-1 antibody determination in routine laboratory. The recently recognized association of anti-LKM-1 antibody and HCV infection was confirmed by the results of this study. In anti-HCV and HCV-RNA-positive patients with anti-LKM-1 antibodies there was a preponderance of males with higher mean age and lower antibody titres. The results support the hypothesis of the existence of an autoimmune as well as an infectious (HCV triggered) subgroup of anti-LKM-1-positive hepatitis. Images Fig. 1 PMID:8390333

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

    PubMed

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

    2015-01-01

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

  6. Human T-Cell Lymphotropic Virus Type 1 Gag Indeterminate Western Blot Patterns in Central Africa: Relationship to Plasmodium falciparum Infection

    PubMed Central

    Mahieux, Renaud; Horal, Peter; Mauclère, Philippe; Mercereau-Puijalon, Odile; Guillotte, Micheline; Meertens, Laurent; Murphy, Edward; Gessain, Antoine

    2000-01-01

    To gain insight on the significance of human T-cell lymphotropic virus type 1 (HTLV-1) indeterminate serological reactivities, we studied villagers of South Cameroon, focusing on a frequent and specific HTLV-1 Gag indeterminate profile (HGIP) pattern (gag p19, p26, p28, and p30 without p24 or Env gp21 and gp46). Among the 102 sera studied, 29 from all age groups had a stable HGIP pattern over a period of 4 years. There was no epidemiological evidence for sexual or vertical transmission of HGIP. Seventy-five percent of HGIP sera reacted positively on MT2 HTLV-1-infected cells by immunofluorescence assay. However, we could not isolate any HTLV-1 virus or detect the presence of p19 Gag protein in cultures of peripheral blood mononuclear cells obtained from individuals with strong HGIP reactivity. PCR experiments conducted with primers for HTLV-1 and HTLV-2 (HTLV-1/2 primers) encompassing different regions of the virus did not yield HTLV-1/2 proviral sequences from individuals with HGIP. Using 11 peptides corresponding to HTLV-1 or HTLV-2 immunodominant B epitopes in an enzyme-linked immunosorbent assay, one epitope corresponding to the Gag p19 carboxyl terminus was identified in 75% of HGIP sera, while it was recognized by only 41% of confirmed HTLV-1-positive sera. A positive correlation between HTLV-1 optical density values and titers of antibody to Plasmodium falciparum was also demonstrated. Finally, passage of sera through a P. falciparum-infected erythrocyte-coupled column was shown to specifically abrogate HGIP reactivity but not the HTLV-1 pattern, suggesting the existence of cross-reactivity between HTLV-1 Gag proteins and malaria-derived antigens. These data suggest that in Central Africa, this frequent and specific Western blot is not caused by HTLV-1 infection but could instead be associated with P. falciparum infection. PMID:11060067

  7. 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

  8. Identification in gelada baboons (Theropithecus gelada) of a distinct simian T-cell lymphotropic virus type 3 with a broad range of Western blot reactivity.

    PubMed

    Van Dooren, Sonia; Shanmugam, Vedapuri; Bhullar, Vinod; Parekh, Bharat; Vandamme, Anne-Mieke; Heneine, Walid; Switzer, William M

    2004-02-01

    Antibodies to simian T-cell lymphotropic virus (STLV) were found in serum or plasma from 12 of 23 (52.2 %) gelada baboons (Theropithecus gelada) captive in US zoos. A variety of Western blot (WB) profiles was seen in the 12 seroreactive samples, including human T-cell lymphotropic virus (HTLV)-1-like (n=5, 41.7 %), HTLV-2-like (n=1, 8.3 %), HTLV-untypable (n=4, 33.3 %) and indeterminate (n=2, 16.6 %) profiles. Phylogenetic analysis of tax or env sequences that had been PCR amplified from peripheral blood lymphocyte DNA available from nine seropositive geladas showed that four were infected with identical STLV-1s; these sequences clustered with STLV-1 from Celebes macaques and probably represent recent cross-species infections. The tax sequences from the five remaining geladas were also identical and clustered with STLV-3. Analysis of the complete STLV-3 genome (8917 bp) from one gelada, TGE-2117, revealed that it is unique, sharing only 62 % similarity with HTLV-1/ATK and HTLV-2/Mo. STLV-3/TGE-2117 was closest genetically to STLV-3 from an Eritrean baboon (STLV-3/PH969, 95.6 %) but more distant from STLV-3s from red-capped mangabeys from Cameroon and Nigeria (STLV-3/CTO-604, 87.7 %, and STLV-3/CTO-NG409, 87.2 %, respectively) and Senegalese baboons (STLV-3/PPA-F3, 88.4 %). The genetic relatedness of STLV-3/TGE-2117 to STLV-3 was confirmed by phylogenetic analysis of a concatenated gag-pol-env-tax sequence (6795 bp). An ancient origin of 73 628-109 809 years ago for STLV-3 was estimated by molecular clock analysis of third-codon positions of gag-pol-env-tax sequences. LTR sequences from five STLV-3-positive geladas were >99 % identical and clustered with that from a Papio anubisxP. hamadryas hybrid Ethiopian baboon, suggesting a common source of STLV-3 in these sympatric animals. LTR sequences obtained 20 years apart from a mother-infant pair were identical, providing evidence of both mother-to-offspring transmission and a high genetic stability of STLV-3. Since

  9. Comparison of the efficiency of two commercial kits - ELFA and Western blot in estimating the phase of Toxoplasma gondii infection in pregnant women.

    PubMed

    Sroka, Jacek; Wójcik-Fatla, Angelina; Zając, Violetta; Sawczyn, Anna; Cisak, Ewa; Karamon, Jacek; Dutkiewicz, Jacek; Bojar, Iwona

    2016-12-23

    Sera of 89 pregnant women were selected according to the results of ELFA IgM, IgG and avidity IgG, and tested with commercial tests IgM, IgG and avidity IgG Western Blot (WB) to compare the efficacy of both techniques in determining the phase of T. gondii infection. In total, 81 of 89 tested sera (91.0%) were classified as positive, both in the ELFA and WB tests for the presence of anti-Toxoplasma antibodies of class IgG, indicating a past infection, while the prevalence of anti-Toxoplasma positive reactions associated with the antibodies of class IgM indicating a recent infection was much lower - 31.5% and 20.2%, respectively. Sera of 81 women were also tested in the ELFA and WB tests for avidity, e.g. ability of forming high-molecular IgG antibody complexes. Low or medium results in these tests (in this study all classified as low), indicating a recent infection, were detected by ELFA and WB in 22.2% and 45.7% of the total examined samples, respectively. The Spearman's rank test for correlation, performed for recognition of quantitative data of the ELFA and WB tests (index, units or points), revealed a highly significant correlation between the ELFA and WB tests for homologous classes of antibodies, both for IgM and IgG (p<0.00001). In contrast, the ELFA and WB tests for detection of anti-Toxoplasma IgM antibodies were not correlated with the ELFA and WB tests for detection of anti-Toxoplasma IgG antibodies (p>0.05), except for the WB test for IgM antibodies, which showed a significant correlation with the ELFA test for IgG antibodies (p<0.01). A highly significant negative correlation between the ELFA and WB test for IgM antibodies and ELFA and WB tests for IgG avidity was demonstrated (p<0.01), except for a relationship between the WB test for IgM and WB for avidity, which was not significant. Such negative correlations are theoretically expected, as strong complexes with the participation of IgG antibodies are absent in the early phase of toxoplasmosis when

  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.

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

    PubMed

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

    1995-04-01

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

  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. Identification of liver protein targets modified by tienilic acid metabolites using a two-dimensional Western blot-mass spectrometry approach

    NASA Astrophysics Data System (ADS)

    Methogo, Ruth Menque; Dansette, Patrick M.; Klarskov, Klaus

    2007-12-01

    A combined approach based on two-dimensional electrophoresis-immuno-blotting and nanoliquid chromatography coupled on-line with electrospray ionization mass spectrometry (nLC-MS/MS) was used to identify proteins modified by a reactive intermediate of tienilic acid (TA). Liver homogenates from rats exposed to TA were fractionated using ultra centrifugation; four fractions were obtained and subjected to 2D electrophoresis. Following transfer to PVDF membranes, modified proteins were visualized after India ink staining, using an anti-serum raised against TA and ECL detection. Immuno-reactive spots were localized on the PVDF membrane by superposition of the ECL image, protein spots of interest were excised, digested on the membrane with trypsin followed by nLC-MS/MS analysis and protein identification. A total of 15 proteins were identified as likely targets modified by a TA reactive metabolite. These include selenium binding protein 2, senescence marker protein SMP-30, adenosine kinase, Acy1 protein, adenosylhomocysteinase, capping protein (actin filament), protein disulfide isomerase, fumarylacetoacetase, arginase chain A, ketohexokinase, proteasome endopeptidase complex, triosephosphate isomerase, superoxide dismutase, dna-type molecular chaperone hsc73 and malate dehydrogenase.

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

    SciTech Connect

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

    1987-05-01

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

  15. Negative correlation between poly-ADP-ribosylation of spleen cell histone proteins and initial duration of dimethylnitrosamine exposure to mice in vivo measured by Western blot immunoprobe assay: a possible biomarker for cancer detection.

    PubMed

    Devi, Brahmacharimayum J; Schneeweiss, Frank H A; Sharan, Rajeshwar N

    2005-01-01

    Improved cancer detection involving suitable biomarkers with easy applicability is a challenge to our fight against cancer. Poly-ADP-ribosylation (PAR) of proteins is a likely candidate biomarker for this purpose because it meets the criterion well. This report is a step towards testing suitability of PAR as a biomarker for cancer detection. Swiss albino mice were exposed to hepatocarcinogen, dimethylnitrosamine (DMN), at a chronic dose, which is known to induce carcinogenesis in liver. PAR was monitored by a Western blot immunoprobe assay in spleen, a lymphoid organ, to find a correlation between PAR of spleen histone proteins and duration of DMN exposure. A negative, non-linear correlation was found for most histone proteins. The inhibition of PAR of histones was significant from 4 weeks onwards until the end of the observation. The inhibition was potentiated when 3-aminobenzamide was simultaneously administered. The results open up the possibility of PAR of cellular proteins being used as biomarker for cancer detection.

  16. Detection and quantification of poly-ADP-ribosylated cellular proteins of spleen and liver tissues of mice in vivo by slot and Western blot immunoprobing using polyclonal antibody against mouse ADP-ribose polymer.

    PubMed

    Sharan, R N; Devi, B Jaylata; Humtsoe, J O; Saikia, Jyoti R; Kma, L

    2005-10-01

    Poly-ADP-ribosylation (PAR) of cellular proteins has been shown to have decisive roles in diverse cellular functions including carcinogenesis. There are indications that metabolic level of poly-ADP-ribosylated cellular proteins might indicate carcinogenesis and, therefore, could be potentially used in cancer screening program. Keeping in mind the limitations of currently available assays of cellular PAR, a new assay is being reported that measures the metabolic level of poly-ADP-ribosylated cellular proteins. The ELISA based slot and Western blot immunoassay used polyclonal antibody against natural, heterogeneous ADP-ribose polymers. It could be successfully employed to qualitatively and quantitatively assay metabolic levels of poly-ADP-ribosylated proteins of spleen and liver tissues of normal mice or mice exposed to dimethylnitrosamine for up to 8 weeks; potentially PAR of cellular proteins could be assayed in any tissue or biopsy. Implications of the results in cancer screening program have been discussed.

  17. ELISA/Western blot tests for HIV

    MedlinePlus

    ... R, Blaser MJ, eds. Mandell, Douglas, and Bennett's Principles and Practice of Infectious Diseases . 8th ed. Philadelphia, PA: Elsevier Saunders; 2015:chap 122. U.S. Preventive Services Task Force. Final Update Summary: Human Immunodeficiency Virus ( ...

  18. 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.

  19. Neither molecular diversity of the envelope, immunosuppression status, nor proviral load causes indeterminate HTLV western blot profiles in samples from human T-cell lymphotropic virus type 2 (HTLV-2)-infected individuals.

    PubMed

    Olah, Ingrid; Fukumori, Ligia M I; Smid, Jerusa; de Oliveira, Augusto César Penalva; Duarte, Alberto J S; Casseb, Jorge

    2010-05-01

    Although human T-cell lymphotropic virus type 2 (HTLV-2) is considered of low pathogenicity, serological diagnosis is important for counseling and monitoring. The confirmatory tests most used are Western blot (WB) and PCR. However, in high-risk populations, about 50% of the indeterminate WB were HTLV-2 positives by PCR. The insensitivity of the WB might be due to the use of recombinant proteins of strains that do not circulate in our country. Another possibility may be a high level of immunosuppression, which could lead to low production of virus, resulting in low stimulation of antibody. We found one mutation, proline to serine in the envelope region in the position 184, presented at least 1/3 of the samples, independent the indeterminate WB profile. In conclusion, we found no correlation of immune state, HTLV-2 proviral load, or env diversity in the K55 region and WB indeterminate results. We believe that the only WB kit available in the market is probably more accurate to detect HTLV-1 antibodies, and some improvement for HTLV-2 detection should be done in the future, especially among high-risk population.

  20. Distribution of homologous proteins to puffer fish saxitoxin and tetrodotoxin binding protein in the plasma of puffer fish and among the tissues of Fugu pardalis examined by Western blot analysis.

    PubMed

    Yotsu-Yamashita, Mari; Yamaki, Hiroe; Okoshi, Natsumi; Araki, Nao

    2010-06-01

    Puffer fish saxitoxin and tetrodotoxin binding protein (PSTBP) is a glycoprotein (200 kDa as a dimer) that we previously isolated from the plasma of Fugu pardalis (Yotsu-Yamashita et al., 2001). For the study on functions of PSTBP, here we examined distribution of homologous proteins to PSTBP in the plasma of seven species of puffer fish, and among the tissues of F. pardalis by Western blot analysis probed with a polyclonal IgG against unglycosylated PSTBP1 expressed in Echelichia coli. One or two major positive broad bands were detected at 105-140 kDa molecular weight range in the plasma (0.5 microg protein) of all species of puffer fish tested, while no band was detected in the plasma (5 microg protein) of fish other than puffer fish. Glycopeptidase F treated plasma of all species of puffer fish tested commonly showed the bands at approximately 42 kDa that was consistent to the molecular weight of unglycosylated PSTBP. These data suggest that puffer fish commonly possess glycoproteins homologous to PSTBP, but the sizes of N-glycan are specific to the species. Among soluble protein extracts (5 microg protein) from the tissues of F. pardalis, PSTBP was detected in all tissues examined, most prominently in heart, skin, and gall.

  1. Serological, epidemiological, and molecular differences between human T-cell lymphotropic virus Type 1 (HTLV-1)-seropositive healthy carriers and persons with HTLV-I Gag indeterminate Western blot patterns from the Caribbean.

    PubMed

    Rouet, F; Meertens, L; Courouble, G; Herrmann-Storck, C; Pabingui, R; Chancerel, B; Abid, A; Strobel, M; Mauclere, P; Gessain, A

    2001-04-01

    To investigate the significance of serological human T-cell lymphotropic virus type 1 (HLTV-1) Gag indeterminate Western blot (WB) patterns in the Caribbean, a 6-year (1993 to 1998) cross-sectional study was conducted with 37,724 blood donors from Guadeloupe (French West Indies), whose sera were routinely screened by enzyme immunoassay (EIA) for the presence of HTLV-1 and -2 antibodies. By using stringent WB criteria, 77 donors (0.20%) were confirmed HTLV-1 seropositive, whereas 150 (0.40%; P < 0.001) were considered HTLV seroindeterminate. Among them, 41.3% (62) exhibited a typical HTLV-1 Gag indeterminate profile (HGIP). Furthermore 76 (50.7%) out of the 150 HTLV-seroindeterminate subjects were sequentially retested, with a mean duration of follow-up of 18.3 months (range, 1 to 70 months). Of these, 55 (72.4%) were still EIA positive and maintained the same WB profile whereas the others became EIA negative. This follow-up survey included 33 persons with an HGIP. Twenty-three of them (69.7%) had profiles that did not evolve over time. Moreover, no case of HTLV-1 seroconversion could be documented over time by studying such sequential samples. HTLV-1 seroprevalence was characterized by an age-dependent curve, a uniform excess in females, a significant relation with hepatitis B core (HBc) antibodies, and a microcluster distribution along the Atlantic coast of Guadeloupe. In contrast, the persons with an HGIP were significantly younger, had a 1:1 sex ratio, did not present any association with HBc antibodies, and were not clustered along the Atlantic façade. These divergent epidemiological features, together with discordant serological screening test results for subjects with HGIP and with the lack of HTLV-1 proviral sequences detected by PCR in their peripheral blood mononuclear cell DNA, strongly suggest that an HGIP does not reflect true HTLV-1 infection. In regard to these data, healthy blood donors with HGIP should be reassured that they are unlikely to be

  2. 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

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

    NASA Astrophysics Data System (ADS)

    Choi, Ki-Seon; Moon, Il-Ju

    2012-09-01

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

  5. Detection of Proteins on Blot Membranes.

    PubMed

    Goldman, Aaron; Harper, Sandra; Speicher, David W

    2016-11-01

    Staining of blot membranes enables the visualization of bound proteins. Proteins are usually transferred to blot membranes by electroblotting, by direct spotting of protein solutions, or by contact blots. Staining allows the efficiency of transfer to the membrane to be monitored. This unit describes protocols for staining proteins after electroblotting from polyacrylamide gels to blot membranes such as polyvinylidene difluoride (PVDF), nitrocellulose, or nylon membranes. The same methods can be used if proteins are directly spotted, either manually or using robotics. Protocols are included for seven general protein stains (amido black, Coomassie blue, Ponceau S, colloidal gold, colloidal silver, India ink, and MemCode) and three fluorescent protein stains (fluorescamine, IAEDANS, and SYPRO Ruby). Also included is an in-depth discussion of the different blot membrane types and the compatibility of different protein stains with downstream applications, such as immunoblotting or N-terminal Edman sequencing. © 2016 by John Wiley & Sons, Inc.

  6. Fault activation by hydraulic fracturing in western Canada.

    PubMed

    Bao, Xuewei; Eaton, David W

    2016-12-16

    Hydraulic fracturing has been inferred to trigger the majority of injection-induced earthquakes in western Canada, in contrast to the Midwestern United States, where massive saltwater disposal is the dominant triggering mechanism. A template-based earthquake catalog from a seismically active Canadian shale play, combined with comprehensive injection data during a 4-month interval, shows that earthquakes are tightly clustered in space and time near hydraulic fracturing sites. The largest event [moment magnitude (MW) 3.9] occurred several weeks after injection along a fault that appears to extend from the injection zone into crystalline basement. Patterns of seismicity indicate that stress changes during operations can activate fault slip to an offset distance of >1 km, whereas pressurization by hydraulic fracturing into a fault yields episodic seismicity that can persist for months.

  7. Fault activation by hydraulic fracturing in western Canada

    NASA Astrophysics Data System (ADS)

    Bao, Xuewei; Eaton, David W.

    2016-12-01

    Hydraulic fracturing has been inferred to trigger the majority of injection-induced earthquakes in western Canada, in contrast to the Midwestern United States, where massive saltwater disposal is the dominant triggering mechanism. A template-based earthquake catalog from a seismically active Canadian shale play, combined with comprehensive injection data during a 4-month interval, shows that earthquakes are tightly clustered in space and time near hydraulic fracturing sites. The largest event [moment magnitude (MW) 3.9] occurred several weeks after injection along a fault that appears to extend from the injection zone into crystalline basement. Patterns of seismicity indicate that stress changes during operations can activate fault slip to an offset distance of >1 km, whereas pressurization by hydraulic fracturing into a fault yields episodic seismicity that can persist for months.

  8. One Health training and research activities in Western Europe

    PubMed Central

    Sikkema, Reina; Koopmans, Marion

    2016-01-01

    Introduction The increase in emerging human infectious diseases that have a zoonotic origin and the increasing resistance of microorganisms to antimicrobial drugs have shown the need for collaborations between the human, animal and environmental health sectors. The One Health concept increasingly receives recognition from policy makers and researchers all over the world. This overview compiled research and education activities in the area of One Health in Western Europe (Austria, Belgium, France, Germany, Italy, Iceland, Ireland, Liechtenstein, Luxembourg, Monaco, the Netherlands, Portugal, Scandinavia, Spain, Switzerland, and the United Kingdom (UK), with a focus on infectious diseases. It can serve as a starting point for future initiatives and collaborations. Material and methods A literature search for ‘One Health’ was performed using National Center for Biotechnology Information and Google. Moreover, information from global and European policy documents was collected and a questionnaire was designed to gather current One Health research and training activities in Western Europe. Results This overview shows that there is considerable recognition for One Health in Europe, although most educational initiatives are recent. In Europe, the One Health approach is currently mainly advocated in relation to antimicrobial resistance (AMR). Many countries have incorporated the One Health approach in their policy to fight AMR, and funding possibilities for AMR research increased significantly. The number of national and international multidisciplinary research networks in the area of zoonotic diseases and One Health is increasing. Discussion Although One Health has gained recognition in Europe, often a One Health approach to research and education in the area of zoonotic diseases and AMR is not implemented. In many countries, collaboration between sectors is still lacking, and One Health activities are predominantly initiated by the veterinary sector. To facilitate the

  9. 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

  10. 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

  11. Active tectonics of central-western Caucasus, Georgia

    NASA Astrophysics Data System (ADS)

    Tsereteli, N.; Tibaldi, A.; Alania, V.; Gventsadse, A.; Enukidze, O.; Varazanashvili, O.; Müller, B. I. R.

    2016-11-01

    This work contributes to a better knowledge of potentially seismogenic faults of the Georgia Greater and Lesser Caucasus by evaluating the distribution of earthquake foci, active tectonic stress field, kinematics and geometry of main fault planes. We consider all the information coming from field structural geology, geomorphology, seismological data from historical and instrumental catalogues, seismic reflection sections, as well as new focal mechanism solutions. These data enable recognizing some active ENE-WSW reverse faults in the core of the Greater Caucasus that are parallel to the mountain range. At the southernmost front of the Greater Caucasus, a series of main thrusts dipping towards NNE are active, with up to hundreds-km-long segments; along this thrust zone, a potentially locked segment is present, about 90 km long. The studied section of the Lesser Caucasus has active structures along the northern front given by south-dipping thrusts, as well as in the central core where strike-slip and oblique faults coexist. The Transcaucasian depression between the two mountain ranges shows an ongoing inversion tectonics of the central part of the Rioni Basin where active N- to NE-dipping reverse faults are present, accompanied by clear evidence of uplift of a wide area. The data are coherent with a N-S to NNE-SSW contraction of the central-western Greater Caucasus and Lesser Caucasus. Although in general the seismicity decreases westward in terms of number of earthquakes and magnitude, seismological and geological structural data in the Rioni Basin indicate here a Quaternary propagation of deformation towards the west.

  12. Immunoconfirmation of central nervous system tuberculosis by blotting: A study of 300 cases.

    PubMed

    Patil, Shripad; Giribhattanavar, Prashant; Patil, Madhusudan; Kumar, Kavitha

    2015-06-01

    Tuberculous meningitis (TBM) is a serious form of disease of the central nervous system. Early and accurate diagnosis of the disease and effective treatment are key important factors to contain the disease. The disease presents as chronic meningitis where other partners such as fungal meningitis, neurosyphilis, cysticercal meningitis, carcinomatous meningitis and partially treated pyogenic meningitis share a similar clinical picture making the diagnosis complicated. Culturing of the pathogen Mycobacterium tuberculosis (MTB) from the cerebrospinal fluid (CSF) sample has shown a poor response. The main immunological method for the immunodiagnosis of TBM is the detection of an antibody response in the CSF. In the present study, total MTB sonicated extract antigen was used for ELISA and Western blot. ELISA shows overall immune response of the test sample, whereas Western blotting reveals the specific reactivity to a particular molecular weight antigen. This would also reveal the immunodominant antigen. A total of 300 CSF samples were analyzed by both ELISA and Western blotting. Of the 240 clinically suspected TBM cases, 111 samples were positive by ELISA and 81 samples by Western blot. A total of 76 CSF samples were positive by both ELISA and Western blot. None of the control samples showed positivity either by ELISA or by Western blot. TBM patients revealed major antibody reactivity to 30-40 kD region, followed by 14 kD region. ELISA is sensitive with mild non-specific binding, but Western blot is specific in detecting the immune response. The findings will be useful in definitive immunodiagnosis of TBM.

  13. 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.

  14. Blot hybridisation analysis of genomic DNA.

    PubMed Central

    Vandenplas, S; Wiid, I; Grobler-Rabie, A; Brebner, K; Ricketts, M; Wållis, G; Bester, A; Boyd, C; Måthew, C

    1984-01-01

    Restriction endonuclease analysis of specific gene sequences is proving to be a valuable technique for characterisation and diagnosis of inherited disorders. This paper describes detailed protocols for isolation, restriction, and blot hybridisation of genomic DNA. Problems and alternatives in the procedure are discussed and a troubleshooting guide has been provided to help rectify faults. Images PMID:6086927

  15. TLC-Blot (Far-Eastern Blot) and Its Application to Functional Lipidomics.

    PubMed

    Taki, Takao

    2015-01-01

    A simple method for transfer of lipids-including phospholipids, glycolipids, and neutral lipids-from a high performance thin layer chromatography (HPTLC) plate to a polyvinylidene difluoride (PVDF) membrane, TLC-Blot (Far-Eastern Blot), and its biochemical applications are presented. This chapter presents the conventional procedures for separating lipid from tissue samples, cultured cells, and serum and the subsequent development of TLC. Individual lipids separated on an HPTLC plate can be transferred to the PVDF membrane quantitatively and also isolated from the lipid-blotted membrane by a one-step purification procedure. Immunodetection with monoclonal antibodies and treatment with lipid-metabolizing enzymes on the lipid-blotted membrane are possible. The method for identification of individual lipids transferred on the PVDF membrane using matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometry (TLC-Blot/MALDI-TOF MS) is shown as a functional lipidomics application.

  16. Kinematic indicators on active normal faults in Western Turkey

    NASA Astrophysics Data System (ADS)

    Hancock, P. L.; Barka, A. A.

    Quaternary normal fault zones in western Turkey comprise multiple slip planes and zone-parallel layers of fault breccia. They also contain several little-known kinematic indicators that are probably typical of many formed at shallow levels in extending terrains. The recent exhumation by contractors of about 2000 m 2 of slip planes in a SSE-dipping fault zone separating Quaternary colluvium from bedrock carbonates at Yavansu (7 km SE of Kuşadasi) permits an unusually complete inventory of the indicators to be compiled. The most spectacular indicators are metre-scale 69°W-pitching corrugations in slip planes and recemented breccia sheets underlying them. Corrugations, characterized by sinusoidal profiles normal to their long axes and, less commonly, culminations and depression along their axes possibly developed as a result of upwards-propagating slip planes seeking undemanding pathways through heterogeneous fault-precursor breccias that formed in advance of tip lines. Parallel to corrugation long axes are those of gutters, flat-floored, sleep-sided channels a few centimetres wide, probably related to the abrasion of subslip-plane breccia sheets. Centimetre-scale tool tracks scored in the uppermost subslip-plane breccia sheet by resistant colluvial clasts are irregular at their proximal ends but distally they swing into alignment with corrugation axes. Frictional-wear striae, centimetres long but only a few millimetres wide and pitching 78°W, are superimposed on the other slip-parallel lineations. Comb fractures nearly perpendicular to slip planes define an intersection lineation which is normal to corrugation axes. Fault-plane solutions of earthquakes on SSE-dipping active faults in the West Anatolian extensional province indicate that mainly normal, combined with minor dextral slip is the dominant mode, a conclusion in accord with the sense of slip inferred from the indicators exposed on the Yavansu slip planes.

  17. 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.

  18. Gel electrophoresis of polyphenol oxidase with instant identification by in situ blotting.

    PubMed

    Cheng, Tsai-Mu; Huang, Pei-Chen; Pan, Ju-Pin; Lin, Kuan-Yu; Mao, Simon J T

    2007-04-15

    Polyphenol oxidase (PPO) or tyrosinase is an important and ubiquitous enzyme responsible for browning in plants and melanization in animals. The molecular size of the plant PPO is varied among the species and its activity can be enhanced by a variety of anionic detergents. In the present study, we developed a simple method for the first-step identification of PPO in fruit and vegetable extracts. First, 3mm chromatographic paper was immersed in 0.5% (w/v) catechol solution as an immobilized PPO substrate. After running the extract with 10% sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), one side of the glass plate was removed. The plate was immediately laid on top of the dried catechol-paper. A dark-brown band corresponding to PPO was visualized within 1 min and was further confirmed by a conventional Western blot using an antibody prepared against mushroom PPO. It also reveals that some vegetation (such as tomato, radish, and oriental melon) with low or no detectable activity in a conventional enzyme assay actually possessed marked levels of PPO activity when assessed by PAGE-blot. We propose that an inhibitor is associated with PPO in some plants; the inhibitor, however, is dissociated during the electrophoresis. Therefore, in addition to identify the molecular form of PPO, the present technique may explore the existence of PPO inhibitor(s) in plants. The detail of the method with respect to its relevance for searching a natural PPO inhibitor is described and discussed.

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

    PubMed

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

    2009-02-01

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

  20. 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.

  1. 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…

  2. Repeated probing of Southwestern blots using alkaline phosphatase stripping.

    PubMed

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

    2010-11-05

    Southwestern blotting is when a DNA sequence is used to probe DNA-binding proteins on an electrophoretic gel blot. It would be highly desirable to be able to probe a blot repeatedly with different DNA sequences. Alkaline phosphatase can remove 5'-phosphoryl groups from DNA and radiolabeled 5'-(32)P-DNA probes are commonly used in Southwestern blotting. Here is shown that once probed, the radioisotope signal on the blot can be effectively removed by brief digestion with alkaline phosphatase, and the blot can then be repeatedly probed at least six times with different DNA probes. This exceeds the repetitions possible with another commonly used method using SDS. The technique can be used with either one-dimensional or multi-dimensional Southwestern blots and does not have a large effect on the phosphorylation state of the blotted proteins. An alternative method using T4 polynucleotide kinase stripping is also introduced but was less well characterized.

  3. Cell adhesion to proteins separated by lithium dodecyl sulfate-polyacrylamide gel electrophoresis and blotted onto a polyvinylidene difluoride membrane: a new cell-blotting technique.

    PubMed

    Seshi, B

    1994-12-02

    Cell blotting, although conceptually simple, has failed to achieve wide practical application. Described here is a new cell-blotting technique which involves cell adhesion to protein bands after separation by lithium dodecyl sulfate-polyacrylamide gel electrophoresis (LDS-PAGE) and blotting onto polyvinylidene difluoride (PVDF) membrane at 4 degrees C. Cell bands adherent on PVDF are detected using hematoxylin, or propidium iodide (PI) staining followed by viewing under ultraviolet (UV) light. The technique allows quick microscopic visualization of adherent cells composing the bands, without requiring clearing of the membrane. Representative cell adhesion proteins from different sources, i.e., plant lectins (e.g., phytohemagglutinin, PHA; concanavalin A, ConA; and wheat germ agglutinin, WGA); extracellular matrix (ECM) proteins; and integral membrane proteins (e.g., recombinant soluble vascular cell adhesion molecule-1, rs VCAM-1) were tested for cell binding by the new cell-blotting technique using human lymphoid progenitor (NALM-6) and myeloid progenitor (KG1a) cell lines. Cell adhesion proteins retained their adhesion function in all cases tested. Specificity of cell binding on PVDF blot was demonstrated by inhibition of cell adhesion to WGA protein bands using an appropriate sugar, i.e., N-acetyl D-glucosamine. The cell blotting assay was comparable in sensitivity to Coomassie blue staining of protein bands. The ability to conduct protein extraction, separation and blotting at low temperature avoids thermal denaturation, thereby preserving the adhesion properties of the proteins. The electrophoretic/blotting system has unique detergent removal/protein renaturation properties and the ability to preserve functionally active adhesion protein complexes. The cell-blotting technique described is sufficiently robust for routine application in the investigation of novel cell adhesion proteins.

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

    USGS Publications Warehouse

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

    2006-01-01

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

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

    PubMed Central

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

    2016-01-01

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

  6. 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.

  7. Relationships Between Global Warming and Tropical Cyclone Activity in the Western North Pacific

    DTIC Science & Technology

    2007-09-01

    In this work, we investigate the relationships between global warming and tropical cyclone activity in the Western North Pacific (WNP). Our...hypothesis is that global warming impacts on TC activity occur through changes in the large scale environmental factors (LSEFs) known to be important in...averages. Using a least squares fit, we identify global warming signals in both the SST and vertical wind shear data across the WNP. These signals vary

  8. 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.

  9. 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.

  10. Truncated seasonal activity patterns of the western blacklegged tick (Ixodes pacificus) in central and southern California.

    PubMed

    MacDonald, Andrew J; Briggs, Cheryl J

    2016-02-01

    Patterns of seasonal activity and density of host-seeking western blacklegged ticks, Ixodes pacificus, were investigated in central and southern California. Weekly to monthly drag sampling was undertaken at two sites in Santa Barbara County and one site in Los Angeles County over multiple years. Adult I. pacificus became active in the winter (late November) and were rare or absent by late April to early May. Nymphal ticks became active in early to late February, were absent by early May to early June, and were rarely encountered using the drag method throughout their period of peak seasonal activity. Larval ticks became active earlier in the season, or at the same time as nymphs (early to late February) and were absent by early May. These results suggest a highly truncated period of I. pacificus seasonal questing activity, particularly apparent in the juvenile tick stages, in central and southern California relative to observed patterns in Lyme-endemic northwestern California. Notably, the highly truncated period of questing activity of the juvenile stages has important implications for pathogen transmission dynamics in that there exists only a brief window for horizontally transmitted pathogens to be acquired by one tick cohort and subsequently transmitted, through hosts, to the next tick cohort in this system. The broader patterns observed also suggest low human risk of tick-borne disease in central and southern California, and have implications for reduced tick-borne disease risk in the western US more generally under projected climate change.

  11. 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

  12. 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.

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

    SciTech Connect

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

    1992-05-01

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

  14. 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.

  15. Quantitative Immunofluorescent Blotting of the Multidrug Resistance-associated Protein 2 (MRP2)

    PubMed Central

    Gerk, Phillip M.

    2011-01-01

    Introduction Quantitation of the expression levels of proteins involved in drug transport and disposition is needed to overcome limitations of film-based detection of chemiluminescent immunoblots. Purpose The purpose was to describe and validate a quantitative immunofluorescent blotting method for detection of ATP-Binding Cassette Transporter Isoform C2/Multidrug Resistance-associated Protein 2 (ABCC2/MRP2). Methods Western blotting was performed by electrophoresis of membrane vesicle protein isolated from Sf9 cells overexpressing MRP2 subsequently blotting with infrared labeled secondary antibody. The bound complex was detected using the Odyssey Infrared Imaging System (Li-Cor; Lincoln, NE). The images were analyzed using the Odyssey Application Software to obtain the integrated intensities, followed by linear regression of the intensity data. Results The limits of quantitation for the time-insensitive technique described here were from 0.001μg to 0.5μg of total membrane protein, the coefficient of variation of the slope was 8.9%; r2 values were 0.986 ± 0.012. The utility and sensitivity of this technique was demonstrated in quantitating expression of MRP2 in human placental tissue samples, in which MRP2 was present in low abundance. Discussion The immunofluorescent blotting technique described provides sensitive, reproducible, and quantitative determinations of large, integral membrane proteins such as MRP2, all with potential long-term cost savings. PMID:21277982

  16. Mixed lubrication after rewetting of blotted pleural mesothelium.

    PubMed

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

    2013-01-15

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

  17. 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

  18. 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.

  19. 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.

  20. 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

  1. 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.

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

    USGS Publications Warehouse

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

    2009-01-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2013-04-01

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

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

    USGS Publications Warehouse

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

    1984-01-01

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

  5. 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

    2017-01-01

    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.

  6. 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.

  7. Vertical distribution of (137)Cs activity concentration in marine sediments at Amvrakikos Gulf, western of Greece.

    PubMed

    Tsabaris, C; Patiris, D L; Fillis-Tsirakis, E; Kapsimalis, V; Pilakouta, M; Pappa, F K; Vlastou, R

    2015-06-01

    The aim of the present work is the study of (137)Cs migration in sediment column taking into account the sedimentation rate in the Amvrakikos Gulf, at the western part of Greece. Marine core sediments were collected and the measurements were performed using the high resolution gamma-ray spectrometry method. The vertical distribution of (137)Cs activity concentration, as part of anthropogenic marine radioactivity, provided averaged sedimentation rate by identifying the depths of activity concentrations due to the Chernobyl accident and the nuclear tests signals. Furthermore, (137)Cs measurements were reproduced using the proposed one-dimensional diffusion-advection model which provides mainly as an output, the sedimentation rate and the average diffusivity of (137)Cs in the sediment column. The proposed model estimates the temporal variation of (137)Cs activity concentration from 1987 (one year after the Chernobyl accident) till today (2014).

  8. 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

  9. Multiplexed miRNA northern blots via hybridization chain reaction.

    PubMed

    Schwarzkopf, Maayan; Pierce, Niles A

    2016-09-06

    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.

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

    EPA Science Inventory

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

  11. 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.

  12. 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.

  13. 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.

  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. 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.

  16. Screening of Organophosphorus Acid Anhydrases from different Sources by Western Blot Analysis

    DTIC Science & Technology

    1991-01-01

    Glasel, J. A. et al., Mol. Immunol., Vol. 20, pp 1419-1422 (1983). 18. Kohen, F. et al., " Steriods ," Vol. 39, pp 453-459 (1982). 19. Bjercke, R. J. et al., J. Immunol. Meth. Vol. 90, pp 203-213 (1986) 20

  17. Indeterminate HIV-1 Western Blots: Etiology, Natural History, and Psychological Reactions

    DTIC Science & Technology

    1992-09-16

    homosexual men: decline before onset of illness related to acquired immune deficiency syndrome (AIDS). BMJ 1985;291:997-8. 16. Farzadegan H, Polls MA...with systemic lupus erythematous, Sjogren’s syndrome , and drug-induced autoimmunity. J Immunol 1986; 137: 25222-7. 33. Jack•on JB, Coombs RW...antibodies to retroviral proteins in parIents with primary Sjogren’s syndrome . Arthritis and Rheumatism 1990; 33:774-781. 51. Kunhl P, Seidl S, Holzberger

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

    PubMed

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

    2016-06-14

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

  19. 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

  20. Geodetic and geological evidence of active tectonics in south-western Sicily (Italy)

    NASA Astrophysics Data System (ADS)

    Barreca, G.; Bruno, V.; Cocorullo, C.; Cultrera, F.; Ferranti, L.; Guglielmino, F.; Guzzetta, L.; Mattia, M.; Monaco, C.; Pepe, F.

    2014-12-01

    Integrated geological, geodetic and marine geophysical data provide evidence of active deformation in south-western Sicily, in an area spatially coincident with the macroseismic zone of the destructive 1968 Belice earthquake sequence. Even though the sequence represents the strongest seismic event recorded in Western Sicily in historical times, focal solutions provided by different authors are inconclusive on possible faulting mechanism, which ranges from thrusting to transpression, and the seismogenic source is still undefined. Interferometric (DInSAR) observations reveal a differential ground motion on a SW-NE alignment between Campobello di Mazara and Castelvetrano (CCA), located just west of the maximum macroseismic sector. In addition, new GPS campaign-mode data acquired across the CCA alignment documents NW-SE contractional strain accumulation. Morphostructural analysis allowed to associate the alignment detected through geodetic measurements with a topographic offset of Pleistocene marine sediments. The on-land data were complemented by new high-resolution marine geophysical surveys, which indicate recent contraction on the offshore extension of the CCA alignment. The discovery of archaeological remains displaced by a thrust fault associated with the alignment provided the first likely surface evidence of coseismic and/or aseismic deformation related to a seismogenic source in the area. Results of the integrated study supports the contention that oblique thrusting and folding in response to NW-SE oriented contraction is still active. Although we are not able to associate the CCA alignment to the 1968 seismic sequence or to the historical earthquakes that destroyed the ancient Greek city of Selinunte, located on the nearby coastline, our result must be incorporated in the seismic hazard evaluation of this densely populated area of Sicily.

  1. 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

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

    NASA Astrophysics Data System (ADS)

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

    2003-12-01

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

  3. 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

  4. 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.

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

    PubMed

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

    2015-11-01

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

  6. 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.

  7. 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.

  8. 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.

  9. 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

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

    NASA Astrophysics Data System (ADS)

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

    2015-06-01

    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.

  12. Family Instability and Early Initiation of Sexual Activity in Western Kenya

    PubMed Central

    2014-01-01

    Epidemiological, economic, and social forces have produced high levels of volatility in family and household structure for young people growing up in sub-Saharan Africa in recent decades. However, scholarship on the family to date has not examined the influence of this family instability on young people’s well-being. The current study employs unique life history calendar data from Western Kenya to investigate the relationship between instability in caregiving and early initiation of sexual activity. It draws on a body of work on parental union instability in the United States, and examines new dimensions of family change. Analyses reveal a positive association between transitions in primary caregiver and the likelihood of early sexual debut that is rapidly manifested following caregiver change and persists for a short period. The association is strongest at early ages, and there is a cumulative effect of multiple caregiver changes. The results highlight the importance of studying family stability in sub-Saharan Africa, as distinct from family structure, and for attention to dimensions such as age and recency. PMID:23055236

  13. 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.

  14. Decadal change of Tropical Cyclone Activity over western North Pacific around late-1990s

    NASA Astrophysics Data System (ADS)

    He, H.; Yang, J.; Mao, R.; Wang, Y.; Gong, D.

    2014-12-01

    A pronounced decadal change of tropical cyclone (TC) activity was identified over western North Pacific (WNP) around late-1990s. After late-1990s, the WNP total TC genesis number exhibited an evident decrease, particularly over southern WNP region (S-WNP: 5oN-20oN), which was mainly caused by reduced vorticity and descending anomalies. We also detected a significant northward migration of TC genesis from 17.2°N to 18.7°N. The above TC genesis change is attributed to the weakening of monsoon trough and local Hadley cell that is associated with sea surface temperature climate shift around the late-1990s. In terms of three prevailing TC tracks changes, the northwestward-moving track (II) became the most dominant prevailing track mode while the westward-moving track (I) became weaker, and the northeastward-recurving track (III) had a westward shift. The track shifts primarily resulted from the large-scale steering flows change, which also had played a vital role in the modulation of TC regional duration. Thus, the subtropical East Asia tended to have a higher risk of encountering TC while the Southern China had a lower risk. Additionally, a visual reduction was seen in both number and proportion of typhoons reaching categories 1 and 2, and a remarkable poleward migration was also recognized in the average latitudes where TCs have achieved their lifetime-maximum intensity.

  15. 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

  16. 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

  17. 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.

  18. Applying phage antibodies to proteomics: selecting single chain Fv antibodies to antigens blotted on nitrocellulose.

    PubMed

    Liu, B; Marks, J D

    2000-11-01

    Two-dimensional gel electrophoresis is a powerful tool for identification of proteins that differ between patients with qualitatively or quantitatively different disease states. Further characterization of these protein differences would be greatly facilitated by the availability of antibodies that could be used to detect and quantitate the temporo-spatial pattern and cellular and tissue location of the different proteins. To generate such antibodies, methods were developed which permit the successful selection of monoclonal phage antibodies from phage display libraries against antigens blotted from SDS-PAGE gels onto nitrocellulose. First, it was determined that nitrocellulose and PVDF membranes gave significantly lower levels of background phage binding than two other membranes studied. Next, it was determined that blocking with fish gelatin and binding in the presence of 0.5 M NaCl could reduce nonspecific binding 10,000-fold and result in enrichment ratios greater than 500-fold with antigen concentrations as low as 1 ng/mm(2). When optimized conditions were applied to phage antibody libraries, panels of monoclonal phage antibodies were generated against the proteins ErbB2 and bovine serum albumin electroblotted from SDS-PAGE gels onto nitrocellulose. Antibodies were obtained with as little as 10 to 1 ng of antigen, depending on whether the libraries displayed single or multiple copies of antibody per phage. The antibodies worked as reagents in both ELISA and Western blotting.

  19. 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

  20. 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

  1. 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

  2. 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.

  3. 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

  4. Possible Linkage between Monsoon Trough Variability and Tropical Cyclone Activity over Western North Pacific: Role of Tropical Waves

    NASA Astrophysics Data System (ADS)

    Wu, Liang; Huang, Ronghui; Wen, Zhiping

    2014-05-01

    The present study investigates the influence of the monsoon trough (MT) on the interannual variability of tropical cyclone (TC) activity over the western North Pacific during July-November for the period 1979-2007. It is shown that the TC activity is closely related to the MT location. During the years when the MT extends eastward (retreats westward), more (less) TCs form within the southeastern quadrant of the western North Pacific. Such a relationship can be explained by the changes in tropical waves, such as mixed Rossby-gravity (MRG) waves and (tropical depression) TD-type disturbances, associated with the movement of the MT. An eastward extension of the MT coincides with enhanced TD-MRG type disturbances and a clear MRG-to-TD transition over the southeast quadrant of the western North Pacific. Such a transition is unclear during the years when the MT retreats westward. These waves associated with the eastern extension of the MT are favorable for TC genesis, while those associated with the westward retreat of the MT are not. Diagnosis of the barotropic energy conversion indicates that both the rotational and divergent components of the background flow change associated with MT are responsible for energy conversion from the mean flow to the TD-MRG perturbations. This is an important reason for the linkage between MT variability and TC genesis over the western North Pacific.

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

    DOE PAGES

    Carmichael, Joshua D.; Joughin, Ian; Behn, Mark D.; ...

    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

  6. Digestive organ sizes and enzyme activities of refueling western sandpipers (Calidris mauri): contrasting effects of season and age.

    PubMed

    Stein, R Will; Place, Allen R; Lacourse, Terri; Guglielmo, Christopher G; Williams, Tony D

    2005-01-01

    We examined seasonal and age-related variation in digestive organ sizes and enzyme activities in female western sandpipers (Calidris mauri) refueling at a coastal stopover site in southern British Columbia. Adult sandpipers exhibited seasonal variation in pancreatic and intestinal enzyme activities but not in digestive system or organ sizes. Spring migrants had 22% higher total and 67% higher standardized pancreatic lipase activities but 37% lower total pancreatic amylase activity than fall migrants, which suggests that the spring diet was enriched with lipids but low in glycogen. Spring migrants also had 47% higher total intestinal maltase activity as well as 56% higher standardized maltase and 13% higher standardized aminopeptidase-N activities. Spring migrants had higher total enzymic capacity than fall migrants, due primarily to higher total lipase and maltase activities. During fall migration, the juvenile's digestive system was 10% larger than the adult's, and it was composed differently: juveniles had a 16% larger small intestine but a 27% smaller proventriculus. The juvenile's larger digestive system was associated with lower total enzymic capacity than the adult's due to 20% lower total chitinase and 23% lower total lipase activities. These results suggest that juvenile western sandpipers may process food differently from adults and/or have a lower-quality diet.

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

    USGS Publications Warehouse

    Steven, Thomas A.

    1984-01-01

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

  8. 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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  10. 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

  11. Spectrophotometric quantitation of DNA on blots after ethanol-solubilization of the MTT-formazan from anti-digoxigenin-based detection of nucleic acids.

    PubMed

    Colgan, D J

    1993-01-01

    The tetrazolium salt, 3-(4,5-dimethyl-thiazolyl-2)-2,5-diphenyl tetrazolium bromide (MTT) has recently been established as a substitute for Nitro-blue tetrazolium (NBT) in stain mixtures using antibody-conjugated alkaline phosphatase for the location of proteins on Western blots (Heegaard, 1990). Experiments reported here show that MTT is as sensitive as NBT in digoxigenin-labeled probe localization (on nucleic acid blots) utilizing alkaline-phosphatase-labelled, anti-digoxigenin antibodies. Moreover, as the formazan from MTT is soluble in ethanol, it is shown that spectrophotometric quantitation can be used to estimate the amount of target DNA on dot and Southern blots. For dot blotting, pBR328 was used as the probe and pBR322 as target. For Southern blots, human rDNA was used as the probe and total genomic calf DNA as the target. Staining response was linear over at least six twofold DNA dilutions in both types of blot.

  12. 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.

  13. Volcanic activity recorded in deep-sea sediments and the geodynamic evolution of western Pacific island arcs

    NASA Astrophysics Data System (ADS)

    Cambray, Hervé; Pubellier, Manuel; Jolivet, Laurent; Pouclet, André

    A compilation of volcanic ashes interbedded in deep-sea sediments was carried out from DSDP-IPOD and ODP data collected along the western Pacific margin. Using a tephrochronological method, we attempted to reconstruct the Cenozoic and Quaternary volcanic activity of major western Pacific arcs. For every arc, established volcanic episodes and volcanic-tectonic evolution recorded on land were compared. This study reveals close connections between tectonic events and volcanic activity of arcs, as well a temporal relationship between the opening of marginal basins and arc volcanism. In the Tohoku (NE Japan) and Bonin arcs (SE Japan), arc volcanic activity clearly vanishes during backarc spreading. In contrast, intense volcanism occurs during both arc rifting and intervals of no spreading. Detailed comparisons show that the maximum volcanic output is closely connected with the stress field evolution recorded on land. The case of Seinan arc (SW Japan) shows a good fit between volcanic episodes and periods of release of the compressional stress field after major orogenic events. Furthermore, in the marine sediments off Japan, a systematic late Miocene volcanic hiatus interpreted as a quiescence of volcanic activity corresponds to a changing stress field on the Tohoku and Bonin arcs. These correlations between volcanic episodicity and tectonic evolution of island arcs allow us to discuss the influence of subduction process on arc volcanism. In the Philippines, the volcanic signal in marine sediments is compromised by rapid alteration and diagenesis of ashes. Nonetheless, only the main events of arc volcanic activity are preserved. A comparison with on land volcanism shows that this filtered volcanic signal in different places corresponds to incipient subduction (transition from passive to active margins) or to the final stages of basin closure.

  14. 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.

  15. 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.; 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.

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

    PubMed

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

    2010-04-01

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

  17. 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.

  18. 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).

  19. Antibacterial activity and chemical characteristics of several Western Australian honeys compared to manuka honey and pasture honey.

    PubMed

    Roshan, Niloufar; Rippers, Thomas; Locher, Cornelia; Hammer, Katherine A

    2017-03-01

    The physicochemical parameters and antibacterial activity of 10 Western Australian (WA) and two comparator honeys were determined. Honeys showed a pH range of 4.0-4.7, colour range of 41.3-470.7 mAU, methylglyoxal levels ranging from 82.2 to 325.9 mg kg(-1) and hydrogen peroxide levels after 2 h of 22.7-295.5 µM. Antibacterial activity was assessed by the disc diffusion assay, phenol equivalence assay, determination of minimum inhibitory and bactericidal concentrations and a time-kill assay. Activity was shown for all honeys by one or more method, however, activity varied according to which assay was used. Minimum inhibitory concentrations for WA honeys against 10 organisms ranged from 4.0 to >32.0% (w/v). Removal of hydrogen peroxide activity by catalase resulted in decreased activity for several honeys. Overall, the data showed that honeys in addition to those derived from Leptospermum spp. have antimicrobial activity and should not be overlooked as potential sources of clinically useful honey.

  20. Morphometric and meteorological controls on recent snow avalanche distribution and activity at hillslopes in steep mountain valleys in western Norway

    NASA Astrophysics Data System (ADS)

    Laute, Katja; Beylich, Achim A.

    2014-08-01

    Snow avalanches are common phenomena in Norway. Controlling factors of snow avalanche distribution and activity, and the relative importance of snow avalanches regarding contemporary sedimentary mass transfers were explored within two steep, parabolic-shaped and glacier-connected tributary valleys (Erdalen and Bødalen) in western Norway. Mapping of distribution, extent and the entire path lengths of snow avalanches was combined with spatial data analysis (GIS and DEM computing) of morphometric controls. The timing and frequency of snow avalanches were explored by correlating meteorological data with high-resolution monitoring data of snow avalanche events. Sediment masses annually transferred by snow avalanches along hillslopes and from hillslopes into stream channels were estimated. A high inter-annual variability of avalanche activity and a wide spectrum of avalanche sizes and types ranging from small to extreme-sized events were found for the four-year investigation period 2009-2012. Spatial distribution of snow avalanches is governed by the topographical factors valley orientation, slope aspect, relative slope height and rockwall morphometry whereas timing and frequency of snow avalanches are controlled by snowfall intensity, periods with strong winds combined with a prevalent wind direction or sharp air temperature changes within short time periods. Snow avalanches represent one of the dominant denudational processes and have a high relative importance regarding sedimentary mass transfers within the two mountain valleys Erdalen and Bødalen in western Norway.

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

    PubMed

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

    2016-02-18

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

  2. 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

  3. 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

  4. Discovery of a novel insecticidal protein from Chromobacterium piscinae, with activity against Western Corn Rootworm, Diabrotica virgifera virgifera.

    PubMed

    Sampson, Kimberly; Zaitseva, Jelena; Stauffer, Maria; Vande Berg, Brian; Guo, Rong; Tomso, Daniel; McNulty, Brian; Desai, Nalini; Balasubramanian, Deepa

    2017-01-01

    Western corn rootworm (WCR), Diabrotica virgifera virgifera, is one of the most significant pests of corn in the United States. Although transgenic solutions exist, increasing resistance concerns make the discovery of novel solutions essential. In order to find a novel protein with high activity and a new mode of action, a large microbial collection was surveyed for toxicity to WCR using in vitro bioassays. Cultures of strain ATX2024, identified as Chromobacterium piscinae, had very high activity against WCR larvae. The biological activity from the strain was purified using chromatographic techniques and fractions were tested against WCR larvae. Proteins in the final active fraction were identified by mass spectrometry and N-terminal sequencing and matched to the genome of ATX2024. A novel 58.9kDa protein, identified by this approach, was expressed in a recombinant expression system and found to have specific activity against WCR. Transgenic corn events containing this gene showed good protection against root damage by WCR, with average scores ranging between 0.01 and 0.04 on the Iowa State node injury scale. Sequence analysis did not reveal homology to any known insecticidal toxin, suggesting that this protein may act in a novel way to control WCR. The new WCR active protein is named GNIP1Aa, for Gram Negative Insecticidal Protein.

  5. 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.

  6. 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

  7. 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

  8. 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.

  9. Significant difference in active metabolite levels of ginseng in humans consuming Asian or Western diet: The link with enteric microbiota.

    PubMed

    Wan, Jin-Yi; Wang, Chong-Zhi; Zhang, Qi-Hui; Liu, Zhi; Musch, Mark W; Bissonnette, Marc; Chang, Eugene B; Li, Ping; Qi, Lian-Wen; Yuan, Chun-Su

    2017-04-01

    After ingestion of ginseng, the bioavailability of its parent compounds is low and enteric microbiota plays an important role in parent compound biotransformation to their metabolites. Diet type can influence the enteric microbiota profile. When human subjects on different diets ingest ginseng, their different gut microbiota profiles may influence the metabolism of ginseng parent compounds. In this study, the effects of different diet type on gut microbiota metabolism of American ginseng saponins were investigated. We recruited six healthy adults who regularly consumed different diet types. These subjects received 7 days' oral American ginseng, and their biological samples were collected for LC-Q-TOF-MS analysis. We observed significant ginsenoside Rb1 (a major parent compound) and compound K (a major active metabolite) level differences in the samples from the subjects consuming different diets. Subjects on an Asian diet had much higher Rb1 levels but much lower compound K levels compared with those on a Western diet. Since compound K possesses much better cancer chemoprevention potential, our data suggested that consumers on a Western diet should obtain better cancer prevention effects with American ginseng intake compared with those on an Asian diet. Ginseng compound levels could be enhanced or reduced via gut microbiota manipulation for clinical utility.

  10. [Sanitary and relief activity of the Polish Red Cross in the repatriation and displacement action in the Western Pomeranian region].

    PubMed

    Kedzia, A

    2001-01-01

    The final period of World War II and the first months of peace were the time of massive migration of Polish and German population. The first stage of the Poles' repatriation from the territories taken over by the USSR to the liberated parts of Poland underwent in an organizational chaos, which resulted from poor coordination between the institutions responsible for this operation. In principle, the National Repatriation Office was in charge of the repatriation action. However, where the NRO had no branches or wanted a joint action, the Polish Red Cross took a vital part in sanitary and relief activities. They were particularly crucial in the western and northern regions of Poland, which faced three massive migration currents: the influx of Polish repatriates from the USSR, the return of concentration camp prisoners, POWs and slave workers from Germany and the displacement of the German population. Colonel Jan Ciałowicz gave a brief résumé of the role the Polish Red Cross played in the repatriation and displacement action: "... At the time when the foundations of the State of Poland were created in the Western Pomeranian Region, the Polish Red Cross was dressing, feeding and treating those who needed it most desperately, taking special care of the repatriates and children."

  11. 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.

  12. Antimicrobial activity of crude fractions and morel compounds from wild edible mushrooms of North western Himalaya.

    PubMed

    Shameem, Nowsheen; Kamili, Azra N; Ahmad, Mushtaq; Masoodi, F A; Parray, Javid A

    2017-04-01

    The antimicrobial properties of morel compounds from wild edible mushrooms (Morchella esculenta and Verpa bohemica) from Kashmir valley was investigated against different clinical pathogens. The butanol crude fraction of most popular or true morel M. esculenta showed highest 19 mm IZD against E.coli while as same fraction of Verpa bohemica exhibited 15 mm IZD against same strain. The ethyl acetate and butanol crude fractions of both morels also exhibited good antifungal activity with highest IZD shown against A. fumigates. The three morel compounds showed quite impressive anti bacterial and fungal activities. The Cpd 3 showed highest inhibitory activity almost equivalent to the synthetic antibiotics used as control. The MIC/MBC values revealed the efficiency of isolated compounds against the pathogenic strains. In the current study significant inhibitory activity of morel compounds have been obtained paying the way for their local use from ancient times.

  13. 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)

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

    PubMed

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

    2014-05-01

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

  15. TESA-blot for the diagnosis of Chagas disease in dogs from co-endemic regions for Trypanosoma cruzi, Trypanosoma evansi and Leishmania chagasi.

    PubMed

    Umezawa, E S; Souza, A I; Pinedo-Cancino, V; Marcondes, M; Marcili, A; Camargo, L M A; Camacho, A A; Stolf, A M S; Teixeira, M M G

    2009-07-01

    We standardized serodiagnosis of dogs infected with Trypanosoma cruzi using TESA (trypomastigote excreted-secreted antigen)-blot developed for human Chagas disease. TESA-blot showed 100% sensitivity and specificity. In contrast, ELISA using TESA (TESA-ELISA) or epimastigotes (epi-ELISA) as antigen yielded 100% sensitivity but specificity of 94.1% and 49.4%, respectively. When used in field studies in an endemic region for Chagas disease, visceral leishmaniasis and Trypanosoma evansi (Mato Grosso do Sul state, Central Brazil), positivities were 9.3% for TESA-blot, 10.7% for TESA-ELISA and 32% for epi-ELISA. Dogs from a non-endemic region for these infections (Rondonia state, western Amazonia) where T. cruzi is enzootic showed positivity of 4.5% for TESA-blot and epi-ELISA and 6.8% for TESA-ELISA. Sera from urban dogs from Santos, São Paulo, where these diseases are absent, yielded negative results. TESA-blot was the only method that distinguished dogs infected with T. cruzi from those infected with Leishmania chagasi and/or Trypanosoma evansi.

  16. Retraction: Characterization of cellulolytic activities of newly isolated Thelephora sowerbyi from North-Western Himalayas on different lignocellulosic substrates.

    PubMed

    Sharma, Deepika; Goel, Gunjan; Bansal, Saurabh; Mahajan, Rishi; Sharma, B M; Chauhan, Rajinder Singh

    2016-12-01

    Characterization of cellulolytic activities of newly isolated Thelephora sowerbyi from North-Western Himalayas on different lignocellulosic substrate J. Basic Microbiol. 2015, 55, 1-11 - DOI: 10.1002/jobm.201500107 The above article from the Journal of Basic Microbiology, published online on 08 June 2015 in Wiley Online Library as Early View (http://onlinelibrary.wiley.com/doi/10.1002/jobm.201500107/pdf), has been retracted by agreement between the authors, the Editor-in-Chief and Wiley-VCH GmbH & Co. KGaA. The retraction has been agreed because the microorganism studied in the described experiments has been identified as the fungus Cotylidia pannosa (Gene Accession No. KT008117) instead of Thelephora sowerbyi. The culture has been identified on the basis of the sequence of the amplified ITS region of the microorganism which was submitted by the authors to the NCBI database.

  17. A 20th century calendar of snow avalanche activity within the Bødalen valley, inner Nordfjord, western Norway

    NASA Astrophysics Data System (ADS)

    Decaulne, A.; Eggertsson, O.; Laute, K.; Beylich, A. A.

    2012-04-01

    The forest-covered valley floor of the Bødalen valley located in the inner Nordfjord, western Norway offers the great opportunity for snow-avalanche reconstruction using tree rings. By sampling at different points within the forest stand, both on birch (Betula pubescens) and alder (Alnus), we are able to extract the magnitude and return period of major snow-avalanche events in the valley. The years of 1967, 1974, 1985, 1994 and 2001 appear like very avalanche-rich, and the years of 1992 and 2007 appear as extreme, with snow-avalanche signals being recorded at the majority of the trees. Although the investigated valley is quite remote during the winter time, the gained knowledge of the recent snow-avalanche history provides a picture of the potential activity that can be expected also elsewhere in the Nordfjord region, and escpecially in the inhabited areas as well as along important transportation corridors that do not have such records available.

  18. Effects of Chinese, Japanese and Western tea on hepatic P450 enzyme activities in rats.

    PubMed

    Niwattisaiwong, N; Luo, X X; Coville, P F; Wanwimolruk, S

    2004-01-01

    Previous studies have reported that green tea effectively protects against cancers caused by various dietary carcinogens. As P450 enzymes are the major system responsible for the metabolism of many carcinogens, we hypothesise that tea consumption may alter the catalytic activities of P450 enzymes. We conducted this study to screen the effects of four different teas on the activities of P450 enzymes. Tea solutions (2.5%) were prepared by adding boiling water to tea leaves and filtering. Female Wistar rats were divided into five groups (n = 4 each); each had free access to tea solutions while the control group was supplied with water for 4 weeks. Animals were sacrificed and livers were removed for preparation of microsomes. Enzyme activities were determined by incubation of liver microsomes with the appropriate CYP substrate. The activity of CYP1A1 in livers from rats receiving Oolong (Chinese) tea (185 +/- 63 pmol/mg/min), Japanese green tea (197 +/- 22 pmol/mg/min) and Earl Grey tea (228 +/- 40 pmol/mg/min) was significantly higher (p < 0.05) than in the control group (94 +/- 34 pmol/mg/min), whereas no change was observed in the activity of CYP1A2 in any of tested animals. The hepatic activity of CYP2D6 was greater only in rats drinking Earl Grey tea compared to the controls (235 +/- 37 vs 161 +/- 41 pmol/mg/min, p < 0.05). There were also significant increases (p < 0.05) in the activity of CYP3A in livers of animals given Oolong tea (653 +/- 174 vs 382 +/- 114 pmol/mg/min) and Earl Grey tea (751 +/- 202 pmol/mg/min), while Jasmine and Japanese green tea had no significant effect. These results indicate that not all types of tea cause alterations in liver CYP enzymes as some elevated activities and some did not. Further studies are needed to determine whether there is a relationship between the effect of tea on CYP activities and anti-carcinogenesis.

  19. 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.

  20. 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.

  1. Recent seismic activity of the Kivu Province, Western Rift Valley of Africa

    NASA Astrophysics Data System (ADS)

    Zana, N.; Kamba, M.; Katsongo, S.; Janssen, Th.

    1989-11-01

    The Kivu Province is located at the junction between the well-defined Ruzizi Valley to the south and the Lake Amin Trough to the north. In this zone, the Rift Valley is characterized by the highest uplift and by complex dislocations of the crust, accompanied by the most intensive volcanism of the East African Rift System. In this paper, we show the recent state of the seismic activity of this zone in connection with the seismic activity generated by the volcanoes Nyiragongo and Nyamuragira. The pattern of cumulative energy release by these volcanoes shows a steplike increase that is believed to be a precursor of volcanic eruptions.

  2. Parafilm-M®, An Available Cost-Effective Alternative for Immuno-blot Pouches.

    PubMed

    Quadri, Syed M S

    2015-01-01

    Commercially available standard immuno-blot pouches do play an efficient role in antibody incubation in performing an immuno-blot, but are not readily available in the laboratory and have to be specifically ordered. We have developed an equally efficient technique to make an immune-blot more cost-effective with more conservation of antibodies by using a common and readily available laboratory product Parafilm-M(®). Parafilm-M(®) which serves as a sealant for various items of laboratory equipment can be used for antibody incubation. Manually made Parafilm-M(®) pouch has a clear advantage over standard immuno-blot pouches in terms of availability, cost-effectiveness, and consumption of antibodies that ultimately reduces the cost of an immuno-blot. We have performed a series of experiments to check the efficacy of both the techniques. Samples with equal amount of protein were analyzed on separate SDS PAGE gels. The proteins were transferred electrophoretically to the nitrocellulose membrane using Trans-Blot(®) Turbo™ Mini Nitrocellulose Transfer Pack. Antibody incubation was done using standard immuno-blot pouch, standard container and Parafilm-M(®) sealed pouch. The expression of protein was determined and the results of immuno-blots were compared. We found that antibodies are binding the membrane in Parafilm-M(®) pouches as efficiently as in container method or in standard immuno-blot pouches. By restricting the membrane, the surface area of the manually made Parafilm-M(®) pouch can be reduced, less diluent is required to cover the membrane as a result less antibodies are consumed. We also calculated that each immuno-blot pouch cost around $0.1906, whereas the cost for Parafilm-M(®) pouch is 0.0695 which is almost one-third the price of an immuno-blot pouch. Thus, Parafilm-M(®) method distinctly provides a cost-effective solution for antibody incubation.

  3. 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.

  4. 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…

  5. Constraints on Alpine hydrothermal activity and deformation from U-Th-Pb dating of cleft monazite and xenotime (Western Alps)

    NASA Astrophysics Data System (ADS)

    Grand'Homme, Alexis; Janots, Emilie; Bosse, Valerie; Seydoux-Guillaume, Anne-Magali; De Ascencao, Roger

    2016-04-01

    In this large-scale regional study, age of hydrothermal monazite (and xenotime) precipitation has been investigated through in-situ U-Th-Pb dating of crystals collected in 11 clefts (veins) taken in the internal and external massifs (Western Alps). The investigated clefts are composed of quartz, chlorite (± epidote), albite and millimetric accessory minerals (monazite, apatite, xenotime, anatase, rutile). Prior to dating, cleft monazite composition was thoroughly studied to reveal potential zoning. In-situ dating through different compositional domains of single monazite crystal yields well-resolved Th-Pb ages (typically with 0.1-0.3 Ma resolution) indicating for growth episodes with short duration. Comparison of U-Pb and Th-Pb dating indicates that the U-Pb systematics appears successful to date cleft monazite with low Th/U ratio (typically <30). In one cleft, in which monazite and xenotime coexist, xenotime was dated using the monazite analytical protocol. Hydrothermal xenotime has remarkably high Th/U ratio and U-Pb dating shows evidence of 206Pb excess. In comparison, Th-Pb dating gives robust ages (35.2 ± 0.8 Ma) that are close but higher than the monazite date obtained in the same cleft (32.3 ± 0.3 Ma). Brief episodic monazite crystallization is likely attributed to enhanced hydrothermal activity during periods of higher tectonic activity. Correlation with other geochronological data suggests that it occurs in a host-rock that already cooled down at temperature close or below to the zircon fission track. In the Belledonne massif, the new monazite ages confirm for two periods of hydrothermal activity at around 11-13 Ma and 8-6 Ma, respectively. Only one vertical cleft monazite was investigated in the Mont-Blanc massif but it gives an age that is similar to the early population of the Belledonne massif (11.1 ± 0.2 Ma). Monazite dating therefore suggests for similar late-stage tectonic activity from Belledonne up to the Aar massifs, likely due to dextral

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

    NASA Astrophysics Data System (ADS)

    Arendse, Gillian J.

    2009-04-01

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

  7. 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.

  8. 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

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

    PubMed

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

    2014-10-01

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

  10. Distribution and activity of ice wedges across the forest-tundra transition, western Arctic Canada

    NASA Astrophysics Data System (ADS)

    Kokelj, S. V.; Lantz, T. C.; Wolfe, S. A.; Kanigan, J. C.; Morse, P. D.; Coutts, R.; Molina-Giraldo, N.; Burn, C. R.

    2014-09-01

    Remote sensing, regional ground temperature and ground ice observations, and numerical simulation were used to investigate the size, distribution, and activity of ice wedges in fine-grained mineral and organic soils across the forest-tundra transition in uplands east of the Mackenzie Delta. In the northernmost dwarf-shrub tundra, ice wedge polygons cover up to 40% of the ground surface, with the wedges commonly exceeding 3 m in width. The largest ice wedges are in peatlands where thermal contraction cracking occurs more frequently than in nearby hummocky terrain with fine-grained soils. There are fewer ice wedges, rarely exceeding 2 m in width, in uplands to the south and none have been found in mineral soils of the tall-shrub tundra, although active ice wedges are found there throughout peatlands. In the spruce forest zone, small, relict ice wedges are restricted to peatlands. At tundra sites, winter temperatures at the top of permafrost are lower in organic than mineral soils because of the shallow permafrost table, occurrence of phase change at 0°C, and the relatively high thermal conductivity of icy peat. Due to these factors and the high coefficient of thermal contraction of frozen saturated peat, ice wedge cracking and growth is more common in peatlands than in mineral soil. However, the high latent heat content of saturated organic active layer soils may inhibit freezeback, particularly where thick snow accumulates, making the permafrost and the ice wedges in spruce forest polygonal peatlands susceptible to degradation following alteration of drainage or climate warming.

  11. 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

  12. 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.

  13. 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

  14. 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...

  15. 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...

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

    PubMed

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

    2014-06-01

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

  17. Incidence and diagnosis of active toxoplasma infection among liver transplant recipients in Western Turkey.

    PubMed

    Caner, Ayşe; Döşkaya, Mert; Karasu, Zeki; Değirmenci, Aysu; Guy, Edward; Kiliç, Murat; Zeytunlu, Murat; Francis, Janet; Bozoklar, Ata; Gürüz, Yüksel

    2008-10-01

    Toxoplasmosis is a serious and potentially life-threatening disease in liver transplant recipients while they are immunosuppressed. We report the clinical and laboratory findings related to active toxoplasma infection associated with 40 immunosuppressed liver transplant procedures that took place over a 12-month period at a major transplant unit in Izmir, Turkey. Twenty-seven (67.5%) of the 40 transplant recipients were found to be seropositive for toxoplasma infection and therefore at risk of reactivated infection. From the serological status of the donors, which was ascertained in 38 of 40 cases, we identified 3 (7.9%) of 38 transplants to be from a seropositive donor to a seronegative recipient. In 10 (26.3%) of 38 transplants, both the donor and recipient were seronegative, and this excluded toxoplasma as a risk. A comparison of real-time polymerase chain reaction (PCR) and nested PCR was undertaken in combination with a range of serological assays (the Sabin-Feldman dye test, enzyme immunoassay immunoglobulin M, and immunosorbent agglutination assay immunoglobulin M). Ethylene diamine tetraacetic acid blood samples from 3 of the 30 recipients at risk from toxoplasma were found positive by PCR, but only 1 of these was found positive in both assays. Among the 3 PCR-positive patients, immunoglobulin M and immunoglobulin G antibody levels increased in only 1 patient. Correlations between symptoms, laboratory findings, and clinical management (use of anti-toxoplasma therapy) are presented. Our findings suggest that toxoplasma presents a significant risk to our liver transplant population and that PCR is a helpful addition in identifying active infections and hence in informing clinical management decisions.

  18. 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.

  19. 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.

  20. 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

  1. An investigation of reduced western disturbance activity over Northwest India in November - December 2015 compared to 2014 - A case study

    NASA Astrophysics Data System (ADS)

    Basu, Soumik; Bieniek, Peter A.; Deoras, Akshay

    2017-02-01

    In November-December of 2015, Northwestern India received very low precipitation due to anomalously low Western Disturbances (WDs) activity. The resulting lack of sufficient precipitation and soil moisture hampered the growth of winter crops leading to significant agricultural losses. Relatively stable weather in the absence of precipitation and WDs contributed to extremely high air pollution in New Delhi and also significantly degraded the air quality in many cities of Northwestern India leading to severe health issues. Despite the fact that WDs play a very important role in India's winter weather, limited research has been done to investigate the causes of their inter-annual variability. A case study using NCEP/NCAR Reanalysis, CMAP precipitation and NOAA Extended Reconstructed Sea Surface Temperature data is evaluated in this paper to better understand the atmospheric drivers of WDs in order to help fill the gap in knowledge. Results show that elevated Sea Surface Temperatures over the North Indian Ocean likely lead to atmospheric circulation anomalies that led to branching and weakening of the subtropical jet stream and weakening of vertical wind shear over Northwestern India. These conditions created an unfavorable environment for the propagation of WDs. However, there was an intensification of vertical wind shear over mid-latitude Eurasia along with increased storm activity. This weakened the Eurasian anticyclone resulting in warmer surface air temperatures over the midlatitudes that led to a redistribution of the meridional temperature gradient.

  2. 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

  3. 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.

  4. Active Tectonics of Western Turkmenistan; Implications for the Onset of South Caspian Subduction

    NASA Astrophysics Data System (ADS)

    Hollingsworth, J.; Jackson, J.; Priestley, K.

    2007-12-01

    The Kopeh Dagh and Balkan mountain ranges of West Turkmenistan are actively deforming as a result of Arabia- Eurasia collision. We combine observations of the geomorphology made from satellite and topographic data, with historical and recent seismicity to identify major active faults, and how they contribute to regional shortening. Between 55--57.5°E, partitioned (north-vergent) thrust and right-lateral strike-slip fault segments, comprising the Ashkabad fault zone, accommodate regional shortening and the westward-extrusion of the NW Kopeh Dagh-South Caspian block, relative to Central Iran and Eurasia. Reconstruction of displaced geology indicates 35~km total right-lateral motion across the Ashkabad fault zone. The Balkan region lies along-strike of the Ashkabad fault zone, west of 55°E. Fault plane solutions indicate shortening is partitioned onto the Balkan thrust and right-lateral Kum-Dagh fault zones. Thrust earthquakes are relatively deep (30--45~km) and lie along a north-dipping plane which extends 40±5~km north beneath the Balkan anticline. Receiver function data from Turkmenbashi and Nebit Dagh indicate these earthquakes occur in the base of the crust, and may therefore be related to bending of the NW Kopeh Dagh-South Caspian lithosphere as it is overthrust by Eurasia. Movement on a north-dipping blind thrust fault is consistent with the broad asymmetric (south-vergent) fold structure of the Balkan range. Recent uplift is also indicated by extensional faults which displace Quaternary geomorphology along the range crest. South of the Balkan range, right-lateral shear occurs across the Kum-Dagh fault zone which is expressed as a series of right-stepping anticlines (affecting Pliocene Red Series and younger sediments), forming important traps for hydrocarbons. An important structural change occurs near 55°E. To the west, Eurasia overthrusts the NW Kopeh Dagh- South Caspian block, while to the east the polarity of thrusting changes and the Kopeh Dagh

  5. Introduced annual grass increases regional fire activity across the arid western USA (1980-2009).

    PubMed

    Balch, Jennifer K; Bradley, Bethany A; D'Antonio, Carla M; Gómez-Dans, José

    2013-01-01

    Non-native, invasive grasses have been linked to altered grass-fire cycles worldwide. Although a few studies have quantified resulting changes in fire activity at local scales, and many have speculated about larger scales, regional alterations to fire regimes remain poorly documented. We assessed the influence of large-scale Bromus tectorum (hereafter cheatgrass) invasion on fire size, duration, spread rate, and interannual variability in comparison to other prominent land cover classes across the Great Basin, USA. We compared regional land cover maps to burned area measured using the Moderate Resolution Imaging Spectroradiometer (MODIS) for 2000-2009 and to fire extents recorded by the USGS registry of fires from 1980 to 2009. Cheatgrass dominates at least 6% of the central Great Basin (650 000 km(2) ). MODIS records show that 13% of these cheatgrass-dominated lands burned, resulting in a fire return interval of 78 years for any given location within cheatgrass. This proportion was more than double the amount burned across all other vegetation types (range: 0.5-6% burned). During the 1990s, this difference was even more extreme, with cheatgrass burning nearly four times more frequently than any native vegetation type (16% of cheatgrass burned compared to 1-5% of native vegetation). Cheatgrass was also disproportionately represented in the largest fires, comprising 24% of the land area of the 50 largest fires recorded by MODIS during the 2000s. Furthermore, multi-date fires that burned across multiple vegetation types were significantly more likely to have started in cheatgrass. Finally, cheatgrass fires showed a strong interannual response to wet years, a trend only weakly observed in native vegetation types. These results demonstrate that cheatgrass invasion has substantially altered the regional fire regime. Although this result has been suspected by managers for decades, this study is the first to document recent cheatgrass-driven fire regimes at a regional

  6. GTP-blot analysis of small GTP-binding proteins. The C-terminus is involved in renaturation of blotted proteins.

    PubMed

    Klinz, F J

    1994-10-01

    Recombinant c-Ha-ras, ralA and rap2, but not rap1A or rap1B proteins retained their ability to bind [alpha-32P]GTP after SDS/PAGE and transfer to nitrocellulose. Recombinant c-Has-ras missing the C-terminal 23 amino acid residues failed to bind [alpha-32P]GTP after the blot, and the ability of recombinant ralA missing the C-terminal 28 amino acid residues to bind [alpha-32P]GTP was decreased many-fold. The presence of nonionic detergents of the polyoxyethylene type such as Tween 20, Triton X-100, Nonidet P40 or Lubrol PX in the incubation buffer was necessary to induce renaturation of blotted recombinant c-Ha-ras protein, whereas other types of detergents were ineffective. We propose that detergents of the polyoxyethylene type induce the refolding of some types of blotted small GTP-binding proteins and that the C-terminus is involved in the refolding process. Membranes from NIH3T3 fibroblasts overexpressing c-Ha-ras protein showed much weaker binding of [alpha-32P]GTP as expected from the level of ras immunoreactivity. Treatment of fibroblasts with lovastatin, an inhibitor of hydroxymethylglutaryl-coenzyme A reductase, caused the accumulation of the unfarnesylated form of c-Ha-ras in the cytosol. Examination of [alpha-32P]GTP-binding and immunoreactivity for cytosolic and membrane-bound c-Ha-ras revealed that binding of [alpha-32P]GTP to unprocessed c-Ha-ras was increased about threefold compared to the same amount of processed c-Ha-ras. Our results demonstrate that detection and quantification of small GTP-binding proteins in eukaryotic cells by GTP-blot analysis is hampered by the fact that these proteins differ strongly in their ability to renature after blotting to nitrocellulose.

  7. Detection and quantitation of HPV DNA replication by Southern blotting and real-time PCR.

    PubMed

    Morgan, Iain M; Taylor, Ewan R

    2005-01-01

    This provides a brief introduction into the mechanism of DNA replication by the E1 and E2 proteins and describes the traditional Southern blotting technique that is used to monitor E1- and E2-mediated DNA replication. It also includes a novel real-time polymerase chain reaction (PCR) approach for monitoring E1- and E2-mediated DNA replication that has enhanced sensitivity and quantitation compared with Southern blotting, and a discussion of when to use the Southern blotting and real-time PCR techniques.

  8. 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

  9. Morphometric and meteorological controls of snow avalanche distribution and activity at hillslopes in steep mountain valleys in western Norway

    NASA Astrophysics Data System (ADS)

    Laute, Katja; Beylich, Achim A.

    2013-04-01

    Snow avalanches are common phenomena in Norway due to the interactions between the prevalent climatic factors and local topography. Research on snow avalanches provides insights into possible effects of predicted climate change on avalanche activity and connected sediment transport in mountain areas. This study focuses on (i) controlling factors of avalanche distribution and activity, and (ii) their relative importance regarding mass transfers in two steep, parabolic-shaped and glacier-connected tributary valleys (Erdalen and Bødalen) in western Norway. Mapping of distribution, extension and run-out distances of avalanches is combined with spatial data analysis of morphometric controls. Based on correlation of climate data with monitored avalanche events the timing and frequency of avalanches is explored and debris mass transfer on hillslopes caused by avalanches is estimated. The denudative effect of snow avalanches occurs in two steps: firstly throughout erosion directly on the surface of the rockwall and secondly due to their transport ability which causes significant remobilization and transport of available debris further downslope. The spatial distribution of snow avalanches depends on the valley orientation, slope aspect and rockwall morphometry. Especially distinct laterally convex-shaped leeside upper rockwall areas allow a high accumulation rate of snow during winter which is then released as avalanches during spring. The timing and frequency of avalanches in both valleys depend mainly on snowfall intensity, periods with strong winds combined with a stable wind direction or sudden air temperature changes. Snow avalanche activity leads in some valley areas to significant hillslope-channel coupling because debris is transported far enough by avalanches to reach channels. Snow avalanches represent one of the dominant denudational processes and have a high relative importance regarding mass transfer within the sedimentary budgets of the entire valleys.

  10. 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).

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

    PubMed

    Ranganathan, Srinivasan; Swaminathan, Subramanian

    2015-04-01

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

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

    PubMed

    Ranganathan, Srinivasan; Swaminathan, Subramanian

    2015-03-31

    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.

  13. 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

  14. 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

  15. 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.

  16. 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.

  17. 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

    2015-01-07

    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.

  18. 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.

  19. 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.

  20. 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

  1. 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 Ho

    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.

  2. Progress, opportunities, and key fields for groundwater quality research under the impacts of human activities in China with a special focus on western China.

    PubMed

    Li, Peiyue; Tian, Rui; Xue, Chenyang; Wu, Jianhua

    2017-03-10

    Groundwater quality research is extremely important for supporting the safety of the water supply and human health in arid and semi-arid areas of China. This review article was constructed to report the latest research progress of groundwater quality in western China where groundwater quality is undergoing fast deterioration because of fast economic development and extensive anthropogenic activities. The opportunities brought by increasing public awareness of groundwater quality protection were also highlighted and discussed. To guide and promote further development of groundwater quality research in China, especially in western China, ten key groundwater quality research fields were proposed. The review shows that the intensification of human activities and the associated impacts on groundwater quality in China, especially in western China, has made groundwater quality research increasingly important, and has caught the attention of local, national, and international agencies and scholars. China has achieved some progress in groundwater quality research in terms of national and regional laws, regulations, and financial supports. The future of groundwater quality research in China, especially in western China, is promising reflected by the opportunities highlighted. The key research fields proposed in this article may also inform groundwater quality protection and management at the national and international level.

  3. Blot-based detection of dehydroalanine-containing glutathione peroxidase with the use of biotin-conjugated cysteamine.

    PubMed

    Rhee, Sue Goo; Cho, Chun-Seok

    2010-01-01

    Dehydroalanine (DHA), alpha,beta-unsaturated amino acid, is found in the position corresponding to the serine, cysteine, and selenocysteine (Sec) residues of various proteins. Proteinaceous Sec is readily oxidized and subsequently undergoes beta-elimination to produce DHA. Glutathione peroxidase (GPx), which contains a Sec at the active site, is irreversibly inactivated by its own substrate as the result of the oxidation of selenium atom followed by the conversion of oxidized Sec to DHA. We developed a convenient method for estimation of the amount of DHA-GPx1 in cell homogenates. This blot-based method depends on specific addition of biotin-conjugated cysteamine to the DHA residue followed by detection of biotinylated protein based on its interaction with streptavidin. The method required an immunoprecipitation of GPx1 before labeling with the cysteamine derivative because many other proteins contain DHA. With the use of this method, we found that conversion of the Sec residue at the active site of GPx1 to DHA occurred during aging of red blood cells (RBCs) in vivo as well as in RBCs exposed to H(2)O(2) generated either externally by glucose oxidase or internally as a result of aniline-induced Hb autoxidation. Accordingly, the content of DHA-GPx1 in each RBC likely reflects total oxidative stress experienced by the cell during its lifetime of 120 days. Previous studies suggested that the activity of GPx1 in RBCs is most influenced by lifestyle and environmental factors such as the use of dietary supplements and smoking habit. Therefore, DHA-GPx1 in RBCs might be a suitable surrogate marker for evaluation of oxidative stress in the body. Our blot-based method for the detection of DHA-GPx1 will be very useful for evaluation of such stress. In addition, similar blot detection method can be devised for other proteins for which immunoprecipitating antibodies are available.

  4. Abundance and single-cell activity of heterotrophic bacterial groups in the western Arctic Ocean in summer and winter.

    PubMed

    Nikrad, Mrinalini P; Cottrell, M T; Kirchman, D L

    2012-04-01

    Environmental conditions in the western Arctic Ocean range from constant light and nutrient depletion in summer to complete darkness and sea ice cover in winter. This seasonal environmental variation is likely to have an effect on the use of dissolved organic matter (DOM) by heterotrophic bacteria in surface water. However, this effect is not well studied and we know little about the activity of specific bacterial clades in the surface oceans. The use of DOM by three bacterial subgroups in both winter and summer was examined by microautoradiography combined with fluorescence in situ hybridization. We found selective use of substrates by these groups, although the abundances of Ant4D3 (Antarctic Gammaproteobacteria), Polaribacter (Bacteroidetes), and SAR11 (Alphaproteobacteria) were not different between summer and winter in the Beaufort and Chukchi Seas. The number of cells taking up glucose within all three bacterial groups decreased significantly from summer to winter, while the percentage of cells using leucine did not show a clear pattern between seasons. The uptake of the amino acid mix increased substantially from summer to winter by the Ant4D3 group, although such a large increase in uptake was not seen for the other two groups. Use of glucose by bacteria, but not use of leucine or the amino acid mix, related strongly to inorganic nutrients, chlorophyll a, and other environmental factors. Our results suggest a switch in use of dissolved organic substrates from summer to winter and that the three phylogenetic subgroups examined fill different niches in DOM use in the two seasons.

  5. 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.

  6. 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

  7. 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.

  8. 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

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

    NASA Astrophysics Data System (ADS)

    Chang, C.

    2013-05-01

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

  10. 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

  11. Seasonal Modulations of Different Impacts of Two Types of ENSO Events on Tropical Cyclone Activity in the Western North Pacific

    NASA Astrophysics Data System (ADS)

    LI, C.; Wang, C.

    2013-12-01

    The paper examines different impacts of eastern Pacific warm/cold (EPW/EPC) and central Pacific warm/cold (CPW/CPC) events on tropical cyclones (TCs) in the western North Pacific (WNP) by considering the early season of April-June (AMJ), the peak season of July-September (JAS) and the late season of October-December (OND). During AMJ, EPW (EPC) is associated with a significant increase of the TC genesis number in the southeastern (southwestern) sub-region of the WNP, but no class of El Niño-Southern Oscillation (ENSO) events shows a significant change in the TC lifetime and intensity. During JAS, EPW corresponds to an increase (decrease) of the TC genesis number in the southeastern (northwestern) sub-region, but CPW shows no significant change. EPC increases the TC genesis in the northwestern and northeastern sub-regions and decreases the genesis in the southwestern sub-region, whereas CPC suppresses the genesis in the southeastern sub-region. Both the lifetime and intensity of TCs are increased in EPW, but only a shortened lifetime is seen for CPC. During OND, EPW reduces the TC genesis in the southwestern and northwestern sub-regions, whereas CPW enhances the genesis in the southeastern sub-region. Over the South China Sea, CPW and CPC show a significant decrease and increase of the TC genesis, respectively. The TC lifetime is significantly longer in both EPW and CPW and shorter in EPC, and TCs tend to be more (less) intense in EPW (CPC). All of these variations are consistent with the development of ENSO-related SST anomalies during different seasons and are supported by distributions of the genesis potential index--a combination of large-scale oceanic and atmospheric factors that affect TC activity. TCs in the WNP mainly take the straight westward, northwestward and recurving tracks. During AMJ of EPW years, the TC steering flow patterns favor the recurving track and suppress the straight westward and northwestward tracks. During JAS, EPW is associated with

  12. Seasonal modulations of different impacts of two types of ENSO events on tropical cyclone activity in the western North Pacific

    NASA Astrophysics Data System (ADS)

    Wang, Chunzai; Li, Chunxiang; Mu, Mu; Duan, Wansuo

    2013-06-01

    The paper examines different impacts of eastern Pacific warm/cold (EPW/EPC) and central Pacific warm/cold (CPW/CPC) events on tropical cyclones (TCs) in the western North Pacific (WNP) by considering the early season of April-June (AMJ), the peak season of July-September (JAS) and the late season of October-December (OND). During AMJ, EPW (EPC) is associated with a significant increase of the TC genesis number in the southeastern (southwestern) sub-region of the WNP, but no class of El Niño-Southern Oscillation (ENSO) events shows a significant change in the TC lifetime and intensity. During JAS, EPW corresponds to an increase (decrease) of the TC genesis number in the southeastern (northwestern) sub-region, but CPW shows no significant change. EPC increases the TC genesis in the northwestern and northeastern sub-regions and decreases the genesis in the southwestern sub-region, whereas CPC suppresses the genesis in the southeastern sub-region. Both the lifetime and intensity of TCs are increased in EPW, but only a shortened lifetime is seen for CPC. During OND, EPW reduces the TC genesis in the southwestern and northwestern sub-regions, whereas CPW enhances the genesis in the southeastern sub-region. Over the South China Sea, CPW and CPC show a significant decrease and increase of the TC genesis, respectively. The TC lifetime is significantly longer in both EPW and CPW and shorter in EPC, and TCs tend to be more (less) intense in EPW (CPC). All of these variations are consistent with the development of ENSO-related SST anomalies during different seasons and are supported by distributions of the genesis potential index—a combination of large-scale oceanic and atmospheric factors that affect TC activity. TCs in the WNP mainly take the straight westward, northwestward and recurving tracks. During AMJ of EPW years, the TC steering flow patterns favor the recurving track and suppress the straight westward and northwestward tracks. During JAS, EPW is associated with

  13. 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

  14. 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

  15. Biofilm detection by wound blotting can predict slough development in pressure ulcers: A prospective observational study.

    PubMed

    Nakagami, Gojiro; Schultz, Gregory; Gibson, Daniel J; Phillips, Priscilla; Kitamura, Aya; Minematsu, Takeo; Miyagaki, Tomomitsu; Hayashi, Akitatsu; Sasaki, Sanae; Sugama, Junko; Sanada, Hiromi

    2016-12-26

    Bacteria have been found to form multicellular aggregates which have collectively been termed "biofilms." It is hypothesized that biofilm formation is a means to protect bacterial cells including protection form the immune response of humans. This protective mechanism is believed to explain persistent chronic wound infections. At times, the biofilms are abundant enough to see, and remove by simple wiping. However, recent evidence has shown that the removal of these visible portions are not sufficient, and that biofilms can continue to form even with daily wiping. In this work, we tested an approach to detect the biofilms which are present after clinically wiping or sharp wound debridement. Our method is based on a variation of impression cytology in which a nitrocellulose membrane was used to collect surface biofilm components, which were then differentially stained. In this prospective study, members of an interdisciplinary pressure ulcer team at a university hospital tested our method's ability to predict the generation of wound slough in the week that followed each blotting. A total of 70 blots collected from 23 pressure ulcers produced 27 wounds negative for staining and 43 positive. In the negative blots 55.6% were found to have decreased wound slough, while 81.4% with positive staining had either increase or unchanged wound slough generation. These results lead to an odds ratio of positive blotting cases of 9.37 (95% confidence intervals: 2.47-35.5, p = 0.001) for slough formation; suggesting that the changes in wound slough formation can be predicted clinically using a non-invasive wound blotting method.

  16. 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

  17. Detection of Long Non-coding RNA Expression by Non-radioactive Northern Blots

    PubMed Central

    Hu, Xiaowen; Feng, Yi; Hu, Zhongyi; Zhang, Youyou; Yuan, Chao-Xing; Xu, Xiaowei; Zhang, Lin

    2016-01-01

    With the advances in sequencing technology and transcriptome analysis, it is estimated that up to 75% of the human genome is transcribed into RNAs. This finding prompted intensive investigations on the biological functions of non-coding RNAs and led to very exciting discoveries of microRNAs as important players in disease pathogenesis and therapeutic applications. Research on long non-coding RNAs (lncRNAs) is in its infancy, yet a broad spectrum of biological regulations has been attributed to lncRNAs. As a novel class of RNA transcripts, the expression level and splicing variants of lncRNAs are various. Northern blot analysis can help us learn about the identity, size, and abundance of lncRNAs. Here we describe how to use northern blot to determine lncRNA abundance and identify different splicing variants of a given lncRNA. PMID:26721491

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

    PubMed Central

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

    1991-01-01

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

  19. Direct detection of molluscum contagiosum virus in clinical specimens by dot blot hybridization.

    PubMed Central

    Hurst, J W; Forghani, B; Chan, C S; Oshiro, L; Darai, G

    1991-01-01

    A dot blot hybridization protocol was developed for the direct detection of molluscum contagiosum virus (MCV) DNA in clinical specimens submitted for virus isolation. Samples were concentrated by high-speed centrifugation and treated with proteinase K; this was followed by a single phenol-chloroform extraction step. The DNA was denatured, and the entire volume was spotted onto a nitrocellulose membrane. A biotinylated DNA probe specific for the BamHI-C region of MCV type 1 was used for hybridization. Evidence of MCV DNA was visualized by using streptavidin alkaline phosphatase conjugate and 5-bromo-4-chloro-3-indolyl phosphate-nitroblue tetrazolium as the substrate. Results showed that nonspecific hybridization does not occur with herpes simplex virus- or orf virus-infected clinical specimens and that dot blotting is more sensitive and reproducible than electron microscopy. Images PMID:1774321

  20. 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.

  1. Using genomic slot blot hybridization to assess intergeneric Saccharum x Erianthus hybrids (Andropogoneae - Saccharinae).

    PubMed

    Besse, P; McIntyre, C L; Burner, D M; Almeida, C G

    1997-08-01

    The use of genomic slot blot hybridization enabled the differentiation of hybrids from selfs in Saccharum x Erianthus intergeneric crosses in which Saccharum was used as the female parent. Based on the genomic in situ hybridization technique, slot blots of DNA from the parents and the progeny were blocked with the Saccharum parent DNA and hybridized with the labelled male Erianthus genomic DNA. This technique allowed a rapid screening for hybrids and was sensitive enough to detect a 1/20 dilution of Erianthus in Saccharum DNA, which should enable the detection of most partial hybrids. The genomic slot blot hybridization technique was shown to be potentially useful for assessing crosses involving Saccharum species with either Old World Erianthus section Ripidium or North American Erianthus (= Saccharum) species. The effectiveness of the technique was assessed on 144 progeny of a Saccharum officinarum x Erianthus arundinaceus cross, revealing that 43% of the progeny were selfs. The importance of this test as a tool to support intergeneric breeding programs is discussed.

  2. Western Diet Chow Consumption in Rats Induces Striatal Neuronal Activation While Reducing Dopamine Levels without Affecting Spatial Memory in the Radial Arm Maze.

    PubMed

    Nguyen, Jason C D; Ali, Saher F; Kosari, Sepideh; Woodman, Owen L; Spencer, Sarah J; Killcross, A Simon; Jenkins, Trisha A

    2017-01-01

    Rats fed high fat diets have been shown to be impaired in hippocampal-dependent behavioral tasks, such as spatial recognition in the Y-maze and reference memory in the Morris water maze (MWM). It is clear from previous studies, however, that motivation and reward factor into the memory deficits associated with obesity and high-fat diet consumption, and that the prefrontal cortex and striatum and neurotransmitter dopamine play important roles in cognitive performance. In this series of studies we extend our research to investigate the effect of a high fat diet on striatal neurochemistry and performance in the delayed spatial win-shift radial arm maze task, a paradigm highly reliant on dopamine-rich brain regions, such as the striatum after high fat diet consumption. Memory performance, neuronal activation and brain dopaminergic levels were compared in rats fed a "Western" (21% fat, 0.15% cholesterol) chow diet compared to normal diet (6% fat, 0.15% cholesterol)-fed controls. Twelve weeks of dietary manipulation produced an increase in weight in western diet-fed rats, but did not affect learning and performance in the delayed spatial win-shift radial arm maze task. Concurrently, there was an observed decrease in dopamine levels in the striatum and a reduction of dopamine turnover in the hippocampus in western diet-fed rats. In a separate cohort of rats Fos levels were measured after rats had been placed in a novel arena and allowed to explore freely. In normal rats, this exposure to a unique environment did not affect neuronal activation. In contrast, rats fed a western diet were found to have significantly increased Fos expression in the striatum, but not prefrontal cortex or hippocampus. Our study demonstrates that while western diet consumption in rats produces weight gain and brain neuronal and neurotransmitter changes, it did not affect performance in the delayed spatial win-shift paradigm in the radial arm maze. We conclude that modeling the cognitive decline

  3. 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

  4. Asymmetric response of tropical cyclone activity to global warming over the North Atlantic and western North Pacific from CMIP5 model projections

    NASA Astrophysics Data System (ADS)

    Park, Doo-Sun R.; Ho, Chang-Hoi; Chan, Johnny C. L.; Ha, Kyung-Ja; Kim, Hyeong-Seog; Kim, Jinwon; Kim, Joo-Hong

    2017-01-01

    Recent improvements in the theoretical understanding of the relationship between tropical cyclones (TCs) and their large-scale environments have resulted in significant improvements in the skill for forecasting TC activity at daily and seasonal time-scales. However, future changes in TC activity under a warmer climate remain uncertain, particularly in terms of TC genesis locations and subsequent pathways. Applying a track-pattern-based statistical model to 22 Coupled Model Intercomparison Project Phase 5 (CMIP5) model runs for the historical period and the future period corresponding to the Representative Concentration Pathway 8.5 emissions scenarios, this study shows that in future climate conditions, TC passage frequency will decrease over the North Atlantic, particularly in the Gulf of Mexico, but will increase over the western North Pacific, especially that hits Korea and Japan. Unlike previous studies based on fine-resolution models, an ensemble mean of CMIP5 models projects an increase in TC activity in the western North Pacific, which is owing to enhanced subtropical deep convection and favorable dynamic conditions therein in conjunction with the expansion of the tropics and vice versa for the North Atlantic. Our results suggest that North America will experience less TC landfalls, while northeast Asia will experience more TCs than in the present-day climate.

  5. Asymmetric response of tropical cyclone activity to global warming over the North Atlantic and western North Pacific from CMIP5 model projections.

    PubMed

    Park, Doo-Sun R; Ho, Chang-Hoi; Chan, Johnny C L; Ha, Kyung-Ja; Kim, Hyeong-Seog; Kim, Jinwon; Kim, Joo-Hong

    2017-01-30

    Recent improvements in the theoretical understanding of the relationship between tropical cyclones (TCs) and their large-scale environments have resulted in significant improvements in the skill for forecasting TC activity at daily and seasonal time-scales. However, future changes in TC activity under a warmer climate remain uncertain, particularly in terms of TC genesis locations and subsequent pathways. Applying a track-pattern-based statistical model to 22 Coupled Model Intercomparison Project Phase 5 (CMIP5) model runs for the historical period and the future period corresponding to the Representative Concentration Pathway 8.5 emissions scenarios, this study shows that in future climate conditions, TC passage frequency will decrease over the North Atlantic, particularly in the Gulf of Mexico, but will increase over the western North Pacific, especially that hits Korea and Japan. Unlike previous studies based on fine-resolution models, an ensemble mean of CMIP5 models projects an increase in TC activity in the western North Pacific, which is owing to enhanced subtropical deep convection and favorable dynamic conditions therein in conjunction with the expansion of the tropics and vice versa for the North Atlantic. Our results suggest that North America will experience less TC landfalls, while northeast Asia will experience more TCs than in the present-day climate.

  6. Asymmetric response of tropical cyclone activity to global warming over the North Atlantic and western North Pacific from CMIP5 model projections

    PubMed Central

    Park, Doo-Sun R.; Ho, Chang-Hoi; Chan, Johnny C. L.; Ha, Kyung-Ja; Kim, Hyeong-Seog; Kim, Jinwon; Kim, Joo-Hong

    2017-01-01

    Recent improvements in the theoretical understanding of the relationship between tropical cyclones (TCs) and their large-scale environments have resulted in significant improvements in the skill for forecasting TC activity at daily and seasonal time-scales. However, future changes in TC activity under a warmer climate remain uncertain, particularly in terms of TC genesis locations and subsequent pathways. Applying a track-pattern-based statistical model to 22 Coupled Model Intercomparison Project Phase 5 (CMIP5) model runs for the historical period and the future period corresponding to the Representative Concentration Pathway 8.5 emissions scenarios, this study shows that in future climate conditions, TC passage frequency will decrease over the North Atlantic, particularly in the Gulf of Mexico, but will increase over the western North Pacific, especially that hits Korea and Japan. Unlike previous studies based on fine-resolution models, an ensemble mean of CMIP5 models projects an increase in TC activity in the western North Pacific, which is owing to enhanced subtropical deep convection and favorable dynamic conditions therein in conjunction with the expansion of the tropics and vice versa for the North Atlantic. Our results suggest that North America will experience less TC landfalls, while northeast Asia will experience more TCs than in the present-day climate. PMID:28134343

  7. Active geodynamics of the central Mediterranean Sea: Tensional tectonic evidences in western Sicily from mantle-derived helium

    NASA Astrophysics Data System (ADS)

    Caracausi, A.; Favara, R.; Italiano, F.; Nuccio, P. M.; Paonita, A.; Rizzo, A.

    2005-02-01

    We report results on the measured high 3He/4He isotope ratio in western Sicily, interpreted together with the heat data. The study of this sector of the Europe-Africa interaction is crucial to a better understanding of the tectonics and the geodynamical evolution of the central Mediterranean area. The estimated mantle-derived helium fluxes in the investigated areas are up to 2-3 orders of magnitude greater than those of a stable continental area. The highest flux, found in the southernmost area near the Sicily Channel, where recent eruptions of the Ferdinandea Island occurred 20 miles out to sea off Sciacca, has been associated with a clear excess of heat flow. Our results indicate that there is an accumulation of magma below the continental crust of western Sicily that is possibly intruding and out-gassing through roughly N-S trending deep fault systems linked to the mantle, that have an extensional component. Although the identification of these faults is not sufficiently constrained by our data, they could possibly be linked to the pre-existing faults that originated during the Mesozoic extensional-transtensional tectonic phases.

  8. 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

  9. An improved method for Southern DNA and Northern RNA blotting using a Mupid-2 Mini-Gel electrophoresis unit.

    PubMed

    Furuya, Hirokazu; Yamada, Takeshi; Ikezoe, Koji; Ohyagi, Yasumasa; Fukumaki, Yasuyuki; Fujii, Naoki

    2006-08-31

    An improved method for Southern DNA and Northern RNA blotting using the Mupid-2 Mini-Gel System is described. We get sharp and clear bands in Southern and Northern blotting after only 30 min short gel electrophoresis instead of the several hours large gel electrophoresis of conventional methods. The high electrical voltage with a pulse-like current of the Mupid-2 Mini-Gel System also allows reduction of the amount of formaldehyde, a harmful reagent, from the gel running buffer in RNA blotting. This minor modification of DNA and RNA blotting technique enables us to perform the complete experimental procedure more quickly economically in less space, than conventional Southern and Northern blotting, as well as using an extremely small amount of formaldehyde in RNA blotting.

  10. 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.

  11. Activity of spores and extracellular proteins from six Cry+ strains and a Cry- strain of Bacillus thuringiensis subsp. kurstaki against the western spruce budworm, Choristoneura occidentalis (Lepidoptera: Tortricidae).

    PubMed

    Kalmykova, Galina; Burtseva, Ljudmila; Milne, Ross; van Frankenhuyzen, Kees

    2009-05-01

    We characterized insecticidal activity of previously untested strains of Bacillus thuringiensis kurstaki belonging to two crystal serovars (K-1 and K-73) against the western spruce budworm (Choristoneura occidentalis Freeman 1967). By testing various components, we demonstrated that spores play a critical role in the pathogenesis of each strain. Spore-free crystals caused low mortality and purified spores were generally not toxic. The addition of spores to purified protoxin increased toxicity several hundred-fold, regardless of the parental strain from which the spores or protoxins were derived. The crystal and spore components did not account for full insecticidal activity of whole sporulated cultures owing to the toxicity of soluble proteins that are secreted during cell growth. We observed a marked difference in toxicity of secreted proteins between the K-1 and K-73 type strains, with the K-1 preparations causing much higher mortality, mass reduction, and inhibition of pupation. There was a consistent correlation between relative toxicity of secreted protein preparations and the presence and quantity of the Vip3A protein, suggesting that this protein contributes to the virulence of B. thuringiensis subsp. kurstaki in western spruce budworm larvae. However, other virulence factors have to be invoked to explain the synergizing effect of spores from both K-1 and K-73 strains on Cry protein toxicity.

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

    SciTech Connect

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

    1994-09-01

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

  13. 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.

  14. Selection of DNA aptamers against VEGF165 using a protein competitor and the aptamer blotting method.

    PubMed

    Hasegawa, Hijiri; Sode, Koji; Ikebukuro, Kazunori

    2008-05-01

    Two DNA aptamers against a tumor marker protein, human vascular endothelial growth factor (VEGF(165)) were identified. In the screening process, another protein was used as the competitor to isolate those aptamers that have high specificity for the target. In addition, we evaluated the affinities of the enriched library by means of aptamer blotting. The isolated aptamers bound to VEGF(165) with a K(d) value in the range of a few hundred nanomoles, and did not bind to the competitor. This selection method enabled us to efficiently select the specific aptamers against the target protein. These specific aptamers would be useful sensor elements for cancer diagnosis.

  15. Western Diet Chow Consumption in Rats Induces Striatal Neuronal Activation While Reducing Dopamine Levels without Affecting Spatial Memory in the Radial Arm Maze

    PubMed Central

    Nguyen, Jason C. D.; Ali, Saher F.; Kosari, Sepideh; Woodman, Owen L.; Spencer, Sarah J.; Killcross, A. Simon; Jenkins, Trisha A.

    2017-01-01

    Rats fed high fat diets have been shown to be impaired in hippocampal-dependent behavioral tasks, such as spatial recognition in the Y-maze and reference memory in the Morris water maze (MWM). It is clear from previous studies, however, that motivation and reward factor into the memory deficits associated with obesity and high-fat diet consumption, and that the prefrontal cortex and striatum and neurotransmitter dopamine play important roles in cognitive performance. In this series of studies we extend our research to investigate the effect of a high fat diet on striatal neurochemistry and performance in the delayed spatial win-shift radial arm maze task, a paradigm highly reliant on dopamine-rich brain regions, such as the striatum after high fat diet consumption. Memory performance, neuronal activation and brain dopaminergic levels were compared in rats fed a “Western” (21% fat, 0.15% cholesterol) chow diet compared to normal diet (6% fat, 0.15% cholesterol)-fed controls. Twelve weeks of dietary manipulation produced an increase in weight in western diet-fed rats, but did not affect learning and performance in the delayed spatial win-shift radial arm maze task. Concurrently, there was an observed decrease in dopamine levels in the striatum and a reduction of dopamine turnover in the hippocampus in western diet-fed rats. In a separate cohort of rats Fos levels were measured after rats had been placed in a novel arena and allowed to explore freely. In normal rats, this exposure to a unique environment did not affect neuronal activation. In contrast, rats fed a western diet were found to have significantly increased Fos expression in the striatum, but not prefrontal cortex or hippocampus. Our study demonstrates that while western diet consumption in rats produces weight gain and brain neuronal and neurotransmitter changes, it did not affect performance in the delayed spatial win-shift paradigm in the radial arm maze. We conclude that modeling the cognitive

  16. 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

  17. Kinase inhibitory, haemolytic and cytotoxic activity of three deep-water sponges from North Western Australia and their fatty acid composition.

    PubMed

    Zivanovic, Ana; Pastro, Natalie J; Fromont, Jane; Thomson, Murray; Skropeta, Danielle

    2011-12-01

    The c-AMP dependent protein kinase (PKA) inhibition, haemolytic activity, and cytoxicity of 21 extracts obtained from North Western Australian sponges collected from depths of 84-135 m were investigated. Hexane extracts from Ircinia/Sarcotragus sp. and Geodia sp. displayed PKA inhibitory activities of 100 and 97% respectively (at 100 microg/mL), while aq. methanol extracts from Haliclona sp. exhibited potent haemolytic activity (75%) and hexane extracts from Geodia sp. were highly toxic (88%) to the brine shrimp Artemia franciscana. As the non-polar extracts gave the greatest PKA inhibition, these were further analysed by GC-MS and 29 fatty acids were identified in the highest proportions in Ircinia/Sarcotragus sp. > Haliclona sp. > Geodia sp. In contrast to shallow-water sponges that are dominated by polyunsaturated fatty acids with a high percentage of long chain fatty acids, LCFAs (C24-C30), the deep-sea sponges investigated herein were all found to be rich in saturated fatty acids, in particular C14-C20 fatty acids, including odd and branched chain fatty acids, with only low levels (0-10%) of LCFAs. Screening of the PKA inhibitory activity of a series of commercially available fatty acids identified C14-C18 fatty acids as possessing significant PKA inhibitory activity that may contribute to the activity observed in the sponges studied.

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

    NASA Astrophysics Data System (ADS)

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

    2010-01-01

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

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

    SciTech Connect

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

    1982-06-01

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

  20. A simple DNA recombination screening method by RT-PCR as an alternative to Southern blot.

    PubMed

    Albers, Eliene; Sbroggiò, Mauro; Martin-Gonzalez, Javier; Avram, Alexandra; Munk, Stephanie; Lopez-Contreras, Andres J

    2017-01-19

    The generation of genetically engineered mouse models (GEMMs), including knock-out (KO) and knock-in (KI) models, often requires genomic screening of many mouse ES cell (mESC) clones by Southern blot. The use of large targeting constructs facilitates the recombination of exogenous DNA in a specific genomic locus, but limits the detection of its correct genomic integration by standard PCR methods. Genomic Long Range PCR (LR-PCR), using primers adjacent to the homology arms, has been used as an alternative to radioactive-based Southern blot screenings. However, LR-PCRs are often difficult and render many false positive and false negative results. Here, we propose an alternative screening method based on the detection of a genetic modification at the mRNA level, which we successfully optimized in two mouse models. This screening method consists of a reverse-transcription PCR (RT-PCR) using primers that match exons flanking the targeting construct. The detection of the expected modification in this PCR product confirms the integration at the correct genomic location and shows that the mutant mRNA is expressed. This is a simple and sensitive strategy to screen locus-specific recombination of targeting constructs which can also be useful to screen KO and KI mutant mice or cell lines including those generated by CRISPR/Cas9.

  1. Capillary blotting of glycosaminoglycans on nitrocellulose membranes after agarose-gel electrophoresis separation.

    PubMed

    Volpi, Nicola; Maccari, Francesca

    2009-01-01

    A method for the blotting and immobilizing of several nonsulfated and sulfated complex polysaccharides on membranes made hydrophilic and positively charged by cationic detergent after their separation by conventional agarose gel electrophoresis is illustrated. This new approach to the study of glycosaminoglycans (GAGs) utilizes the capacity of agarose gel electrophoresis to separate single species of polysaccharides from mixtures and the membrane technology for further preparative and analytical uses.Nitrocellulose membranes are derivatized with the cationic detergent cetylpyridinium chloride and mixtures of GAGs are capillary blotted after their separation in agarose gel electrophoresis. Single purified species of variously sulfated polysaccharides are transferred on derivatized membranes with an efficiency of 100% and stained with alcian blue (irreversible staining) and toluidine blue (reversible staining). This enables a lower amount limit of detection of 0.1 microg. Nonsulfated polyanions, for example hyaluronic acid, may also be transferred to membranes with a limit of detection of approximately 0.1-0.5 microg after irreversible or reversible staining. The membranes may be stained with reversible staining and the same lanes are used for immunological detection or other applications.

  2. 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.

  3. 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.

  4. Using ground-penetrating radar, topography and classification of vegetation to model the sediment and active layer thickness in a periglacial lake catchment, western Greenland

    NASA Astrophysics Data System (ADS)

    Petrone, Johannes; Sohlenius, Gustav; Johansson, Emma; Lindborg, Tobias; Näslund, Jens-Ove; Strömgren, Mårten; Brydsten, Lars

    2016-11-01

    The geometries of a catchment constitute the basis for distributed physically based numerical modeling of different geoscientific disciplines. In this paper results from ground-penetrating radar (GPR) measurements, in terms of a 3-D model of total sediment thickness and active layer thickness in a periglacial catchment in western Greenland, are presented. Using the topography, the thickness and distribution of sediments are calculated. Vegetation classification and GPR measurements are used to scale active layer thickness from local measurements to catchment-scale models. Annual maximum active layer thickness varies from 0.3 m in wetlands to 2.0 m in barren areas and areas of exposed bedrock. Maximum sediment thickness is estimated to be 12.3 m in the major valleys of the catchment. A method to correlate surface vegetation with active layer thickness is also presented. By using relatively simple methods, such as probing and vegetation classification, it is possible to upscale local point measurements to catchment-scale models, in areas where the upper subsurface is relatively homogeneous. The resulting spatial model of active layer thickness can be used in combination with the sediment model as a geometrical input to further studies of subsurface mass transport and hydrological flow paths in the periglacial catchment through numerical modeling. The data set is available for all users via the PANGAEA database, doi:10.1594/PANGAEA.845258.

  5. [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.

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

    PubMed

    Bucała, Anna

    2014-06-01

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

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

    PubMed

    Kawahara, Nobuo

    2011-03-01

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

  8. Human T-cell lymphotropic virus type I (HTLV-I) confirmed by PCR-Southern blot and sequencing analysis in a woman with tropical spastic paraparesis.

    PubMed

    Perandin, F; Cariani, A; Bonfanti, C; Trainini, L; Magoni, M; Manca, N

    2006-09-01

    Human T-cell lymphotropic virus type I (HTLV-I) is a human retrovirus and the aetiological agent of a progressive neurological disease called tropical spastic paraparesis/HTLV-I-associated myelopathy (TSP/HAM), as confirmed by evidence accumulated in HTLV-I seroprevalence studies. TSP/HAM is rarely diagnosed in Italy, given the low prevalence of HTLV-I in the population. TSP/HAM begins insidiously in the fourth decade, mainly with spastic paraparesis of the lower extremities and positive Babinski reflex, as well as interfering with bowel and bladder functions. In this study we report the clinical, virological and haemato chemical data of a 54-year-old woman, born in the Ivory Cost, with symptoms suggestive of TSP. The presence of HTLV-I infection was demonstrated by the detection of antibodies in serum and in cerebrospinal fluid by immunoenzymatic assay and Western blot analysis. In addition, viral isolation was carried out in peripheral blood cells, and the presence of HTLV-I proviral DNA was confirmed by polymerase chain reaction/Southern blot and sequencing analysis. According to our results, HTLV-I testing might be useful when TSP/HAM is suspected.

  9. Quantitation of proteoglycans as glycosaminoglycans in biological fluids using an alcian blue dot blot analysis.

    PubMed

    Björnsson, S

    1998-02-15

    A method for quantitation of intact proteoglycans as GAGs in biological fluids (blood plasma, synovial fluid) or 4 M guanidine extracts of tissues has been published previously (S. Björnsson, Anal. Biochem. 210, 282-291, 1993). The method is based on the specific interaction between sulfated polymers and the tetravalent cationic dye Alcian blue at pH 1.5 in 0.4 M guanidine-HCl and in the presence of 0.25% Triton. The absorbance assay has a measuring range of 1-20 microgram of glycosaminoglycan (GAG) which is not sensitive enough to measure the low contents of proteoglycans in blood plasma, urine, or wound fluid. A dot blot assay is now described in which the Alcian blue-GAG complexes are collected on a polyvinylidene fluoride membrane, by filtration in a dot blot apparatus, and the stain is quantitated as reflectance by scanning and densitometry. The assay requires 10 microliter of sample and has a measuring range of 10-800 ng of GAG, corresponding to a concentration of 1-80 mg/liter, suitable for proteoglycans in biological fluids. The procedures for chemistry, scanning, densitometry, and curve fitting were each evaluated separately. The error contributed by chemistry accounted for a minor portion of the imprecision. The imprecision contributed by scanning was the most important source of within-run and between-run imprecision, and was caused by inequalities of the charge-coupled device along the scanning arm. Unexpectedly, curve fitting was also a major source of total imprecision in dot blot quantitation and differed with the type of equation used. The between-run imprecision calculated as CV (SD/mean . 100) was 13.0% at 8 mg/liter. The response of the assay was identical for six different commercial preparations of GAGs (chondroitin-4-sulfate, chondroitin-6-sulfate, dermatan sulfate, keratan sulfate, heparan sulfate, and heparin) despite differences in degree of sulfation known to exist. There was no positive or negative interference by blood plasma, apart

  10. Application of a Dot Blot Hybridization Platform to Assess Streptococcus uberis Population Structure in Dairy Herds

    PubMed Central

    Albuquerque, Pedro; Ribeiro, Niza; Almeida, Alexandre; Panschin, Irena; Porfirio, Afonso; Vales, Marta; Diniz, Francisca; Madeira, Helena; Tavares, Fernando

    2017-01-01

    Streptococcus uberis is considered one of the most important pathogens associated with bovine mastitis. While traditionally acknowledged as an environmental pathogen, S. uberis has been shown to adopt a contagious epidemiological pattern in several dairy herds. Since different control strategies are employed depending on the mode of transmission, in-depth studies of S. uberis populations are essential to determine the best practices to control this pathogen. In this work, we optimized and validated a dot blot platform, combined with automatic image analysis, to rapidly assess the population structure of infective S. uberis, and evaluated its efficiency when compared to multilocus sequence analysis (MLSA) genotyping. Two dairy herds with prevalent S. uberis infections were followed in a 6 month period, in order to collect and characterize isolates from cows with persistent infections. These herds, located in Portugal (Barcelos and Maia regions), had similar management practices, with the herd from Barcelos being smaller and having a better milking parlor management, since infected cow segregation was immediate. A total of 54 S. uberis isolates were obtained from 24 different cows from the two herds. To overcome operator-dependent analysis of the dot blots and increase the technique's consistency and reliability, the hybridization signals were converted into probability values, with average probabilities higher than 0.5 being considered positive results. These data allowed to confirm the isolates' identity as S. uberis using taxa-specific markers and to determine the presence of virulence- and antibiotic resistance-related genes. In addition, MLSA allowed to disclose the most prevalent S. uberis clonal lineages in both herds. Seven different clusters were identified, with Barcelos showing a high clonal diversity and Maia a dominant lineage infecting most cows, suggesting distinct epidemiological patterns, with S. uberis displaying an environmental or contagious

  11. 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.

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  13. Reverse line blot hybridization used to identify hemoprotozoa in Minorcan cattle.

    PubMed

    Almeria, Sonia; Castella, Joaquim; Ferrer, David; Gutierrez, Juan Francisco; Estrada-Pena, Agustin; Sparagano, Olivier

    2002-10-01

    Piroplasmosis, a tick-borne protozoal disease, is an important disease affecting domestic and wild animals. We performed PCR-based reverse line blot hybridization (RLB) assays on blood samples obtained from 133 cattle exposed to ticks in field conditions in Minorca (Balearic Islands, Spain) in three different seasons. The oligonucleotides used were those for Theileria annulata, T. buffeli, T. taurotragi, T. velifera, Babesia bigemina, B. bovis, B. divergens, and B. major. The RLB technique allowed the simultaneous identification of T. annulata, T. buffeli, B. bigemina, and B. bovis as the piroplasms present in cattle in Minorca. Of the 133 animals, only 4 were not infected by any of the studied parasites. The results indicated endemic piroplasm infection in cattle in Minorca; especially important was the presence of T. annulata. The RLB was highly sensitive and allowed the simultaneous detection and identification of the Theileria and Babesia species in carrier cattle, which cannot be achieved by classical identification methods.

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

    PubMed

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

    2015-09-15

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

  15. Comparative study on the antioxidant activities of extracts of Coreopsis tinctoria flowering tops from Kunlun Mountains, Xinjiang, north-western China.

    PubMed

    Yao, Xincheng; Gu, Chengzhi; Tian, Liping; Wang, Xinbing; Tang, Hui

    2016-01-01

    Coreopsis tinctoria flowering tops (CTFs) from the Kunlun Mountains in Xinjing (north-western China) have been used for tea production for about a century. This study aims to assess the antioxidant activities and total phenolic, flavonoid and proanthocyanidin contents of various solvent extracts of CTF. CTF was extracted using n-hexane, chloroform, ethyl acetate, n-butanol, 75% aqueous ethanol (AEE) and water. The antioxidant activities of the CTF extracts were investigated through DPPH, ABTS, *OH, *O2(-), total antioxidant capacity and reducing power assays. The results showed that n-butanol extract showed the highest contents of total phenols and flavonoids, with DPPH, ABTS and *OH radical-scavenging activities with IC50 values of 134, 90.72 and 13.8 μg mL(-1), respectively. The AEE demonstrated the strongest DPPH and ABTS radical-scavenging activities, with IC50 values of 103 and 75.16 μg mL(-1), respectively. Given its high antioxidant effect, CTF is a good source of natural antioxidants or functional food materials.

  16. 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

  17. Optimization of dot blot method to detect bcr/abl transcripts in chronic myeloid leukemia

    SciTech Connect

    Tharapel, S.A.; Zhao, J.

    1994-09-01

    Detection of abl-bcr fusion transcripts using molecular methodologies is becoming an attractive alternative (or supplement) to traditional cytogenetics in identifying the Philadelphia (Ph) chromosome. Among these methods, RT-PCR technique has provided an extremely powerful tool for improving the detection of bcr/abl translocations through enzymatic amplification of the reverse-transcribed cDNA. The analysis of PCR products can be accomplished by a number of techniques including dot blot following liquid-phase hybridization. In order to render the detection of PCR products more simple, accurate and efficient, and therefore more amenable for the clinical laboratory routine use, we optimized several parameters of the procedure. (1) We discovered that with the starting material of 1 ug of total RNA, the amount of the final PCR amplified products was linear to the PCR cycles between 20 to 30 cycles. Since the dot blot procedure does not separate the amplified products according to their sizes, increased background would increase the false positive rate. (2) If a detection sensitivity of 1 in 10{sup 3} cells is sufficient, then the nested or a second PCR amplification is not necessary. (3) Starting material more than 5 ug of total RNA would decrease the amplification efficiency and therefore compromise the sensitivity. (4) Ten minutes of hybridization gave equal signal intensity as 24 hours. (5) The ionic strength and temperature in the washing step were also tested. Upon optimization of each parameter, the detection procedure was tested on 18 clinical samples. Compared to the procedures that are currently available, our optimized procedure is less time consuming, has higher sensitivity and lower false positive rate. This method has the potential to be automated and therefore can be used as a screening method for Ph chromosome in high volume settings.

  18. Rapid detection and differentiation of important Campylobacter spp. in poultry samples by dot blot and PCR.

    PubMed

    Fontanot, Marco; Iacumin, Lucilla; Cecchini, Francesca; Comi, Giuseppe; Manzano, Marisa

    2014-10-01

    The detection of Campylobacter, the most commonly reported cause of foodborne gastroenteritis in the European Union, is very important for human health. The most commonly recognised risk factor for infection is the handling and/or consumption of undercooked poultry meat. The methods typically applied to evaluate the presence/absence of Campylobacter in food samples are direct plating and/or enrichment culture based on the Horizontal Method for Detection and Enumeration of Campylobacter spp. (ISO 10272-1B: 2006) and PCR. Molecular methods also allow for the detection of cells that are viable but cannot be cultivated on agar media and that decrease the time required for species identification. The current study proposes the use of two molecular methods for species identification: dot blot and PCR. The dot blot method had a sensitivity of 25 ng for detection of DNA extracted from a pure culture using a digoxigenin-labelled probe for hybridisation; the target DNA was extracted from the enrichment broth at 24 h. PCR was performed using a pair of sensitive and specific primers for the detection of Campylobacter jejuni and Campylobacter coli after 24 h of enrichment in Preston broth. The initial samples were contaminated by 5 × 10 C. jejuni cells/g and 1.5 × 10(2)C. coli cells/g, thus the number of cells present in the enrichment broth at 0 h was 1 or 3 cell/g, respectively.

  19. In-situ Stresses, Pore-fluid Pressures and Uplift Erosion in Relation to Active Thrust Faulting in western Taiwan

    NASA Astrophysics Data System (ADS)

    Hung, J.; Yen, P.; Wang, L.

    2012-12-01

    We have studied the in-situ stresses, pore-fluid pressures and amounts of uplift erosion (UE) from petroleum wells drilled in the Hsinchu-Taichung area of western Taiwan Fold-thrust Belt. The average gradient of regional vertical stress (Sv) calculated from formation density logs is about 23 MPa/km. The magnitude of pore pressure (Pp) is estimated from mud pressure, gas cut and repeat formation test (RFT) in reservoir sandstone, and sonic logs. P-wave travel time in shale (STT) is used to determine the fluid-retention depth (ZFRD) which defines current fully compacted sediments with hydrostatic pressures above and undercompacted, overpressured zones below. Regional ZFRD is ~ 3 km except in the Chuhuangkeng anticline, where ZFRD is at shallower depth (~ 2.2 km) and extremely high pore pressure (λ=0.8) is also observed.. Calculated amounts of UE increase from 0.6 to 4.6 km eastward from outer to inner Foothills belt and correspond to stratigraphy downward and depth upward migration of the ZFRD. Along-strike variation of UE is insignificant. Hydraulic fracturing data including leak-off tests (LOTs) and mini-fracs, as well as qualitative data such as mud loss, are used to constrain the minimum horizontal stress (Shmin). The linear gradient of Shmin is about 17~19 MPa/km, relatively less than that of Sv (~23.60 MPa/km). This implies the in-situ stresses are at strike-slip (SHmax>SV>Shmin) to reverse fault considering focal mechanisms of seismicity are dominant by these two stress regimes in the study area. An upper-bound value of the maximum horizontal stress (SHmax) constrained by frictional limits and the coefficient of friction (μ=0.6) can be estimated from Anderson (1951) faulting criterion. Caliper logs from 8 wells are used to calculate the orientations of the maximum horizontal stresses following the definitions of borehole breakout in World Stress Map. The maximum horizontal stress axis is oriented in NW-SE but local variations occur when passing through

  20. 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

  1. Serologic Evidence for Novel Poxvirus in Endangered Red Colobus Monkeys, Western Uganda

    PubMed Central

    Chapman, Colin A.; Cameron, Kenneth; Saj, Tania; Karesh, William B.; Wolfe, Nathan D.; Wong, Scott W.; Dubois, Melissa E.; Slifka, Mark K.

    2008-01-01

    Enzyme-linked immunosorbent assay, Western blot, and virus neutralization assays indicated that red colobus monkeys in Kibale National Park, western Uganda, had antibodies to a virus that was similar, but not identical, to known orthopoxviruses. The presence of a novel poxvirus in this endangered primate raises public health and conservation concerns. PMID:18439366

  2. Chemical variability, antioxidant and antifungal activities of essential oils and hydrosol extract of Calendula arvensis L. from western Algeria.

    PubMed

    Belabbes, Rania; Dib, Mohammed El Amine; Djabou, Nassim; Ilias, Faiza; Tabti, Boufeldja; Costa, Jean; Muselli, Alain

    2017-01-21

    The chemical composition of the essential oils and hydrosol extract from aerial parts of Calendula arvensis L. was investigated using GC-FID and GC/MS. Intra-species variations of the chemical compositions of essential oils from 18 Algerian sample locations were investigated using statistical analysis. Chemical analysis allowed the identification of fifty-three compounds amounting to 92.3-98.5% with yields varied of 0.09-0.36% and the main compounds were zingiberenol 1 (8.7-29.8%), eremoligenol (4.2-12.5%), β-curcumene (2.1-12.5%), zingiberenol 2 (4.6-19.8%) and (E,Z)-farnesol (3.5-23.4%). The study of the chemical variability of essential oils allowed the discrimination of two main clusters confirming that there is a relation between the essential oil compositions and the harvest locations. Different concentrations of essential oil and hydrosol extract were prepared and their antioxidant activity were assessed using three methods (DPPH, FRAP and β-carotene). The results showed that hydrosol extract presented an interesting antioxidant activity. The in vitro antifungal activity of hydrosol extract produced the best antifungal inhibition against Penicillium expansum and Aspergillus niger, while, essential oil was inhibitory at relatively higher concentrations. Results showed that the treatments of pear fruits with essential oil and hydrosol extract presented a very interesting protective activity on disease severity of pears caused by P. expansum. This article is protected by copyright. All rights reserved.

  3. Evidence of sub-vent biosphere: enzymatic activities in 308 °C deep-sea hydrothermal systems at Suiyo seamount, Izu Bonin Arc, Western Pacific Ocean

    NASA Astrophysics Data System (ADS)

    Takano, Yoshinori; Edazawa, Yae; Kobayashi, Kensei; Urabe, Tetsuro; Marumo, Katsumi

    2005-01-01

    A high-temperature deep-sea hydrothermal system related to dacitic arc-volcanism was drilled using a tethered, submarine rock-drill system as a part of the Archaean Park Project. The benthic multi-coring system (BMS) employed allowed for direct sampling of microorganisms, rocks and fluids beneath hydrothermal vents. The samples examined in this study were from sites APSK 05 and APSK 07 on the Suiyo Seamount of the Izu-Bonin Arc in the Pacific Ocean. Based on the vertical distribution of samples derived from this vigorous sub-vent environment, a model of deep-sea subterranean chemistry and biology was determined detailing optimal microbial activities. Deep-sea hydrothermal sub-vent core samples of dacitic arc-volcanism obtained at the Suiyo Seamount, Izu-Bonin Arc, Western Pacific Ocean were analyzed for acid and alkaline phosphatase enzymatic activities. Useful biomarkers of acid phosphatase (ACP) and alkaline phosphatase (ALP) enzymatic activities were positively correlated against each other and was greatest at the partial middle core sequences; ACP and ALP activities determined were as high as 5.10 and 6.80 nmol/min/g rock, respectively. Biochemical indicators of ACP and ALP were consistent with the origin of biogenic amino acids occupied in the sub-vent region and microbial cell number in the fluid. The significant enzymatic activities demonstrated in this study provides crucial evidence that sub-vent regions represent part of the previously unknown extreme-environment biosphere, extending the known subterranean habitable spaces of, for example, extremophilic microbes. This boring trial was first example of discharging high temperature hydrothermal activities at the frontal arc volcanoes.

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

    PubMed

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

    2010-10-15

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

  5. Methane-cycling communities in a permafrost-affected soil on Herschel Island, Western Canadian Arctic: active layer profiling of mcrA and pmoA genes.

    PubMed

    Barbier, Béatrice A; Dziduch, Isabel; Liebner, Susanne; Ganzert, Lars; Lantuit, Hugues; Pollard, Wayne; Wagner, Dirk

    2012-11-01

    In Arctic wet tundra, microbial controls on organic matter decomposition are likely to be altered as a result of climatic disruption. Here, we present a study on the activity, diversity and vertical distribution of methane-cycling microbial communities in the active layer of wet polygonal tundra on Herschel Island. We recorded potential methane production rates from 5 to 40 nmol h(-1) g(-1) wet soil at 10 °C and significantly higher methane oxidation rates reaching values of 6-10 μmol h(-1) g(-1) wet soil. Terminal restriction fragment length polymorphism (T-RFLP) and cloning analyses of mcrA and pmoA genes demonstrated that both communities were stratified along the active layer vertical profile. Similar to other wet Arctic tundra, the methanogenic community hosted hydrogenotrophic (Methanobacterium) as well as acetoclastic (Methanosarcina and Methanosaeta) members. A pronounced shift toward a dominance of acetoclastic methanogens was observed in deeper soil layers. In contrast to related circum-Arctic studies, the methane-oxidizing (methanotrophic) community on Herschel Island was dominated by members of the type II group (Methylocystis, Methylosinus, and a cluster related to Methylocapsa). The present study represents the first on methane-cycling communities in the Canadian Western Arctic, thus advancing our understanding of these communities in a changing Arctic.

  6. 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

  7. A maternal Western diet during gestation and lactation modifies offspring's microbiota activity, blood lipid levels, cognitive responses, and hippocampal neurogenesis in Yucatan pigs.

    PubMed

    Val-Laillet, David; Besson, Marie; Guérin, Sylvie; Coquery, Nicolas; Randuineau, Gwénaëlle; Kanzari, Ameni; Quesnel, Hélène; Bonhomme, Nathalie; Bolhuis, J Elizabeth; Kemp, Bas; Blat, Sophie; Le Huërou-Luron, Isabelle; Clouard, Caroline

    2017-02-06

    A suboptimal early nutritional environment (i.e., excess of energy, sugar, and fat intake) can increase susceptibility to diseases and neurocognitive disorders. The purpose of this study was to investigate in nonobese Yucatan minipigs (Sus scrofa) the impact of maternal diet [standard (SD) vs. Western (WD) diet] during gestation and 25 d of lactation on milk composition, blood metabolism, and microbiota activity of sows (n = 17) and their piglets (n = 65), and on spatial cognition (n = 51), hippocampal plasticity (n = 17), and food preferences/motivation (n = 51) in the progeny. Milk dry matter and lipid content, as well as plasma total cholesterol and free fatty acid (FFA) concentrations (P < 0.05) were higher in WD than in SD sows. Microbiota activity decreased in both WD sows and 100-d-old piglets (P < 0.05 or P < 0.10, depending on short-chain FAs [SCFAs]). At weaning [postnatal day (PND) 25], WD piglets had increased blood triglyceride and FFA levels (P < 0.01). Both SD and WD piglets consumed more of a known SD than an unknown high-fat/-sucrose (HFS) diet (P < 0.0001), but were quicker to obtain HFS rewards compared with SD rewards (P < 0.01). WD piglets had higher working memory (P = 0.015) and reference memory (P < 0.001) scores, which may reflect better cognitive abilities in the task context and a higher motivation for the food rewards. WD piglets had a smaller hippocampal granular cell layer (P = 0.03) and decreased neurogenesis (P < 0.005), but increased cell proliferation (P < 0.001). A maternal WD during gestation and lactation, even in the absence of obesity, has significant consequences for piglets' blood lipid levels, microbiota activity, gut-brain axis, and neurocognitive abilities after weaning.-Val-Laillet, D., Besson, M., Guérin, S., Coquery, N., Randuineau, G., Kanzari, A., Quesnel, H., Bonhomme, N., Bolhuis, J. E., Kemp, B., Blat, S., Le Huërou-Luron, I., Clouard, C. A maternal Western diet during gestation and lactation modifies offspring

  8. Double-blotting: a solution to the problem of nonspecific binding of secondary antibodies in immunoblotting procedures.

    PubMed

    Lasne, Françoise

    2009-01-01

    Nonspecific interactions between blotted proteins and unrelated secondary antibodies generate false positives in immunoblotting techniques. Some procedures have been developed to reduce this adsorption but they may work in specific applications and be ineffective in other ones. "Double-blotting" has been developed to overcome this problem. It consists of interpolating a second blotting step between the usual probings of the blot membrane with the primary antibody and the secondary antibodies. This step, by isolating the primary antibody from the interfering proteins, guarantees the specificity of the probing with the secondary antibody. This method has been developed for the study of erythropoietin in concentrated urine since a strong nonspecific binding of biotinylated secondary antibodies to some urinary proteins is observed using classical immunoblotting protocols. However, its concept makes it usable in other applications that come up against this kind of problem. This method is expected to be especially useful for investigating proteins that are present in minute amounts in complex biological media.

  9. Double-blotting: a solution to the problem of nonspecific binding of secondary antibodies in immunoblotting procedures.

    PubMed

    Lasne, Françoise

    2015-01-01

    Nonspecific interactions between blotted proteins and unrelated secondary antibodies generate false positives in immunoblotting techniques. Some procedures have been developed to reduce this adsorption, but they may work in specific applications and be ineffective in others. "Double-blotting" has been developed to overcome this problem. It consists of interpolating a second blotting step between the usual probings of the blot membrane with the primary antibody and the secondary antibodies. This step, by isolating the primary antibody from the interfering proteins, guarantees the specificity of the probing with the secondary antibody. This method has been developed for the study of erythropoietin in concentrated urine since a strong nonspecific binding of biotinylated secondary antibodies to some urinary proteins is observed using classical immunoblotting protocols. However, its concept makes it usable in other applications that come up against this kind of problem. This method is expected to be especially useful for investigating proteins that are present in minute amounts in complex biological media.

  10. Active shortening along the northern edge of the western Transverse Ranges recorded by deformed fluvial terraces along Santa Cruz Creek in the eastern Santa Ynez Valley, California

    NASA Astrophysics Data System (ADS)

    Tyler, E.

    2011-12-01

    Vertical-axis rotation of the western Transverse Ranges in southern California was accompanied by folding and reverse faulting at the northern edge of the rotated domain during late Miocene to Pliocene time. Deformed Pleistocene strata in the Santa Ynez Valley suggest that this rotational folding and faulting is still occurring. Fluvial terraces are present along several major drainages that cross the faults and folds and provide an opportunity to evaluate the timing and magnitude of Late Quaternary deformation at the northern edge of the rotated western Transverse Ranges. Three flights of well-preserved fluvial strath terraces are exposed along a 9.3 km stretch of the Santa Cruz Creek drainage in the eastern Santa Ynez Valley. Fluvial deposits that make up the terrace treads have been lifted 18.5 m (T-1) to 110m (T-3) above the active channel and are unpaired across the drainage. The terrace surfaces are underlain by a thin 0.6m-6m veneer of fluvial deposits resting on an erosional surface cut into the underlying Pleistocene Paso Robles Formation. GPS profiles (accurate to .1m) of the terrace surfaces and the Santa Cruz Creek stream profile show evidence of Late Quaternary deformation. The T1 surface (lowest terrace surface) is offset vertically 3.5m across the Los Alamos/Baseline fault, the T2 surface is offset 7m vertically across this feature, and the T2 and T3 terraces terminate abruptly at the Little Pine Fault indicating Quaternary displacement across these structures. Optically Stimulated Luminescence (OSL) dating is being used to date the terraces in order to calculate rates of uplift and deformation.

  11. 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.

  12. 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

  13. Community structures of actively growing bacteria shift along a north-south transect in the western North Pacific.

    PubMed

    Taniguchi, Akito; Hamasaki, Koji

    2008-04-01

    Bacterial community structures and their activities in the ocean are tightly coupled with organic matter fluxes and thus control ocean biogeochemical cycles. Bromodeoxyuridine (BrdU), halogenated nucleoside and thymidine analogue, has been recently used to monitor actively growing bacteria (AGB) in natural environments. We labelled DNA of proliferating cells in seawater bacterial assemblages with BrdU and determined community structures of the bacteria that were possible key species in mediating biochemical reactions in the ocean. Surface seawater samples were collected along a north-south transect in the North Pacific in October 2003 and subjected to BrdU magnetic beads immunocapture and PCR-DGGE (BUMP-DGGE) analysis. Change of BrdU-incorporated community structures reflected the change of water masses along a north-south transect from subarctic to subtropical gyres in the North Pacific. We identified 25 bands referred to AGB as BrdU-incorporated phylotypes, belonging to Alphaproteobacteria (5 bands), Betaproteobacteria (1 band), Gammaproteobacteria (4 bands), Cytophaga-Flavobacterium-Bacteroides (CFB) group bacteria (5 bands), Gram-positive bacteria (6 bands), and Cyanobacteria (4 bands). BrdU-incorporated phylotypes belonging to Vibrionales, Alteromonadales and Gram-positive bacteria appeared only at sampling stations in a subtropical gyre, while those belonging to Roseobacter-related bacteria and CFB group bacteria appeared at the stations in both subarctic and subtropical gyres. Our result revealed phylogenetic affiliation of AGB and their dynamic change along with north-south environmental gradients in open oceans. Different species of AGB utilize different amount and kinds of substrates, which can affect the change of organic matter fluxes along transect.

  14. The mouse prostaglandin E receptor EP2 subtype: cloning, expression, and northern blot analysis.

    PubMed

    Katsuyama, M; Nishigaki, N; Sugimoto, Y; Morimoto, K; Negishi, M; Narumiya, S; Ichikawa, A

    1995-09-25

    A functional cDNA clone for the mouse prostaglandin (PG) E receptor EP2 subtype was isolated from a mouse cDNA library. The mouse EP2 receptor consists of 362 amino acid residues with seven putative transmembrane domains. [3H]PGE2 bound specifically to the membrane of Chinese hamster ovary cells stably expressing the cloned receptor. This binding was displaced by unlabeled prostanoids in the order of PGE2 = PGE1 > iloprost, a stable PGI2 agonist > PGF2 alpha > PGD2. Binding was also inhibited by butaprost (an EP2 agonist) and to a lesser extent by M&B 28767 (an EP3 agonist), but not by sulprostone (an EP1 and EP3 agonist) or SC-19220 (an EP1 antagonist). PGE2 and butaprost increased the cAMP level in the Chinese hamster ovary cells in a concentration-dependent manner. Northern blot analysis revealed that EP2 mRNA is expressed most abundantly in the uterus, followed by the spleen, lung, thymus, ileum, liver, and stomach.

  15. Diagnosis of cutaneous tuberculosis in biopsy specimens by PCR and southern blotting.

    PubMed Central

    Quirós, E; Maroto, M C; Bettinardi, A; González, I; Piédrola, G

    1996-01-01

    AIMS: To evaluate the use of a gene amplification and hybridisation method for detecting mycobacterial nucleic acid as a possible diagnostic method for cutaneous tuberculosis infection. METHODS: Biopsy specimens from 20 patients with various skin conditions of possible tuberculous aetiology were studied. Six patients had ulcerative nodules, seven lupiform lesions, two non-necrotic granulomas, one scrofulous lichen, one impetigo, one erythematosus lesions, one warty lesions, and one suspected tuberculous lipoma. Biopsy specimens were stained using Ziehl-Neelsen stain and cultured in Lowenstein-Jensen medium. DNA was extracted and then amplified by PCR using primers specific for the Mycobacterium tuberculosis complex. Specificity was confirmed by Southern blotting. RESULTS: Of the specimens, 30% were positive for mycobacteria on staining with Ziehl-Neelsen stain, 60% were culture positive and 85% PCR positive. Only 35.2% of specimens were positive with all three techniques. A further 32.5% were both culture and PCR positive. All PCR negative samples were also negative when cultured or stained with Ziehl-Neelsen stain. Of the PCR positive specimens, 29.4% were negative when cultured or stained. CONCLUSIONS: PCR, using suitable primers, is an efficient and sensitive method for the diagnosis of cutaneous tuberculosis. PMID:8944606

  16. A simple dot-blot-Sirius red-based assay for collagen quantification.

    PubMed

    Rodríguez-Rodríguez, Pilar; Arribas, Silvia M; de Pablo, Angel Luis López; González, M Carmen; Abderrahim, Fatima; Condezo-Hoyos, Luis

    2013-08-01

    The assessment of collagen content in tissues is important in biomedical research, since this protein is altered in numerous diseases. Hydroxyproline and Sirius red based assays are the most common methods for collagen quantification. However, these procedures have some pitfalls, such as the requirement of oxygen-free medium or expensive equipment and large sample size or being unsuitable for hydrolyzed collagen, respectively. Our objective was to develop a specific, versatile, and user-friendly quantitative method applicable to small tissue samples and extracts obtained from elastin purification, therefore, suitable for simultaneous quantification of elastin. This method is based on the binding of Sirius red to collagen present in a sample immobilized on a PVDF membrane, as in the dot-blot technique, and quantified by a scanner and image analysis software. Sample loading, Sirius red concentration, temperature and incubation time, type of standard substance, albumin interference, and quantification time are optimized. The method enabled the quantification of (1) intact collagen in several rat tissue homogenates, including small resistance-sized arteries, (2) partially hydrolyzed collagen obtained from NaOH extracts, compatible with elastin purification, and (3) for the detection of differences in collagen content between hypertensive and normotensive rats. We conclude that the developed technique can be widely used since it is versatile (quantifies intact and hydrolyzed collagen), requires small sample volumes, is user-friendly (low-cost, easy to use, minimum toxic materials, and reduced time of test), and is specific (minimal interference with serum albumin).

  17. Genotype-specific RNA probes for direct identification of wild polioviruses by blot hybridization.

    PubMed Central

    De, L; Yang, C F; Da Silva, E; Boshell, J; Cáceres, P; Gómez, J R; Pallansch, M; Kew, O

    1997-01-01

    We have developed RNA probes for the direct identification of wild poliovirus isolates by blot hybridization. The probes are complementary to sequences of the first 30 to 32 codons of VP1, which evolve more extensively (approximately 1.5-fold) than the rest of VP1. To illustrate our general approach, we describe the design of probes specific to each of four major genotypes recently endemic (1981 to 1991) to the Americas: Andean type 1, Brazil type 1, Brazil type 3, and Central America-Mexico type 3. A wild isolate of each genotype was selected according to molecular and epidemiologic criteria to be representative of the principal lineages in circulation. Variable VP1 sequences of the representative isolates were amplified by the reverse transcriptase PCR and were inserted into a plasmid vector containing a T7 promoter. The in vitro transcripts, labeled with digoxigenin, served as probes. These formed stable hybrids only with RNAs of isolates of the corresponding genotypes. Hybrids were detected by a sensitive chemiluminescence assay, capable under normal diagnostic conditions of detecting specific wild poliovirus sequences in samples containing up to a 100-fold excess of Sabin vaccine strain-related sequences of the same serotype. PMID:9350743

  18. 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

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

    PubMed

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

    2015-10-01

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

  20. Evaluation of an enzyme-linked immunoelectrotransfer blot test for the confirmatory serodiagnosis of human toxocariasis.

    PubMed

    Roldán, William H; Espinoza, Yrma A

    2009-05-01

    To improve the serodiagnosis of human toxocariasis, a sensitive and specific enzyme-linked immunoelectrotransfer blot (EITB-IgG) test was developed and evaluated using Toxocara canislarvae excretory-secretory antigens for detecting anti-Toxocara IgG antibodies. The EITB-IgG profile of toxocariasis was characterized by comparing 27 sera from patients with toxocariasis, 110 sera from healthy subjects and 186 sera from patients with other helminth diseases (ascariasis, ancylostomiasis, trichuriasis, enterobiasis, strongyloidiasis, hymenolepiasis, diphyllobothriasis, taeniasis, cysticercosis, hydatidosis and fascioliasis). Antigenic bands of 24, 28, 30, 35, 56, 117, 136 and 152 kDa were predominantly recognized in sera from all patients with toxocariasis. However, only bands of 24-35 kDa were highly specific for Toxocara infection (98.3%), whereas other antigenic bands observed displayed cross-reactivity. Additionally, when the results of the EITB-IgG test were compared to those of the ELISA-IgG test, a 100% concordance was observed for positive results in human toxocariasis cases. The concordance for negative results between the two tests for healthy subjects and patients with other helminth diseases were 96.3% and 53.7%, respectively, showing that the EITB-IgG test has a higher specificity than ELISA. In conclusion, the EITB-IgG test is a very useful tool to confirm the serological diagnosis of human toxocariasis.

  1. Renaturation of blotted allergens increases the sensitivity of specific IgE detection.

    PubMed

    Muro, M D; Fernández, C; Moneo, I

    1996-01-01

    Several authors have demonstrated that renaturation is an essential step for the appropriate recognition of blotted proteins. The use of nonionic detergents has been described as a useful alternative to enhance the antigenicity in immunoblotting, although elution from proteins by detergents has been observed. To measure the influence of different factors on the sensitivity of specific IgE by immunoblotting, we used twenty human sera from atopic patients who were allergic or nonallergic to a common, reliable allergen (grass pollen mixture). The use of Nonidet-P40 was found to be a useful alternative for the renaturation of the allergens. No elution from the membrane was found when employing this detergent, even at high concentrations (3%), and its use gave better sensitivity than methanol. On the other hand, we detected that methanol possessed renaturing properties. A transfer method using diffusion instead of electric transfer gave the best results and two membranes could be obtained from each gel. Using this method, we found that after NP-40 incubation of the membrane, the use of bovine albumin could be omitted as blocking agent and that its use had even deleterious effects.

  2. Trypanosoma cruzi: variability of stocks from Colombia determined by molecular karyotype and minicircle Southern blot analysis.

    PubMed

    Triana, Omar; Ortiz, Sylvia; Dujardin, Jean-Claude; Solari, Aldo

    2006-05-01

    Nineteen Trypanosoma cruzi stocks, most of them of wild origin, and four Trypanosoma rangeli stocks from Colombia were analysed by molecular karyotype analysis with cloned DNA cruzipain as the probe. Another 27 cloned stocks of T. cruzi from different geographic areas of South America were used as reference for T. cruzi lineages. Phenetic analysis of chromosome size polymorphism demonstrated a great variability of Colombian T. cruzi stocks, suggesting that most belong to lineage I, although two of them belong to lineage II. The 2 lineage II T. cruzi, 17 T. cruzi lineage I, and 3 T. rangeli stocks from Colombia were studied further by Southern blot analysis with a panel of kinetoplast DNA minicircle probes. Hybridisation results indicate that the two T. cruzi II stocks are genetically distant from each other and from T. cruzi lineages IIb, IId, and IIe from Chile. Finally, T. cruzi minicircle probes do not cross-hybridise in any stringency condition tested with T. rangeli minicircles, a clear indication that these parasites can be easily distinguished by this method.

  3. 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.

  4. 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.

  5. 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

  6. 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.

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

    PubMed 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

  8. 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

  9. Seasonal variations in active dispersal of natural populations of Triatoma infestans in rural north-western Argentina

    PubMed Central

    VAZQUEZ-PROKOPEC, G. M.; CEBALLOS, L.A.; MARCET, P. L.; CECERE, M. C.; CARDINAL, M. V.; KITRON, U.; GÜRTLER, R. E.

    2007-01-01

    The flight dispersal of Triatoma infestans Klug (Hemiptera: Reduviidae) is one of the main mechanisms determining community re-infestation after control interventions. An empirical model of flight initiation coupled with data from a longitudinal study predicted that the flight dispersal of T. infestans would peak in summer. To test this prediction, longitudinal light trap collections were conducted during 3-8 nights in March (late summer), July (winter) and November (spring) 2003, and in March 2004 in a rural community in north-west Argentina. Following each light-trapping collection date, all peridomestic sites around light traps were inspected to assess the relative abundance and nutritional status of T. infestans at each site. A total of 21 adult and five nymph T. infestans, six Triatoma guasayana Wygodzinsky & Abalos, and nine Triatoma garciabesi Carcavallo et al. were collected in 96 light-trapping nights, whereas 696 T. infestans were collected from the peridomestic sites that surrounded the light traps. The arrival of T. infestans in the light traps occurred in 64% of catch stations and peaked in the summer surveys (10-14 bugs) compared with spring and winter surveys. When winds were < 5 km/h, the arrival of adult T. infestans at the light traps was significantly associated with maximum temperature and relative humidity. This is the first field report of seasonal variations in the flight dispersal activity of T. infestans. PMID:17044877

  10. Seasonal variations in active dispersal of natural populations of Triatoma infestans in rural north-western Argentina.

    PubMed

    Vazquez-Prokopec, G M; Ceballos, L A; Marcet, P L; Cecere, M C; Cardinal, M V; Kitron, U; Gürtler, R E

    2006-09-01

    The flight dispersal of Triatoma infestans Klug (Hemiptera: Reduviidae) is one of the main mechanisms determining community re-infestation after control interventions. An empirical model of flight initiation coupled with data from a longitudinal study predicted that the flight dispersal of T. infestans would peak in summer. To test this prediction, longitudinal light trap collections were conducted during 3-8 nights in March (late summer), July (winter) and November (spring) 2003, and in March 2004 in a rural community in north-west Argentina. Following each light-trapping collection date, all peridomestic sites around light traps were inspected to assess the relative abundance and nutritional status of T. infestans at each site. A total of 21 adult and five nymph T. infestans, six Triatoma guasayana Wygodzinsky & Abalos, and nine Triatoma garciabesi Carcavallo et al. were collected in 96 light-trapping nights, whereas 696 T. infestans were collected from the peridomestic sites that surrounded the light traps. The arrival of T. infestans in the light traps occurred in 64% of catch stations and peaked in the summer surveys (10-14 bugs) compared with spring and winter surveys. When winds were < 5 km/h, the arrival of adult T. infestans at the light traps was significantly associated with maximum temperature and relative humidity. This is the first field report of seasonal variations in the flight dispersal activity of T. infestans.

  11. Soils and geomorphic evolution of bedrock facets on a tectonically active mountain front, western Sangre de Cristo Mountains, New Mexico

    NASA Astrophysics Data System (ADS)

    Menges, Christopher M.

    1990-09-01

    Soil profiles, colluvial stratigraphy, and detailed hillslope morphology are key elements used for geomorphic interpretations of the form and long-term evolution of triangular facets on a 1200 m high, tectonically active mountain front. The facets are developed on Precambrian gneisses and Tertiary volcanic and plutonic rocks along a complexly segmented, active normal-fault zone in the Rio Grande rift of northern New Mexico. The detailed morphologies of 20- to 350 m high facets are defined by statistical and time-series analyses of 40 field transects that were keyed to observations of colluvium, bedrock, microtopography, and vegetation. The undissected parts of most facets are transport-limited hillslopes mantled with varying thicknesses (0.1 to > 1 m thick) of sand and gravel colluvium between generally sparse (≤10-30%) bedrock outcrops. Facet soils range from (a) thin (≤ 0.2 m) weakly developed soils with cumulic silty A or transitional A/B epipedons above Cox horizons in bedrock or colluvium, to (b) deep (≥0.5-1 m) moderately to strongly developed profiles containing thick cambic (Bw) and/or argillic (Bt) horizons that commonly extend into highly weathered saprolitic bedrock. The presence of strongly weathered profiles and thick colluvium suggests that rates of colluvial transport and hillslope erosion are less than or equal to rates of soil development over at least a large part of the Holocene. The catenary variation of soils and colluvium on selected facet transects indicate that the degree of soil development generally increases and the thickness of colluvium decreases upslope on most facets. This overall pattern is commonly disrupted on large facet hillslopes by irregular secondary soil variations linked to intermediate-scale (20-60 + m long) concave slope elements. These features are interpreted to reflect discontinuous transport and erosion of colluvium down-slope below bedrock outcrops. The degree of weathering in subsurface bedrock commonly

  12. 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.

  13. 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

  14. 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

  15. 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...

  16. 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

  17. Two-Phase Contiguous Supported Lipid Bilayer Model for Membrane Rafts via Polymer Blotting and Stenciling.

    PubMed

    Richards, Mark J; Daniel, Susan

    2017-02-07

    The supported lipid bilayer has been portrayed as a useful model of the cell membrane compatible with many biophysical tools and techniques that demonstrate its appeal in learning about the basic features of the plasma membrane. However, some of its potential has yet to be realized, particularly in the area of bilayer patterning and phase/composition heterogeneity. In this work, we generate contiguous bilayer patterns as a model system that captures the general features of membrane domains and lipid rafts. Micropatterned polymer templates of two types are investigated for generating patterned bilayer formation: polymer blotting and polymer lift-off stenciling. While these approaches have been used previously to create bilayer arrays by corralling bilayers patches with various types of boundaries impenetrable to bilayer diffusion, unique to the methods presented here, there are no physical barriers to diffusion. In this work, interfaces between contiguous lipid phases define the pattern shapes, with continuity between them allowing transfer of membrane-bound biomolecules between the phases. We examine effectors of membrane domain stability including temperature and cholesterol content to investigate domain dynamics. Contiguous patterning of supported bilayers as a model of lipid rafts expands the application of the SLB to an area with current appeal and brings with it a useful toolset for characterization and analysis. These combined tools should be helpful to researchers investigating lipid raft dynamics and function and biomolecule partitioning studies. Additionally, this patterning technique may be useful for applications such as bioseparations that exploit differences in lipid phase partitioning or creation of membranes that bind species like viruses preferentially at lipid phase boundaries, to name a few.

  18. 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

  19. The character and reactivation history of the southern extension of the seismically active Clarendon Linden Fault System, western New York State

    NASA Astrophysics Data System (ADS)

    Jacobi, Robert D.; Fountain, John

    2002-08-01

    Integration of 11 types of data sets enabled us to determine the location, character and fault history of the southern extension of the Clarendon-Linden Fault System (CLF) in southwestern New York State. The data sets utilized include detailed stratigraphic and fracture measurements at more than 1000 sites, soil gas anomalies, seismic reflection profiles, well logs and lineaments on air photos, topographic maps, Landsat and SLAR images. The seismically active CLF consists of as many as 10 parallel, segmented faults across the fault system. The fault segments are truncated by NW-striking cross-strike discontinuities (CSDs). The faults of the CLF and intersecting CSDs form fault blocks that have semi-independent subsidence and uplift histories. East-dipping reflectors in the Precambrian basement indicate the southward continuation of thrusts of the intra-Grenvillian Elzevir-Frontenac Boundary Zone. These thrusts were reactivated during Iapetan rifting as normal (listric) growth faults. In Ordovician Black River to Trenton time, the southern CLF segments experienced a second phase of growth fault activity, with faults displaying a cumulative stratigraphic throw of as much as ˜170 m. Thrusting on the same east-dipping Precambrian reflectors typified the CLF in Taconic (post-Trenton) times. Detailed comparisons among the fault segments show that the fault activity in Silurian and Devonian times generally alternated between the western and central main faults. In Late Devonian time, the fault motion reversed from down-on-the-east to down-on-the-west about the time the Appalachian Basin axis passed across the CLF in its westward migration. The deep Precambrian faults of the CLF were thus reactivated as the Appalachian Basin developed in Acadian times. Finally, the CLF thrust fault imaged on seismic line CLF-1 offsets all bedrock (Devonian) units; thus, significant motion occurred along this fault during Late Acadian, or more likely, Alleghanian time.

  20. A rapid chemiluminescent slot blot immunoassay for the detection and quantification of Clostridium botulinum neurotoxin type E, in cultures.

    PubMed

    Cadieux, Brigitte; Blanchfield, Burke; Smith, James P; Austin, John W

    2005-05-01

    A simple, rapid, cost-effective in vitro slot blot immunoassay was developed for the detection and quantification of botulinum neurotoxin type E (BoNT/E) in cultures. Culture supernatants of 36 strains of clostridia, including 12 strains of Clostridium botulinum type E, 12 strains of other C. botulinum neurotoxin serotypes, and 12 strains of other clostridial species were tested. Samples containing BoNT/E were detected using affinity-purified polyclonal rabbit antisera prepared against BoNT/E with subsequent detection of secondary antibodies using chemiluminescence. All strains of C. botulinum type E tested positive, while all non C. botulinum type E strains tested negative. The sensitivity of the slot blot immunoassay for detection of BoNT/E was approximately four mouse lethal doses (MLD). The intensity of chemiluminescence was directly correlated with the concentration of BoNT/E up to 128 MLD, allowing quantification of BoNT/E between 4 and 128 MLD. The slot blot immunoassay was compared to the mouse bioassay for detection of BoNT/E using cultures derived from fish samples inoculated with C. botulinum type E, and cultures derived from naturally contaminated environmental samples. A total of 120 primary enrichment cultures derived from fish samples, of which 103 were inoculated with C. botulinum type E, and 17 were uninoculated controls, were assayed. Of the 103 primary enrichment cultures derived from inoculated fish samples, all were positive by mouse bioassay, while 94 were also positive by slot blot immunoassay, resulting in a 7.5% false-negative rate. All 17 primary enrichment cultures derived from the uninoculated fish samples were negative by both mouse bioassay and slot blot immunoassay. A total of twenty-six primary enrichment cultures derived from environmental samples were tested by mouse bioassay and slot blot immunoassay. Of 13 primary enrichment cultures positive by mouse bioassay, 12 were also positive by slot blot immunoassay, resulting in a 3

  1. 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.

  2. Double-blotting: a solution to the problem of non-specific binding of secondary antibodies in immunoblotting procedures.

    PubMed

    Lasne, F

    2001-07-01

    "Double-blotting" (DB) was developed to overcome the problem of non-specific binding of secondary antibodies in immunoblotting (IB). After it had been probed by the primary antibody, the membrane with the blotted proteins was assembled with a second blank membrane and submitted to a second blotting under acidic conditions. The primary antibody molecules were thus desorbed from their corresponding antigen and transferred onto the second membrane, whereas the antigen and the interfering proteins remained bound to the first one. The second membrane could then be probed by the secondary antibodies without the risk of non-specific binding. This method was developed for the study of erythropoietin (EPO) in concentrated urine since a strong non-specific binding of biotinylated secondary antibodies to some urinary proteins had been observed using classical IB protocols.

  3. Double-blotting: a solution to the problem of nonspecific binding of secondary antibodies in immunoblotting procedures.

    PubMed

    Lasne, Françoise

    2003-05-01

    "Double-blotting" (DB) has been developed to overcome the problem of nonspecific binding of secondary antibodies in immunoblotting (IB). After it has been probed by the primary antibody, the membrane with the blotted proteins is assembled with a second blank membrane and submitted to a second blotting under acidic conditions. The primary antibody molecules are thus desorbed from their corresponding antigen and transferred onto the second membrane, whereas the antigen and the interfering proteins remain bound to the first one. The second membrane can then be probed by the secondary antibodies without the risk of nonspecific binding. This method has been developed for the study of erythropoietin (EPO) in concentrated urine since a strong nonspecific binding of biotinylated secondary antibodies to some urinary proteins is observed using classical IB protocols. However, its concept makes it usable in other applications that come up against this kind of problem.

  4. Detection of enterotoxigenic Clostridium perfringens in spices used in Mexico by dot blotting using a DNA probe.

    PubMed

    Rodríguez-Romo, L A; Heredia, N L; Labbé, R G; García-Alvarado, J S

    1998-02-01

    Several reports on the microbiology of spices and herbs indicate the presence of Clostridium perfringens, a spore-forming foodborne pathogen responsible for gastrointestinal disease. In the present study, a total of 380 samples of spices and herbs (cumin seed, black pepper, oregano, garlic powder, and bay leaves) widely used in Mexico were analyzed for the presence of C. perfringens, and the enterotoxigenicity of the isolates was determined by a dot-blot technique using an enterotoxin degoxigenin-labeled DNA probe. C. perfringens counts varied from <100 to 433 CFU/g in garlic powder, from <100 to 200 CFU/g in black pepper, from <100 to 433 CFU/g in cumin seed, from <100 to 340 CFU/g in oregano, and from < 100 to 450 CFU/g in bay leaves. The dot-blot technique detected the enterotoxin gene in 8 (4.25%) of 188 confirmed isolates of C. perfringens. dot-blot.

  5. 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

  6. Identification of a diverse synthetic abietane diterpenoid library for anticancer activity.

    PubMed

    Xu, Hongtao; Liu, Lili; Fan, Xiaoting; Zhang, Guanjun; Li, Yuanchao; Jiang, Biao

    2017-02-01

    A diverse natural-product-like synthetic abietane diterpenoid library contains about 56 compounds were obtained, and evaluated for their potential in vitro cytotoxic or antitumor activity against A549, PC-3 and SKOV-3 tumor cell lines by SRB assay. Treatment of A549 cells with the most potent compound ketone 19 showed induction of apoptosis, as revealed by JC-1 mitochondrial membrane potential staining, TUNNEL assay, western blotting analysis and flow cytometry assay.

  7. Hypoxia-Inducible Factor-1alpha and MAPK Co-Regulate Activation of Hepatic Stellate Cells upon Hypoxia Stimulation

    PubMed Central

    Guan, Fei; Xiao, Yan; Deng, Jing; Chen, Huoying; Chen, Xiaolin; Li, Jianrong; Huang, Hanju; Shi, Chunwei

    2013-01-01

    Background Hepatic stellate cell (HSC) plays a key role in pathogenesis of liver fibrosis. During liver injury, hypoxia in local micro-environment is inevitable. Hif-1α is the key transcriptional regulation factor that induces cell’s adaptive responses to hypoxia. Recently, it was reported that MAPK is involved in regulation of Hif-1α activity. Aims To explore whether Hif-1α regulates HSC activation upon hypoxia, and whether MAPK affects Hif-1α-regulated signaling cascades, thus providing new targets for preventing liver fibrosis. Methods Hif-1α expression in livers of Schistosomajaponicum infected BALB/c mice was detected with western blot and immunohistochemistry. A rat cell line of HSC, HSC-T6, was cultured in 1% oxygen. HSC activation, including F-actin reorganization, increase of vimentin and α-SMA, was detected with western blot or immunocytochemistry. Cells were transfected with specific siRNA to Hif-1α, expression of activation markers, transcription of fibrosis-promoting cytokines, secretion of collagen I were detected with western blot, Real Time PCR and ELISA. Lysate from HSC-T6 cells pretreated with PD98059, a specific MEK1 pharmacological inhibitor, was subjected to detect Hif-1α ubiquitination and nuclear translocation with western blot and immunoprecipitation. Results and Conclusions Hif-1α apparently increased in liver tissues of Schistosomajaponicum infected mice. 1% O2 induced F-actin reorganization, increase of Hif-1α, vimentin and α-SMA in HSC-T6 cells. Hif-1α Knockdown inhibited HSC-T6 activation, transcription of IL-6, TGF-β and CTGF and secretion of collagen I from HSC-T6 cells upon hypoxia. Inhibition of MAPK phosphorylation enhanced Hif-1α ubiquitination, and inhibited Hif-1α translocation into nucleus. Conclusively, Hif-1α and MAPK participate in HSC activation upon hypoxia. PMID:24040163

  8. Immunofluorescence assay reactivity patterns of serum samples presenting indeterminate Western blot results for antibodies to HIV-1 and HTLV-I/II in Cordoba, Argentina.

    PubMed

    Gastaldello, R; Gallego, S; Isa, M B; Maturano, E; Sileoni, S; Nates, S; Medeot, S

    2001-01-01

    Serum samples (n: 110) from blood donors and high risk individuals from Cordoba, Argentina with indeterminate HIV-1 and HTLV-I/II Wb profiles were studied for specific antibodies to HTLV-I/II and HIV-1 by indirect immunofluorescence assay (IFA) and for the presence or absence of HIV-1 and HTLV-I/II specific bands by Wb. This study was carried out in order to characterize their putative reactions with HIV-1 and HTLV-I/II proteins and to resolve the retrovirus infection status of these individuals. Results indicated that blood donors sera displaying indeterminate HIV-1 or HTLV-I/II Wb patterns were not immunoreactive to HTLV-I/II and HIV-1 on IFA. However, a high rate of indeterminate HIV-1 and HTLV-I/II Wb samples from high risk individuals had positive HTLV-I/II and HIV-1 IFA results respectively. Our study supports the growing evidence that HTLV-HIV indeterminate seroreactivity in low risk population is due to a cross reaction against nonviral antigens, and in high risk populations the indeterminate samples show serological cross-recognition between HIV-1 proteins and HTLV-I/II proteins on Wb. These results point out the necessity to investigate the HTLV-I/II reactivity in indeterminate HIV-1 samples and vice versa in order to confirm the diagnosis. Finally, this study shows the potential usefulness of IFA in elucidating the status of HIV-1 and HTLV-I/II infection of individuals with indeterminate Wb profiles, thus enabling resolution of retrovirus infection status.

  9. Long-term serological follow-up of blood donors with an HTLV-indeterminate western blot: antibody profile of seroconverters and individuals with false reactions.

    PubMed

    Martins, Marina Lobato; Santos, Ana Carolina da Silva; Namen-Lopes, Maria Sueli; Barbosa-Stancioli, Edel Figueiredo; Utsch, Denise Gonçalves; Carneiro-Proietti, Anna Bárbara de F

    2010-10-01

    The high proportion of indeterminate results of the screening test for human T-lymphotropic virus (HTLV) infection has been a challenge worldwide. In this study, 60 persons with seroindeterminate results for HTLV were followed until their serological status was defined. At least two independent serological tests (EIA and WB) from sequential samples were performed at an average interval of 4.4 years, totaling 141 serum samples tested. Seroconversion occurred in 12 individuals (reactive by EIA, positive by WB and PCR), and 48 were classified as false reactions (non-reactive EIA and negative PCR, but indeterminate WB). The seroconverter group had epidemiological features similar to those seen in HTLV-1 carriers, and the average time of follow-up for seroconversion was 4 years. In the group with false reactions, the most frequent indeterminate WB pattern in the samples was the presence of p24 alone. This pattern was absent in the seroconverter group, suggesting that p24 alone is an indicator of false reactivity. In contrast, the presence of p19 and p24 seems to be an indicator of true reactivity, since this pattern was frequent (66.7%) among the seroconverters and much less common (10.4% of the first samples) among the individuals with false reactions (P = 0.0001). Thus, HTLV infection may be suspected when reactivity to p19 and p24 is observed. Individuals with an indeterminate WB pattern should be followed-up and retested. The improvement of the HTLV algorithm screening of blood donors has been necessary to reduce inconclusive results and to avoid unnecessary follow-up to define the status of infection.

  10. Use of Sera from Humans and Dolphins with Lacaziosis and Sera from Experimentally Infected Mice for Western Blot Analyses of Lacazia loboi Antigens▿

    PubMed Central

    Mendoza, Leonel; Belone, Andréa F. F.; Vilela, Raquel; Rehtanz, Manuela; Bossart, Gregory D.; Reif, John S.; Fair, Patricia A.; Durden, Wendy N.; St. Leger, Judy; Travassos, Luiz R.; Rosa, Patricia S.

    2008-01-01

    Antibodies in the sera of patients with lacaziosis recognized an ∼193-kDa antigen and other Lacazia loboi antigens. Paracoccidioides brasiliensis gp43 antigen was detected by all evaluated sera, but they failed to detect a protein with the same molecular mass in L. loboi extracts. This study is the first to examine the humoral response to L. loboi antigens by using multiple host sera. PMID:17959822

  11. 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...

  12. 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.

  13. Male-associated polypeptide (MAP) expression in different compartments of the reproductive system of the mussel Mytilus galloprovincialis: immunocytochemical and western blot study.

    PubMed

    Torrado, M; Mikhailov, A T

    1998-12-01

    Mytilus mussels are characterized by annually repeated reproduction which is associated with subsequent growth, morphogenesis, breakdown and redevelopment of the gonad and reproductive tract into mantle mesenchyme. We present a description of the expression of the male-associated polypeptide (MAP; see Mikhailov et al. 1995) in different compartments of the male reproductive system as well as in mantle gonad-supporting tissue. MAP is expressed in both gonad and mantle structures in dynamic patterns that show a substantial overlap in terms of dependence on the stage of gonad development/involution. In general, the total MAP concentration directly correlates with the volume of gonad tubule/duct structures but inversely correlates with mantle connective tissue cell fraction. A maximum of MAP expression is reached in the fully ripe male gonad. MAP is localized around gonad tubules/ducts, in the gonoduct epithelium, membranes of follicle-like structures as well as in the extracellular fiber-like structures of the mantle. However, we also demonstrate unique sites of MAP accumulation in the lumen of gonad follicle-like tubules and in ductal fluid. The latter is characterized by a very high MAP concentration. MAP is also detected in sperm-containing cell suspension obtained by gonad biopsy which we interpret as a result of the adsorption of MAP on mature spermatozoa. The results obtained should be taken into consideration in the interpretation of possible MAP functions since they seem to point to MAP as a major component of ductal (seminal) fluid of the male reproductive tract. It is likely that MAP is able to complement the processes of sperm terminal differentiation and maturation. In addition, we demonstrate that the male-predominant character of MAP expression is restricted by gonad-containing tissues (i.e., mantle and visceral mass) only, although the polypeptide is also detected in other somatic organs in both males and females.

  14. 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…

  15. SorLA and CLC:CLF-1-dependent Downregulation of CNTFRα as Demonstrated by Western Blotting, Inhibition of Lysosomal Enzymes, and Immunocytochemistry.

    PubMed

    Larsen, Jakob V; Petersen, Claus M

    2017-01-06

    The heterodimeric cytokine Cardiotrophin-like Cytokine:Cytokine-like Factor-1 (CLC:CLF-1) targets the glycosylphosphatidylinositol (GPI)-anchored CNTFRα to form a trimeric complex that subsequently recruits glycoprotein 130/Leukemia Inhibitory Factor Receptor-β (gp130/LIFRβ) for signaling. Both CLC and CNTFRα are necessary for signaling but so far CLF-1 has only been known as a putative facilitator of CLC secretion. However, it has recently been shown that CLF-1 contains three binding sites: one for CLC; one for CNTFRα (that may promote assembly of the trimeric complex); and one for the endocytic receptor sorLA. The latter site provides high affinity binding of CLF-1, CLC:CLF-1, as well as the trimeric (CLC:CLF-1:CNTFRα) complex to sorLA, and in sorLA-expressing cells the soluble ligands CLF-1 and CLC:CLF-1 are rapidly taken up and internalized. In cells co-expressing CNTFRα and sorLA, CNTFRα first binds CLC:CLF-1 to form a membrane-associated trimeric complex, but it also connects to sorLA via the free sorLA-binding site in CLF-1. As a result, CNTFRα, which has no capacity for endocytosis on its own, is tugged along and internalized by the sorLA-mediated endocytosis of CLC:CLF-1. The present protocol describes the experimental procedures used to demonstrate i) the sorLA-mediated and CLC:CLF-1-dependent downregulation of surface-membrane CNTFRα expression; ii) sorLA-mediated endocytosis and lysosomal targeting of CNTFRα; and iii) the lowered cellular response to CLC:CLF-1-stimulation upon sorLA-mediated downregulation of CNTFRα.

  16. 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

  17. 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.

  18. 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

  19. 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.

  20. 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

  1. Wood Bark Smoke Induces Lung and Pleural Plasminogen Activator Inhibitor 1 and Stabilizes Its mRNA in Porcine Lung Cells

    DTIC Science & Technology

    2011-08-01

    in situ. Plasminogen activator inhibitor 1 was measured in bronchoalveolar lavage fluids by Western blotting. Induction of PAI-1 was determined at the...As measured in bronchoalveolar lavage (BAL) fluid, this defect in fibrinolysis is mainly attributable to overexpression of plasminogen activator...monitoring Pigs were monitored for 48 h. The following variables were measured : number of smoke breaths received, volume of smoke received, peak carboxy

  2. Preference for Western diet coadapts in High Runner mice and affects voluntary exercise and spontaneous physical activity in a genotype-dependent manner.

    PubMed

    Acosta, Wendy; Meek, Thomas H; Schutz, Heidi; Dlugosz, Elizabeth M; Garland, Theodore

    2017-02-01

    Do animals evolve (coadapt) to choose diets that positively affect their performance abilities? We addressed this question from a microevolutionary perspective by examining preference for Western diet (WD: high in fat and sugar, but lower in protein) versus standard rodent chow in adults of both sexes from 4 lines of mice selectively bred for high levels of voluntary wheel running (High Runner or HR lines) and 4 non-selected control (C) lines. We also assessed whether food preference or substitution affects physical activity (wheel running and/or spontaneous physical activity [SPA] in the attached home cages). In experiment 1 (generation 56), mice were given 6days of wheel acclimation (as is used routinely to pick breeders in the selection experiment) prior to a 2-day food choice trial. In experiment 2 (generation 56), 17days of wheel acclimation allowed mice to reach a stable level of daily running, followed by a 7-day food-choice trial. In experiment 3 (generation 58), mice had 6days of wheel acclimation with standard chow, after which half were switched to WD for two days. In experiment 1, WD was highly preferred by all mice, with somewhat greater preference in male C mice. In experiment 2, wheel running increased and SPA decreased continuously for the first 14days of adult wheel testing, followed by 3-day plateaus in both. During the subsequent 7-day food choice trial, HR mice of both sexes preferred WD significantly more than did C mice; moreover, wheel running increased in all groups except males from C lines, with the increase being significantly greater in HR than C, while SPA declined further in all groups. In experiment 3, the effect of being switched to WD depended on both linetype and sex. On standard chow, only HR females showed a significant change in wheel running during nights 7+8, increasing by 10%. In contrast, when switched to WD, C females (+28%), HR females (+33%), and HR males (+10%) all significantly increased their daily wheel

  3. Detection of X chromosome aneuploidy using Southern blot analysis during routine population-based screening for fragile X syndrome.

    PubMed

    Adir, Vardit; Shahak, Elena; Dar, Hanna; Borochowitz, Zvi U

    2003-01-01

    We report herein two cases where detection of X chromosome aneuploidy (cytogenetically proved 45,X/46XX and 47,XXX) was made possible by molecular diagnosis during population-based carrier screening for Fragile X syndrome, using Southern blot analysis. This study emphasizes the value of molecular analysis for gene dosage to suggest chromosomal aneuploidy.

  4. 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

  5. [Overexpression of NHE1 suppresses ABCA1 protein expression via increasing calpain activity in RAW264.7 cells].

    PubMed

    Mo, Xiangang; Wang, Lan; Guo, Jing; Hong, Wei; Long, Shiqi; Zhang, Li; Xiang, Ning; Yang, Juan

    2017-01-01

    Objective To investigate the effect of over-expressed Na(+)/H(+) exchanger 1 (NHE1) on the protein expression of adenosine three phosphate binding cassette transporter A1 (ABCA1) in RAW264.7 cells. Methods RAW264.7 cells were infected with the adenoviral vector encoding NHE1-EGFP (AdNHE1). The infected RAW264.7 cells were subjected to Western blot analysis for NHE1-EGFP fusion protein. The subcellular localization of NHE1-EGFP fusion protein was observed by confocal laser scanning microscopy. NHE1 activity was measured by the method of pH recovery in response to an acute acid pulse. Furthermore, Western blotting was performed to determine ABCA1 protein levels and calpain activity in NHE1-overexpressing RAW264.7 cells. The effect of calpain inhibitor N-acetyl-L-leucyl-L-leucyl-L-norleucinal (ALLN) on ABCA1 protein levels in the presence of TO-901317 was examined by Western blotting. Results NHE1-EGFP fusion protein was highly expressed and localized in cytoplasm and cell membrane of RAW264.7 cells infected with AdNHE1. NHE1-EGFP fusion protein reduced ABCA1 protein expression and increased calpain activity. The calpain inhibitor ALLN blocked the decrease of ABCA1 protein expression. Conclusion Overexpressed NHE1 suppresses the expression of ABCA1 protein via increasing the calpain activity in RAW264.7 cells.

  6. Molecular Phenotyping Small (Asian) versus Large (Western) Plaque Psoriasis Shows Common Activation of IL-17 Pathway Genes but Different Regulatory Gene Sets.

    PubMed

    Kim, Jaehwan; Oh, Chil-Hwan; Jeon, Jiehyun; Baek, Yoosang; Ahn, Jaewoo; Kim, Dong Joo; Lee, Hyun-Soo; Correa da Rosa, Joel; Suárez-Fariñas, Mayte; Lowes, Michelle A; Krueger, James G

    2016-01-01

    Psoriasis is present in all racial groups, but in varying frequencies and severity. Considering that small plaque psoriasis is specific to the Asian population and severe psoriasis is more predominant in the Western population, we defined Asian small and intermediate plaque psoriasis as psoriasis subtypes and compared their molecular signatures with the classic subtype of Western large plaque psoriasis. Two different characteristics of psoriatic spreading-vertical growth and radial expansion-were contrasted between subtypes, and genomic data were correlated to histologic and clinical measurements. Compared with Western large plaque psoriasis, Asian small plaque psoriasis revealed limited psoriasis spreading, but IL-17A and IL-17-regulated proinflammatory cytokines were highly expressed. Paradoxically, IL-17A and IL-17-regulated proinflammatory cytokines were lower in Western large plaque psoriasis, whereas T cells and dendritic cells in total psoriatic skin area were exponentially increased. Negative immune regulators, such as CD69 and FAS, were decreased in both Western large plaque psoriasis and psoriasis with accompanying arthritis or obesity, and their expression was correlated with psoriasis severity index. Based on the disease subtype comparisons, we propose that dysregulation of T-cell expansion enabled by downregulation of immune negative regulators is the main mechanism for development of large plaque psoriasis subtypes.

  7. 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

  8. Geology of the Crust and Mantle, Western United States: Geophysical data reveal a thin crust and anomalous upper mantle characteristic of active regions.

    PubMed

    Thompson, G A; Talwani, M

    1964-12-18

    compressional and transverse waves in the upper mantle may be related to this problem. Whatever its origin and composition, an anomalous upper mantle characterizes many regions of present or recent tectonic activity, such as Japan and the Mid-Atlantic Ridge (39). The anomalous mantle of western North America might form a continuous belt to the south, with anomalous mantle beneath the crest of the East Pacific Rise (40). The anomalous upper mantle may thus be an essential part of the heat engine driving the tectonic activity of these regions. The Basin and Range region was broken into blocks and laterally extended during the Cenozoic uplift, so that some blocks lagged behind, or sank. Some of the intricate disruption of the upper crust may be related to shallow Cenozoic volcanism. The relatively large and rigid Sierra Nevada block may have been tilted westward during Basin-Range deformation because of the high density of greenstones on the west side and the lower density of granitic rocks to the east. Man's environment, in the longer view of geologic time, is strongly influenced by mountain-building processes originating in the earth's crust and mantle. In the scale of a few lifetimes, climate, sea level, and the shape of the land are appreciably altered. How this comes about, and whether man can hope to influence the processes, are challenging, unsolved problems. But enough has now been learned about the crust and mantle to suggest precisely what questions must be answered and what critical experiments performed. Note added in proof: Osborne (42) has directed our attention to the possibility that the granitic rocks and also the andesites and dacites were formed by fractional crystallization of basaltic magma under conditions of high oxygen pressure. This possibility in no way conflicts with the geophysical data. In fact, such direct additions to the silicic upper crust from the mantle or lower crust would simplify the perplexing problem of how the crust is replenished in areas

  9. Fibrinogen and Fibronectin Binding Activity and Immunogenic Nature of Choline Binding Protein M

    PubMed Central

    AFSHAR, Davoud; POURMAND, Mohammad Reza; JEDDI-TEHRANI, Mahmood; SABOOR YARAGHI, Ali Akbar; AZARSA, Mohammad; SHOKRI, Fazel

    2016-01-01

    Background: Choline-binding proteins (CBPs) are a group of surface-exposed proteins, which play crucial and physiological roles in Streptococcus pneumoniae. The novel member of CBPs, choline-binding protein M (CbpM) may have binding activity to plasma proteins. This study aimed to clone and express CbpM and demonstrate its interaction with plasma proteins and patients’ sera. Methods: The total length of cbpM gene was cloned in pET21a vector and expressed in BL21 expression host. Verification of recombinant protein was evaluated by Western blot using anti-His tag monoclonal antibody. Binding ability of the recombinant protein to plasma proteins and the interaction with patients’ sera were assessed by Western blot and ELISA methods. Results: The cbpM gene was successfully cloned into pET21a and expressed in BL21 host. Binding activity to fibronectin and fibrinogen and antibody reaction of CbpM to patients’ sera was demonstrated by Western blot and ELISA methods, respectively. Conclusion: CbpM is one of the pneumococcal surface-exposed proteins, which mediates pneumococcal binding to fibronectin and fibrinogen proteins. PMID:28053927

  10. Hepatitis B virus X protein activates human hepatic stellate cells through upregulating TGFβ1.

    PubMed

    Chen, H-Y; Chen, Z-X; Huang, R-F; Lin, N; Wang, X-Z

    2014-10-27

    We investigated the effects of the hepatitis B virus X gene (HBV X) on the activation of human hepatic stellate cells (HSCs) and the possible mechanisms underlying the pathway. Recombinant plasmid pHBV-X-IRES2-EGFP was constructed and transfected into HL-7702 cells using a lipid-mediated method. Transfected cells were screened by G418, which detected stable expression of the X gene by reverse transcription (RT)-PCR and Western blot analysis, and named L02/x. Cells not subjected to G418-selection were analyzed to confirm the transient expression of the X gene and named L02/48x. Subsequently, L02/x and L02/48x, together with non-HBx-expressing cells, were co-cultured with HSCs in a non-contact transwell system. After 36 h of co-culture, the proliferation and migration of HSCs was detected using different cell counting methods. Finally, the mRNA and protein levels of α-SMA, Col I, and TGFβ1 in HSCs were detected by real-time PCR and western blot analysis. RT-PCR and Western blot analysis showed that L02/x and L02/48x cells can express HBV X gene mRNA and protein. Additionally, HSCs co-cultured with L02/x or L02/48x cells showed significantly higher proliferation and migration levels than control groups. Real-time PCR and Western blot analysis showed that the mRNA and protein expressions of α-SMA, Col I, and TGFβ1 in HSCs co-cultured with HBx-expressing liver cells were higher than those in control groups. HBx protein activated HSCs in vitro, leading to increased proliferation and migration of HSCs and upregulation of α-SMA and Col I. The TGFβ1 gene may be involved in this pathway.

  11. Rapid Screening of the Epidermal Growth Factor Receptor Phosphosignaling Pathway via Microplate-Based Dot Blot Assays

    PubMed Central

    Cappione, Amedeo; Smith, Janet; Mabuchi, Masaharu; Nadler, Timothy

    2012-01-01

    Expression profiling on a large scale, as is the case in drug discovery, is often accomplished through use of sophisticated solid-phase protein microarrays or multiplex bead technologies. While offering both high-throughput and high-content analysis, these platforms are often too cost prohibitive or technically challenging for many research settings. Capitalizing on the favorable attributes of the standard ELISA and slot blotting techniques, we developed a modified dot blot assay that provides a simple cost-effective alternative for semiquantitative expression analysis of multiple proteins across multiple samples. Similar in protocol to an ELISA, but based in a membrane bound 96-well microplate, the assay takes advantage of vacuum filtration to expedite the tedious process of washing in between binding steps. We report on the optimization of the assay and demonstrate its use in profiling temporal changes in phosphorylation events in the well-characterized EGF-induced signaling cascade of A431 cells. PMID:22934183

  12. Speciation of mercury in mine waste: case study of abandoned and active gold mine sites at the Bibiani-Anwiaso-Bekwai area of South Western Ghana.

    PubMed

    Nartey, V K; Klake, R K; Doamekpor, L K; Sarpong-Kumankomah, S

    2012-12-01

    Speciation determines toxicity, transport pathways and residence time of a metal in different compartments of the environment. This study investigated the speciation of mercury in soils, derived from sites known for dumping of mine wastes in the Bibiani-Anwiaso-Bekwai district, a gold mining community of the Western Region of Ghana. Soil samples were taken from the surface; depths of 20, 40 and 60 cm from mine waste at both abandoned and active mine sites. Each sample was analysed for total mercury, organic mercury and elemental mercury. After sample treatment, digestion and reduction with stannous chloride (SnCl(2)), total mercury content was determined using the Inductively Coupled Plasma-Optical Emissions Spectrometer (ICP-OES). Organic mercury content was determined employing a differential technique after disposing of elemental mercury by heating. Total mercury content in samples ranged from 0.067 to 0.876 mg/kg for surface soils. The same soil of depths 20, 40 and 60 cm had total mercury from 0.102 to 1.066, 0.037 to 4.037 and 0.191 to 4.998 mg/kg, respectively. For organic mercury, concentrations range from 0.012 to 0.260 mg/kg for surface soil. Soil depths of 20, 40 and 60 cm had organic mercury concentrations from 0.016 to 0.653, 0.041 to 1.093 and 0.101 to 2.546 mg/kg respectively. Elemental mercury concentrations in surface soils, soils at depths of 20, 40 and 60 cm ranged from 0.043 to 0.780; 0.017 to 0.749; 0.014 to 2.944 and 0.009 to 2.452 mg/kg respectively. Among the sites studied, only galamsey tailings (GM) showed a trend of increasing total mercury level with increasing depth. For the other sites, trends were not defined. There has been no defined trend for elemental mercury with depth at any of the sampling sites. Just as with total mercury, it was only GM that showed an increasing trend of organic mercury concentration with depth.

  13. Active tectonics of the western tethyan himalaya above the underthrusting indian plate: The upper sutlej river basin as a pull-apart structure

    NASA Astrophysics Data System (ADS)

    Ni, James; Barazangi, Muawia

    1985-03-01

    Fault-bounded blocks and structural elements were mapped in the eastern Ladakh-Spiti and upper Sutlej River Basin located within the Tethyan Himalaya and to the southwest of the Karakorum fault zone mainly using LANDSAT Multispectral Scanner (MSS) band 5, band 7 (near-infrared) images with detailed analysis of smaller areas by interactive digital processing of false color images, and Returned Beam Vidicon (RBV) imagery in conjunction with available topographical, geological and seismological data. For the first time the Leo Pargil Horst and other nearby fault-bounded blocks located at the northwestern end of the upper Setlej River Basin were clearly revealed on the LANDSAT color composites. Shallow crustal seismicity is systematically related to the NNE-trending and N-trending normal faults of the Leo Pargil and nearby regions. Some of the aftershocks of the Kinnaur earthquake of January 19,1975 ( Ms = 6.8), appear to be associated with movement along the NNE-trending westbound fault of the Leo Pargil Horst and the nearby Kaurik-Chango fault. The main shock, however, is teleseismically located at about 30 km to the northwest of the Kaurik-Chango fault. Fault plane solutions of the main shock and two aftershocks indicate a large component of normal faulting. In map view, the upper Sutlej River Basin has an approximately rhomboidal shape and is located to the southwest of the Karakorum fault system. We suggest that this basin is a pull-apart between the NW-SE oriented, right-lateral, strike-slip Karakorum fault system and the high-angle faults near the southern boundary of the Tethyan Himalaya. The Leo Pargil Horst is the northwestern bounding fault block of this pull-apart. The active tectonic features in this part of the Tethyan Himalaya appear to reflect right-shear within the crust, and this is probably a consequence of oblique underthrusting of the Indian continental plate beneath the western Himalaya and southwestern Tibet during the Neogene and Quaternary

  14. 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...

  15. Female mice lacking active nadph-oxidase enzymes are protected against “western diet”--induced obesity and metabolic syndrome

    Technology Transfer Automated Retrieval System (TEKTRAN)

    NADPH oxidase (Nox) enzymes have been implicated in regulation of adipocyte differentiation and inflammation in a variety of tissues. We examined the effects of feeding AIN-93G or a “Western diet” (WD) (45% fat, 0.5% cholesterol) on development of obesity and “metabolic syndrome” in wild type (WT) m...

  16. 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...

  17. Development of PCR/dot blot assay for specific detection and differentiation of taeniid cestode eggs in canids.

    PubMed

    Armua-Fernandez, Maria Teresa; Nonaka, Nariaki; Sakurai, Tatsuya; Nakamura, Seita; Gottstein, Bruno; Deplazes, Peter; Phiri, Isaac G K; Katakura, Ken; Oku, Yuzaburo

    2011-01-01

    We report the development of a colourimetric PCR/dot blot assay targeting the mitochondrial gene NADH dehydrogenase subunit 1 (nad1) for differential diagnosis of taeniid eggs. Partial sequences of the cestode nad1 gene were aligned and new primers were designed based on conserved regions. Species-specific oligonucleotide probes (S-SONP) for canine taeniid cestodes were then designed manually based on the variable region between the conserved primers. Specifically, S-SONP were designed for the Taenia crassiceps, T. hydatigena, T. multiceps, T. ovis, T. taeniaeformis, Echinococcus granulosus (genotype 1), E. multilocularis and E. vogeli. Each probe showed high specificity as no cross-hybridisation with any amplified nad1 fragment was observed. We evaluated the assay using 49 taeniid egg-positive samples collected from dogs in Zambia. DNA from 5 to 10 eggs was extracted in each sample. Using the PCR/dot blot assay, the probes successfully detected PCR products from T. hydatigena in 42 samples, T. multiceps in 3 samples, and both species (mixed infection) in the remaining 4 samples. The results indicate that the PCR/dot blot assay is a reliable alternative for differential diagnosis of taeniid eggs in faecal samples.

  18. WESTERN ECOLOGY DIVISION - GENERAL INFORMATION SHEET

    EPA Science Inventory

    abstract for flyer - general information The Western Ecology Division (WED), part of EPAs National Health and Environmental Effects Research Laboratory, provides information to EPA offices and regions nationwide to improve understanding of how human activities affect estuarine,...

  19. 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

  20. Detection and chromosomal assignment of SV40-DNA integration in Chinese hamster cell lines by chromosome sorting and dot blot hybridization.

    PubMed

    Hutter, K J; Klefenz, H; Goerttler, K

    1990-01-01

    A combination of cytometric (chromosome sorting), molecular (dot blot hybridization using radio-active and/or biotinylated DNA probes) and cytogenetic (G-banding) evaluation is described which allows the rapid identification of single copy and repetitive viral integrates and their assignment to chromosome groups or even individual chromosomes. In the case of Chinese hamster cell line CO 631 it could be demonstrated that SV40 DNA was solely integrated into a submetacentric marker chromosome. Such a cytometric/molecular/cytogenetic "identogram" may prove to be a useful tool in many areas of cell and tumor biology. Furthermore, amounts of chromosomes sufficient for analysis as well as subsequent cloning experiments can be accumulated.

  1. 17-DMAG Diminishes Hemorrhage-Induced Small Intestine Injury by Elevating Bcl-2 Protein and Inhibiting iNOS Pathway, TNF-alpha Increase, and Caspase-3 Activation

    DTIC Science & Technology

    2011-06-03

    Hemorrhagic shock has been shown to cause systemic inflammation response syndrome (SIRS), multiple organ dysfunction syndrome (MODS), and multiple organ...available to tissues and results in an accumulation of carbon dioxide and metabolic waste , leading to activation of signal transduction pathways and...for each specimen was graded using a six-tiered scale [3]. Western blots. Jejunal tissue was minced in 100 µl Hanks’ balanced salt solution

  2. 5-Androstenediol Promotes Survival of Gamma-Irradiated Human Hematopoietic Progenitors Through Induction of NF-kappa B Activation and G-CSF Expression

    DTIC Science & Technology

    2007-05-01

    and Kroemer G (2006) NF-kappaB constitutes a potential therapeutic target in high-risk myelodysplastic syndrome . Blood 107:1156-1165. Carlsen H...DA, Luger SM and Jordan CT (2001) Nuclear factor-kappaB is constitutively activated in primitive human acute myelogenous leukemia cells. Blood 98...the pro-apoptotic factor p53 after IR, as determined by Western blot. The results indicate that NFkB1 degradation after IR may be responsible for the

  3. Epizootic activity of Murray Valley encephalitis and Kunjin viruses in an aboriginal community in the southeast Kimberley region of Western Australia: results of mosquito fauna and virus isolation studies.

    PubMed

    Broom, Annette K; Lindsay, Michael D A; Wright, Anthony E; Smith, David W; Mackenzie, John S

    2003-09-01

    We undertook annual surveys of flavivirus activity in the community of Billiluna in the southeast Kimberley region of Western Australia between 1989 and 2001 [corrected]. Culex annulirostris was the dominant mosquito species, particularly in years of above average rains and flooding. Murray Valley encephalitis (MVE) virus was isolated in 8 of the 13 years of the study from seven mosquito species, but more than 90% of the isolates were from Cx. annulirostris. The results suggest that MVE virus is epizootic in the region, w ith activity only apparent in years with average or above average rainfall and increased numbers of Cx. annulirostris. High levels of MVE virus activity and associated human cases were detected only once (in 1993) during the survey period. Activity of MVE virus could only be partially correlated with wet season rainfall and flooding, suggesting that a number of other factors must also be considered to accurately predict MVE virus activity at such communities.

  4. Examination of Proteins Bound to Nascent DNA in Mammalian Cells Using BrdU-ChIP-Slot-Western Technique.

    PubMed

    Bhaskara, Srividya

    2016-01-14

    Histone deacetylases 1 and 2 (HDAC1,2) localize to the sites of DNA replication. In the previous study, using a selective inhibitor and a genetic knockdown system, we showed novel functions for HDAC1,2 in replication fork progression and nascent chromatin maintenance in mammalian cells. Additionally, we used a BrdU-ChIP-Slot-Western technique that combines chromatin immunoprecipitation (ChIP) of bromo-deoxyuridine (BrdU)-labeled DNA with slot blot and Western analyses to quantitatively measure proteins or histone modification associated with nascent DNA. Actively dividing cells were treated with HDAC1,2 selective inhibitor or transfected with siRNAs against Hdac1 and Hdac2 and then newly synthesized DNA was labeled with the thymidine analog bromodeoxyuridine (BrdU). The BrdU labeling was done at a time point when there was no significant cell cycle arrest or apoptosis due to the loss of HDAC1,2 functions. Following labeling of cells with BrdU, chromatin immunoprecipitation (ChIP) of histone acetylation marks or the chromatin-remodeler was performed with specific antibodies. BrdU-labeled input DNA and the immunoprecipitated (or ChIPed) DNA was then spotted onto a membrane using the slot blot technique and immobilized using UV. The amount of nascent DNA in each slot was then quantitatively assessed using Western analysis with an anti-BrdU antibody. The effect of loss of HDAC1,2 functions on the levels of newly synthesized DNA-associated histone acetylation marks and chromatin remodeler was then determined by normalizing the BrdU-ChIP signal obtained from the treated samples to the control samples.

  5. 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

  6. 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.

  7. Simultaneous detection of fourteen Italian cystic fibrosis mutations in seven exons by reverse dot-blot analysis.

    PubMed

    Rady, M; D'Alcamo, E; Seia, M; Iapichino, L; Ferrari, M; Russo, S; Romeo, G; Maggio, A

    1995-10-01

    The increasing number of cystic fibrosis (CF) mutations is a great obstacle to the use of DNA procedures in the detection of gene defects. We describe a fast, cheap and non-radioactive procedure, Reverse dot-blot analysis (RDB), for the simultaneous detection of CF mutations in the Italian population. We used this approach to study seven exons of the CF gene for 14 CF gene defects and were able to characterize 222 of 272 CF chromosomes (80%). The cost of the procedure was $25 per sample analysed.

  8. Detection of Rickettsia in Rhipicephalus sanguineus Ticks and Ctenocephalides felis Fleas from Southeastern Tunisia by Reverse Line Blot Assay

    PubMed Central

    Khrouf, Fatma; M'Ghirbi, Youmna; Znazen, Abir; Ben Jemaa, Mounir; Hammami, Adnene

    2014-01-01

    Ticks (n = 663) and fleas (n = 470) collected from domestic animals from southeastern Tunisia were screened for Rickettsia infection using reverse line blot assay. Evidence of spotted fever group Rickettsia was obtained. We detected Rickettsia felis in fleas, Rickettsia massiliae Bar 29 and the Rickettsia conorii Israeli spotted fever strain in ticks, and Rickettsia conorii subsp. conorii and Rickettsia spp. in both arthropods. The sensitivity of the adopted technique allowed the identification of a new association between fleas and R. conorii subsp. conorii species. The presence of these vector-borne Rickettsia infections should be considered when diagnosing this disease in humans in Tunisia. PMID:24226919

  9. RTG resource book for western states and provinces: Final proceedings

    SciTech Connect

    1994-12-31

    The Western Interstate Energy Board held a workshop and liaison activities among western states, provinces, and utilities on the formation of Regional Transmission Groups (RTGs). Purpose of the activities was to examine the policy implications for western states and provinces in the formation of RTGs in the West, the implications for western ratepayers and utilities of the RTG formation and potential impacts of RTGs on the western electricity system. The workshop contributed to fulfilling the transmission access and competition objectives of Title VII of the Energy Policy Act of 1992.

  10. North African dust transport toward the western Mediterranean basin: atmospheric controls on dust source activation and transport pathways during June-July 2013

    NASA Astrophysics Data System (ADS)

    Schepanski, Kerstin; Mallet, Marc; Heinold, Bernd; Ulrich, Max

    2016-11-01

    Dust transported from north African source region toward the Mediterranean basin and Europe is a ubiquitous phenomenon in the Mediterranean region. Winds formed by large-scale pressure gradients foster dust entrainment into the atmosphere over north African dust source regions and advection of dust downwind. The constellation of centers of high and low pressure determines wind speed and direction, and thus the chance for dust emission over northern Africa and transport toward the Mediterranean. We present characteristics of the atmospheric dust life cycle determining dust transport toward the Mediterranean basin with focus on the ChArMEx (Chemistry-Aerosol Mediterranean Experiment) special observation period in June and July 2013 using the atmosphere-dust model COSMO-MUSCAT (COSMO: COnsortium for Small-scale MOdeling; MUSCAT: MUltiScale Chemistry Aerosol Transport Model). Modes of atmospheric circulation are identified from empirical orthogonal function (EOF) analysis of the geopotential height at 850 hPa and compared to EOFs calculated from 1979-2015 ERA-Interim reanalysis. Two different phases are identified from the first EOF, which in total explain 45 % of the variance. They are characterized by the propagation of the subtropical ridge into the Mediterranean basin, the position of the Saharan heat low and the predominant Iberian heat low, and discussed illustrating a dipole pattern for enhanced (reduced) dust emission fluxes, stronger (weaker) meridional dust transport, and consequent increased (decreased) atmospheric dust concentrations and deposition fluxes. In the event of a predominant high-pressure zone over the western and central Mediterranean (positive phase), a hot spot in dust emission flux is evident over the Grand Erg Occidental, and a reduced level of atmospheric dust loading occurs over the western Mediterranean basin. The meridional transport in northward direction is reduced due to prevailing northerly winds. In case of a predominant heat low

  11. Detection of Mycobacterium avium subsp. paratuberculosis by buoyant density centrifugation, sequence capture PCR and dot blot hybridisation.

    PubMed

    Halldórsdóttir, Stefania; Englund, Stina; Nilsen, Sigrun Fredsvold; Olsaker, Ingrid

    2002-07-22

    Detection of Mycobacterium avium subsp. paratuberculosis (M. paratuberculosis) by polymerase chain reaction (PCR) is often hampered by the lack of efficient methods for sample treatment. We report a protocol for analysis of faecal samples based on buoyant density centrifugation in Percoll and IS900 sequence capture PCR combined with a dot blot assay for detection of low-grade infection of M. paratuberculosis. Serial dilutions of M. paratuberculosis genomic DNA and M. paratuberculosis bacteria were used to assess the sensitivity of the method. The final evaluation was performed with spiked faecal samples, which also were analysed by culture. The presence of PCR inhibitory substances in processed faecal samples was evaluated by including a PCR internal control. By using buoyant density centrifugation, sequence capture PCR, and dot blot hybridisation, we achieved a sensitivity of 10(3)CFU (colony forming units)/g of faeces. The detection limit by culture was assessed to 10(2)CFU/g of faeces. We conclude that the described protocol is a fast and sensitive alternative to bacterial culture of faecal samples.

  12. A solid-phase immunoassay of protease-resistant prion protein with filtration blotting involving sodium dodecyl sulfate.

    PubMed

    Kobayashi, Yoshiteru; Kohno, Naoyuki; Wanibe, Shoko; Hirayasu, Kazunari; Uemori, Hitoshi; Tagawa, Yuichi; Yokoyama, Takashi; Shinagawa, Morikazu

    2006-02-15

    The precise diagnosis for bovine spongiform encephalopathy (BSE) is crucial for preventing new transmission to humans. Several testing procedures are reported for determining protease-resistant prion protein in various tissues as a major hallmark of prion diseases such as BSE, scrapie, and Creutzfeldt-Jakob disease. However, contamination of materials from tissues or degradation of the specimens sometimes disturbs the accuracy of the assay. Here, we have developed a novel method for solid-phase immunoassay of the disease-specific conformational isoform, PrP(Sc), using filtration blotting of protein in the presence of sodium dodecyl sulfate (SDS) followed by a filtration-based immunoassay with a single anti-prion protein antibody, together with the improved fractionation procedure involving high concentrations of surfactant/detergent. The SDS/heat treatment renders unfolded PrP(Sc) quantitative retention on a polyvinylidene difluoride filter and allows enhancement of the analyte signal with immunodetection; thus, all of the tested specimens are determined with 100% accuracy. In addition, the immunoassay is completed in approximately 1h, indicating its usefulness not only for the screening of BSE specimens but probably also for the postmortem BSE diagnosis of fallen stock as the antibody recognizes the core part of PrP(Sc). The solid-phase immunoassay method, including the filtration blotting with SDS, would be applicable to determining even more sensitively proteins other than PrP(Sc), especially those having rigid conformations.

  13. 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.

  14. 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.

  15. American ginseng suppresses Western diet-promoted tumorigenesis in model of inflammation-associated colon cancer: role of EGFR

    PubMed Central

    2011-01-01

    Background Western diets increase colon cancer risk. Epidemiological evidence and experimental studies suggest that ginseng can inhibit colon cancer development. In this study we asked if ginseng could inhibit Western diet (20% fat) promoted colonic tumorigenesis and if compound K, a microbial metabolite of ginseng could suppress colon cancer xenograft growth. Methods Mice were initiated with azoxymethane (AOM) and, two weeks later fed a Western diet (WD, 20% fat) alone, or WD supplemented with 250-ppm ginseng. After 1 wk, mice received 2.5% dextran sulfate sodium (DSS) for 5 days and were sacrificed 12 wks after AOM. Tumors were harvested and cell proliferation measured by Ki67 staining and apoptosis by TUNEL assay. Levels of EGF-related signaling molecules and apoptosis regulators were determined by Western blotting. Anti-tumor effects of intraperitoneal compound K were examined using a tumor xenograft model and compound K absorption measured following oral ginseng gavage by UPLC-mass spectrometry. Effects of dietary ginseng on microbial diversity were measured by analysis of bacterial 16S rRNA. Results Ginseng significantly inhibited colonic inflammation and tumorigenesis and concomitantly reduced proliferation and increased apoptosis. The EGFR cascade was up-regulated in colonic tumors and ginseng significantly reduced EGFR and ErbB2 activation and Cox-2 expression. Dietary ginseng altered colonic microbial diversity, and bacterial suppression with metronidazole reduced serum compound K following ginseng gavage. Furthermore, compound K significantly inhibited tumor xenograft growth. Conclusions Ginseng inhibited colonic inflammation and tumorigenesis promoted by Western diet. We speculate that the ginseng metabolite compound K contributes to the chemopreventive effects of this agent in colonic tumorigenesis. PMID:22070864

  16. A Western Pacific Hotspot?

    NASA Astrophysics Data System (ADS)

    MacPherson, C. G.; Hall, R.

    2003-04-01

    The petrology of volcanic rocks from the St. Andrew Strait and helium isotope ratios of backarc lavas from the Manus Basin have been used to propose the existence of an active hotspot beneath the eastern Bismarck Sea (Johnson et al., 1978; Macpherson et al., 1998). The past influence of this hotspot can be assessed by mapping its present location onto a plate tectonic reconstruction of the western Pacific (Macpherson and Hall, 2001). During the Middle Eocene the nascent Izu-Bonin-Mariana (IBM) arc lay above the hotspot. The volume of magma emplaced at the IBM arc at that time substantially exceeds the average magma production rate for mature island arcs. Furthermore, the ultramafic (boninitic) character of much of this magmatism requires elevated temperatures. The geochemistry of contemporaneous magmatism in the backarc resembles ocean island basalts and much of the backarc region experienced significant uplift at that time. All of these features can be explained by the influx of hot, buoyant, chemically distinct mantle beneath the IBM and its hinterland. The lithosphere lying above the hotspot during the later Eocene was subsequently subducted. During the Oligo-Miocene the hotspot was traversed by parts of the Caroline Plate where the Euripik Rise is found. This is an aseismic rise that possesses the geophysical characteristics of thickened oceanic crust formed by excess, basaltic magmatism and is the type of structure that would result from the passage of relatively young oceanic lithosphere over a mantle hotspot. Plate reconstruction for the western Pacific predicts a hotspot trail that is consistent with the Middle Eocene and Oligo-Miocene geology of the IBM and Caroline Plates, respectively (Macpherson and Hall, 2001). Parts of the trail have been disrupted by subsequent sea-floor spreading or lost through subduction but the remaining vestiges are consistent with the action of a thermal anomaly throughout much of the Cenozoic. More speculatively, buoyancy

  17. A Western Pacific Hotspot?

    NASA Astrophysics Data System (ADS)