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Sample records for activity northern blot

  1. 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.

  2. 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.

  3. 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

  4. 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.

  5. 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.

  6. 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

  7. Quantitative Expression Analysis in Brassica napus by Northern Blot Analysis and Reverse Transcription-Quantitative PCR in a Complex Experimental Setting.

    PubMed

    Rumlow, Annekathrin; Keunen, Els; Klein, Jan; Pallmann, Philip; Riemenschneider, Anja; Cuypers, Ann; Papenbrock, Jutta

    Analysis of gene expression is one of the major ways to better understand plant reactions to changes in environmental conditions. The comparison of many different factors influencing plant growth challenges the gene expression analysis for specific gene-targeted experiments, especially with regard to the choice of suitable reference genes. The aim of this study is to compare expression results obtained by Northern blot, semi-quantitative PCR and RT-qPCR, and to identify a reliable set of reference genes for oilseed rape (Brassica napus L.) suitable for comparing gene expression under complex experimental conditions. We investigated the influence of several factors such as sulfur deficiency, different time points during the day, varying light conditions, and their interaction on gene expression in oilseed rape plants. The expression of selected reference genes was indeed influenced under these conditions in different ways. Therefore, a recently developed algorithm, called GrayNorm, was applied to validate a set of reference genes for normalizing results obtained by Northern blot analysis. After careful comparison of the three methods mentioned above, Northern blot analysis seems to be a reliable and cost-effective alternative for gene expression analysis under a complex growth regime. For using this method in a quantitative way a number of references was validated revealing that for our experiment a set of three references provides an appropriate normalization. Semi-quantitative PCR was prone to many handling errors and difficult to control while RT-qPCR was very sensitive to expression fluctuations of the reference genes.

  8. Quantitative Expression Analysis in Brassica napus by Northern Blot Analysis and Reverse Transcription-Quantitative PCR in a Complex Experimental Setting

    PubMed Central

    Rumlow, Annekathrin; Keunen, Els; Klein, Jan; Pallmann, Philip; Riemenschneider, Anja; Cuypers, Ann

    2016-01-01

    Analysis of gene expression is one of the major ways to better understand plant reactions to changes in environmental conditions. The comparison of many different factors influencing plant growth challenges the gene expression analysis for specific gene-targeted experiments, especially with regard to the choice of suitable reference genes. The aim of this study is to compare expression results obtained by Northern blot, semi-quantitative PCR and RT-qPCR, and to identify a reliable set of reference genes for oilseed rape (Brassica napus L.) suitable for comparing gene expression under complex experimental conditions. We investigated the influence of several factors such as sulfur deficiency, different time points during the day, varying light conditions, and their interaction on gene expression in oilseed rape plants. The expression of selected reference genes was indeed influenced under these conditions in different ways. Therefore, a recently developed algorithm, called GrayNorm, was applied to validate a set of reference genes for normalizing results obtained by Northern blot analysis. After careful comparison of the three methods mentioned above, Northern blot analysis seems to be a reliable and cost-effective alternative for gene expression analysis under a complex growth regime. For using this method in a quantitative way a number of references was validated revealing that for our experiment a set of three references provides an appropriate normalization. Semi-quantitative PCR was prone to many handling errors and difficult to control while RT-qPCR was very sensitive to expression fluctuations of the reference genes. PMID:27685087

  9. 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.

  10. Analysis of solvent control and 1-nitrosopyrene-induced chinese hamster ovary cell mutants by southern and northern blots and the polymerase chain reaction

    SciTech Connect

    Newton, R.K.; Mittelstaedt, R.A.; Heflich, R.H. )

    1992-01-01

    1-Nitrosopyrene, a metabolite of the tumorigenic environmental pollutant 1-nitropyrene, is a potent mutagen at the hprt locus in Chinese hamster ovary (CHO) cells. A single DNA adduct, N-(deoxyguanosin-8-yl)-l-aminopyrene, is produced in CHO cells treated with 1-nitrosopyrene, and this adduct is found in rats and mice exposed to 1-nitropyrene. In this study, the structure of the hprt gene and the structure and amount of hprt mRNA were analyzed in 43 CHO cell mutants (16 isolated from solvent control cultures and 27 isolated from 1-nitrosopyrene-treated cultures). PstI- and BamHI-digested DNA from the mutants were subjected to Southern blot analysis using a hamster hprt cDNA probe. None of the 1-nitrosopyrene-induced mutants and only one of the control mutants displayed hybridization patterns that were different from the parent CHO cells. Northern blot analysis revealed that two control mutants had truncated hprt mRNAs, while 56% of the control mutants and 78% of the induced mutants had reduced levels of hprt mRNA. Using polymerase chain reaction amplification of cDNA synthesized from RNA, the hprt protein-coding region could be amplified from 23 of the 1-nitrosopyrene-induced mutants and 11 of the control mutants. The amplification products from 3 of the control mutants and 5 of the induced mutants were smaller than that found with RNA from parental CHO cells. These results indicate that the mutagenic DNA damage produced by 1-nitrosopyrene in CHO cells does not cause major structural alterations in the hprt gene and suggest that 1-nitrosopyrene acts as a point mutagen. A large number of both control and 1-nitrosopyrene-induced mutants exhibited a marked reduction in hprt mRNA concentration or possessed truncated mRNA hprt protein coding sequences. These alterations may contribute to the 6-thioguanine-resistant phenotype.

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

    PubMed Central

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

    2011-01-01

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

  12. Whole genomic sequencing of RT98 mitochondria derived from Oryza rufipogon and northern blot analysis to uncover a cytoplasmic male sterility-associated gene.

    PubMed

    Igarashi, Keisuke; Kazama, Tomohiko; Motomura, Keiji; Toriyama, Kinya

    2013-02-01

    Cytoplasmic male sterility (CMS) is a maternally inherited trait resulting in the failure to produce functional pollen and is often observed when an alien cytoplasm is transferred into a cultivated species. An RT98A CMS line and an RT98C fertility restorer line were obtained by successive backcrossing between Oryza rufipogon W1109 and Oryza sativa cultivar Taichung 65. To uncover the CMS-associated mitochondrial genes, we determined the complete sequence of the RT98-CMS mitochondrial genome using next-generation pyrosequencing, and searched new open reading frames (orfs) absent in a reported mitochondrial genome of O. sativa Nipponbare. Then, six candidates were selected for the CMS-associated genes based on the criteria in which they were chimeric in structure or encoded a peptide with transmembrane domains. One of the candidates, orf113, showed different transcript sizes between RT98A and RT98C on Northern blot analysis. The orf113 gene was shown to be co-transcribed with atp4 and cox3 encoding ATP synthase F0 subunit 4 and Cyt c oxidase subunit 3, respectively, and their transcripts were distinctly processed in the presence of a fertility restorer gene. Our results indicate that orf113 is a CMS-associated gene of RT98-CMS.

  13. 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).

  14. Illuminating Northern California's Active Faults

    NASA Astrophysics Data System (ADS)

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

    2009-02-01

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

  15. 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...

  16. 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)

  17. 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.

  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. Illuminating Northern California’s Active Faults

    USGS Publications Warehouse

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

    2009-01-01

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

  20. Hydrothermal Activity in the Northern Guaymas Basin

    NASA Astrophysics Data System (ADS)

    Berndt, C.; Hensen, C.; Mortera-Gutierrez, C. A.; Sarkar, S.; Geilert, S.; Schmidt, M.; Liebetrau, V.; Kipfer, R.; Scholz, F.; Doll, M.; Muff, S.; Karstens, J.; Böttner, C.; Chi, W. C.; Moser, M.; Behrendt, R.; Fiskal, A.; Evans, T.; Planke, S.; Lizarralde, D.; Lever, M. A.

    2015-12-01

    Rift-related magmatism in the Guaymas Basin, Gulf of California induces hydrothermal activity within the basin sediments. Mobilized fluids migrate to the seafloor where they are emitted into the water column changing ocean chemistry and fuelling chemosynthetic ecosystems. New seismic and geochemical data from the northern rift arm of the Guaymas Basin document the variety of fluid expulsion phenomena from large-scale subsurface sediment mobilization related to contact metamorphosis to focused small-scale structures. The geochemical composition of emitted fluids depends largely on the age of the fluid escape structures with respect to the underlying intrusions. Whereas, old structures are dominated by methane emission, young vent sites are characterized by hot fluids that carry a wide range of minerals in solution. The overall high geothermal gradient within the basin (mainly between 160 and 260 °C/km) leads to a thin gas hydrate stability zone. Thus, deep hydrothermal fluid advection affects the gas hydrate system and makes it more dynamic than in colder sedimentary basins.

  1. BLOT Ver. 1.65

    SciTech Connect

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

    2009-03-24

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

  2. Hydrothermal and tectonic activity in northern Yellowstone Lake, Wyoming

    USGS Publications Warehouse

    Johnson, S.Y.; Stephenson, W.J.; Morgan, L.A.; Shanks, Wayne C.; Pierce, K.L.

    2003-01-01

    Yellowstone National Park is the site of one of the world's largest calderas. The abundance of geothermal and tectonic activity in and around the caldera, including historic uplift and subsidence, makes it necessary to understand active geologic processes and their associated hazards. To that end, we here use an extensive grid of high-resolution seismic reflection profiles (???450 km) to document hydrothermal and tectonic features and deposits in northern Yellowstone Lake. Sublacustrine geothermal features in northern Yellowstone Lake include two of the largest known hydrothermal explosion craters, Mary Bay and Elliott's. Mary Bay explosion breccia is distributed uniformly around the crater, whereas Elliott's crater breccia has an asymmetric distribution and forms a distinctive, ???2-km-long, hummocky lobe on the lake floor. Hydrothermal vents and low-relief domes are abundant on the lake floor; their greatest abundance is in and near explosion craters and along linear fissures. Domed areas on the lake floor that are relatively unbreached (by vents) are considered the most likely sites of future large hydrothermal explosions. Four submerged shoreline terraces along the margins of northern Yellowstone Lake add to the Holocene record or postglacial lake-level fluctuations attributed to "heavy breathing" of the Yellowstone magma reservoir and associated geothermal system. The Lake Hotel fault cuts through northwestern Yellowstone Lake and represents part of a 25-km-long distributed extensional deformation zone. Three postglacial ruptures indicate a slip rate of ???0.27 to 0.34 mm/yr. The largest (3.0 m slip) and most recent event occurred in the past ???2100 yr. Although high heat flow in the crust limits the rupture area of this fault zone, future earthquakes of magnitude ???5.3 to 6.5 are possible. Earthquakes and hydrothermal explosions have probably triggered landslides, common features around the lake margins. Few high-resolution seismic reflection surveys have

  3. 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,…

  4. 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.

  5. Project DAFNE - Drilling Active Faults in Northern Europe

    NASA Astrophysics Data System (ADS)

    Kukkonen, I. T.; Ask, M. S. V.; Olesen, O.

    2012-04-01

    We are currently developing a new ICDP project 'Drillling Active Faults in Northern Europe' (DAFNE) which aims at investigating, via scientific drilling, the tectonic and structural characteristics of postglacial (PG) faults in northern Fennoscandia, including their hydrogeology and associated deep biosphere [1, 2]. During the last stages of the Weichselian glaciation (ca. 9,000 - 15,000 years B.P.), reduced ice load and glacially affected stress field resulted in active faulting in Fennoscandia with fault scarps up to 160 km long and 30 m high. These postglacial (PG) faults are usually SE dipping, SW-NE oriented thrusts, and represent reactivated, pre-existing crustal discontinuities. Postglacial faulting indicates that the glacio-isostatic compensation is not only a gradual viscoelastic phenomenon, but includes also unexpected violent earthquakes, suggestively larger than other known earthquakes in stable continental regions. The research is anticipated to advance science in neotectonics, hydrogeology and deep biosphere studies, and provide important information for nuclear waste and CO2 disposal, petroleum exploration on the Norwegian continental shelf and studies of mineral resources in PG fault areas. We expect that multidisciplinary research applying shallow and deep drilling of postglacial faults would provide significant scientific results through generating new data and models, namely: (1) Understanding PG fault genesis and controls of their locations; (2) Deep structure and depth extent of PG faults; (3) Textural, mineralogical and physical alteration of rocks in the PG faults; (4) State of stress and estimates of paleostress of PG faults; (5) Hydrogeology, hydrochemistry and hydraulic properties of PG faults; (6) Dating of tectonic reactivation(s) and temporal evolution of tectonic systems hosting PG faults; (7) Existence/non-existence of deep biosphere in PG faults; (8) Data useful for planning radioactive waste disposal in crystalline bedrock; (9) Data

  6. Electromagnetic study of the active continental margin in northern Chile

    NASA Astrophysics Data System (ADS)

    Echternacht, Friedrich; Tauber, Sebastian; Eisel, Markus; Brasse, Heinrich; Schwarz, Gerhard; Haak, Volker

    1997-06-01

    Magnetotelluric and geomagnetic deep sounding measurements were carried out in the magmatic arc and forearc regions of northern Chile between 19.5° and 22°S to study the electrical conductivity structures of this active continental margin. The instruments used covered a very broad period range from 10 -4 s to approx. 2 × 10 4 s and thus enabled a resolution of deep as well as shallow structures. In this paper we focus on the interpretation of data from an east-west profile crossing Chile from the Pacific coast to the Western Cordillera at 20.5°S. A decomposition of the impedance tensors using the Groom-Bailey decomposition scheme shows that a two-dimensional interpretation is possible. The resulting regional strike direction is N9°W. Two-dimensional models were calculated in this coordinate frame and include the significant bathymetry of the trench as well as the topography of the Andes. The final model shows a generally high resistivity in the forearc and a very good conductor below the Precordillera. Unlike earlier models from areas further south, a good conductor is not observed below the magmatic arc itself. This correlates with the so-called Pica gap in the volcanic chain and a higher age of volcanic activity compared with adjacent areas.

  7. 2000 years of human activity in Tuchola Pinewoods (northern Poland)

    NASA Astrophysics Data System (ADS)

    Obremska, Milena; Ott, Florian; Słowiński, Michał; Lutyńska, Monika; Błaszkiewicz, Mirosław; Brauer, Achim

    2014-05-01

    During the last two millennia human activity and their settlements together with varying climate conditions strongly influenced landscape scale changes. Especially within palaeoecological records these environmental responses are well expressed. However, a robust age control is needed for the evaluation and interpretation of biotic proxies.We present a record from the annually laminated (varved) sediments of Lake Czechowskie, located in northern Poland. The investigated record covers the past 2000 years and demonstrates the continuous vegetation history and human activity in the Northern part of the Tuchola Pinewoods. The chronology was established by varve counting and confirmed by AMS 14C dating, 137Cs activity measurement and a tephra layer (Askja 1875). We used high-resolution biotic (pollen, green algae and diatom analysis) sedimentological (varve and sublayer thickness variations) and geochemical (µ-XRF data) proxies to reconstruct the environmental changes within a time of increasing human activity and fluctuating climatic conditions. Based on different spatial sampling and measuring increments the temporal resolution varies between subseasonal (µ-XRF), annual (varves) up to five-varveresolution (biotic proxies) making it possible to trace even short lasting local and regional changes. Our results display visible human pressure in this area between 50- 350 yr. AD (Roman Period) exerted by tribes related to the Wielbark Culture. The development of persisting settlements and agriculture took place at expense of surrounding hornbeam forests. An intensification of lake productivity (expressed as an increase of varve thickness) started after 250 AD. If this lake ecosystem response relates to an intensified agriculture (and a possible transport of nutrients from neighboring rural lands) or to a climate shift will be further discussed. The rapid decline of human indicators about 350 years AD at the transition to the migration period might be related to cooler

  8. Streamlined Strategies to Better Visualize Southern Blotting

    ERIC Educational Resources Information Center

    Dean, Derek M.

    2012-01-01

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

  9. Blot 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

  10. 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.

  11. 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.

  12. Nocturnal activity patterns of northern myotis (Myotis septentrionalis) during the maternity season in West Virginia (USA)

    USGS Publications Warehouse

    Johnson, J.B.; Edwards, J.W.; Ford, W.M.

    2011-01-01

    Nocturnal activity patterns of northern myotis (Myotis septentrionalis) at diurnal roost trees remain largely uninvestigated. For example, the influence of reproductive status, weather, and roost tree and surrounding habitat characteristics on timing of emergence, intra-night activity, and entrance at their roost trees is poorly known. We examined nocturnal activity patterns of northern myotis maternity colonies during pregnancy and lactation at diurnal roost trees situated in areas that were and were not subjected to recent prescribed fires at the Fernow Experimental Forest, West Virginia from 2007 to 2009. According to exit counts and acoustic data, northern myotis colony sizes were similar between reproductive periods and roost tree settings. However, intra-night activity patterns differed slightly between reproductive periods and roost trees in burned and non-burned areas. Weather variables poorly explained variation in activity patterns during pregnancy, but precipitation and temperature were negatively associated with activity patterns during lactation. ?? Museum and Institute of Zoology PAS.

  13. North Atlantic sea surface temperature, solar activity and the climate of Northern Fennoscandia

    NASA Astrophysics Data System (ADS)

    Ogurtsov, M.; Lindholm, M.; Jalkanen, R.; Veretenenko, S. V.

    2017-02-01

    Seven proxies of summer temperature in Northern Fennoscandia, sea surface temperature in the North Atlantic and solar activity were analyzed over AD 1567-1986. A stable and significant positive correlation between summer temperatures in Northern Fennoscandia and sea surface temperature in the North Atlantic is shown to exist during the entire time interval. In addition, a significant correlation between solar activity and (a) summer temperature in Northern Fennoscandia as well as (b) surface temperature in the North Atlantic was found during AD 1715-1986. Throughout 1567-1715 correlation is less significant and has an opposite sign. Thus we show that the variation of sea surface temperature in the North Atlantic could be a physical agent, which transferred solar influence on Northern Fennoscandian temperature at least during AD 1715-1986.

  14. 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.

  15. The earliest record of human activity in northern Europe

    NASA Astrophysics Data System (ADS)

    Parfitt, Simon A.; Barendregt, René W.; Breda, Marzia; Candy, Ian; Collins, Matthew J.; Coope, G. Russell; Durbidge, Paul; Field, Mike H.; Lee, Jonathan R.; Lister, Adrian M.; Mutch, Robert; Penkman, Kirsty E. H.; Preece, Richard C.; Rose, James; Stringer, Christopher B.; Symmons, Robert; Whittaker, John E.; Wymer, John J.; Stuart, Anthony J.

    2005-12-01

    The colonization of Eurasia by early humans is a key event after their spread out of Africa, but the nature, timing and ecological context of the earliest human occupation of northwest Europe is uncertain and has been the subject of intense debate. The southern Caucasus was occupied about 1.8 million years (Myr) ago, whereas human remains from Atapuerca-TD6, Spain (more than 780kyr ago) and Ceprano, Italy (about 800kyr ago) show that early Homo had dispersed to the Mediterranean hinterland before the Brunhes-Matuyama magnetic polarity reversal (780kyr ago). Until now, the earliest uncontested artefacts from northern Europe were much younger, suggesting that humans were unable to colonize northern latitudes until about 500kyr ago. Here we report flint artefacts from the Cromer Forest-bed Formation at Pakefield (52°N), Suffolk, UK, from an interglacial sequence yielding a diverse range of plant and animal fossils. Event and lithostratigraphy, palaeomagnetism, amino acid geochronology and biostratigraphy indicate that the artefacts date to the early part of the Brunhes Chron (about 700kyr ago) and thus represent the earliest unequivocal evidence for human presence north of the Alps.

  16. 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

  17. 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

  18. 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

  19. Activity of the northern and southern hemispheres as a basis of the solar cycle manifestation

    NASA Astrophysics Data System (ADS)

    Ryabov, M. I.

    2015-12-01

    The dynamics of the evolution of solar cycles is considered a result of the activity manifestation in the northern and southern hemispheres. A study was performed based on separate datasets for the northern and southern hemispheres that contain the monthly and daily averages of the areas of sunspot groups for the period covering activity cycles from 12 to 24 (1874-2013) and the daily values of the Wolf number in the northern and southern hemispheres during cycles 23-24 (1992-2013).To obtain a pattern of development of the "northern" and "southern" solar cycles in detail, a special technique for the extended application of the wavelet analysis has been developed. It allows different the wave processes forming a solar cycle to be distinguished, together with the time of their existence. The application of bandpass Fourier filtering to the obtained data shows that the length of "11-year" cycles by the index S p varies from 10.2 to 11.5 years in the northern hemisphere and from 9.7 to 13.2 years in the southern. The 19th "northern" and 18th "southern" cycles turned out to be maximal. The formation of each of the cycles by all activity indices is determined by the joint effect of long-period processes lasting from 3 to 7 years and short-period processes lasting less than 2 years. When moving from one cycle to another, the long-period processes demonstrate mergings, separations, modulation, and periodic decays. The abnormal activity that appears during the growth, maximum, or decay phase of a cycle is formed at the expense of the simultaneous strengthening of short-period processes, the lengths and "period spectra" of which noticeably differ in the northern and southern hemispheres.

  20. Atlantic SSTs control regime shifts in forest fire activity of Northern Scandinavia

    PubMed Central

    Drobyshev, Igor; Bergeron, Yves; Vernal, Anne de; Moberg, Anders; Ali, Adam A.; Niklasson, Mats

    2016-01-01

    Understanding the drivers of the boreal forest fire activity is challenging due to the complexity of the interactions driving fire regimes. We analyzed drivers of forest fire activity in Northern Scandinavia (above 60 N) by combining modern and proxy data over the Holocene. The results suggest that the cold climate in northern Scandinavia was generally characterized by dry conditions favourable to periods of regionally increased fire activity. We propose that the cold conditions over the northern North Atlantic, associated with low SSTs, expansion of sea ice cover, and the southward shift in the position of the subpolar gyre, redirect southward the precipitation over Scandinavia, associated with the westerlies. This dynamics strengthens high pressure systems over Scandinavia and results in increased regional fire activity. Our study reveals a previously undocumented teleconnection between large scale climate and ocean dynamics over the North Atlantic and regional boreal forest fire activity in Northern Scandinavia. Consistency of the pattern observed annually through millennium scales suggests that a strong link between Atlantic SST and fire activity on multiple temporal scales over the entire Holocene is relevant for understanding future fire activity across the European boreal zone. PMID:26940995

  1. Atlantic SSTs control regime shifts in forest fire activity of Northern Scandinavia.

    PubMed

    Drobyshev, Igor; Bergeron, Yves; Vernal, Anne de; Moberg, Anders; Ali, Adam A; Niklasson, Mats

    2016-03-04

    Understanding the drivers of the boreal forest fire activity is challenging due to the complexity of the interactions driving fire regimes. We analyzed drivers of forest fire activity in Northern Scandinavia (above 60 N) by combining modern and proxy data over the Holocene. The results suggest that the cold climate in northern Scandinavia was generally characterized by dry conditions favourable to periods of regionally increased fire activity. We propose that the cold conditions over the northern North Atlantic, associated with low SSTs, expansion of sea ice cover, and the southward shift in the position of the subpolar gyre, redirect southward the precipitation over Scandinavia, associated with the westerlies. This dynamics strengthens high pressure systems over Scandinavia and results in increased regional fire activity. Our study reveals a previously undocumented teleconnection between large scale climate and ocean dynamics over the North Atlantic and regional boreal forest fire activity in Northern Scandinavia. Consistency of the pattern observed annually through millennium scales suggests that a strong link between Atlantic SST and fire activity on multiple temporal scales over the entire Holocene is relevant for understanding future fire activity across the European boreal zone.

  2. Development of 23 Cycles Activity in Northern and Southern Hemispheres of the Sun

    NASA Astrophysics Data System (ADS)

    Ryabov, M. I.; Sukharev, A. L.; Lukashuk, S. A.

    On an example of the 23rd cycle of solar activity the basic properties of its development in northern and southern hemispheres were researched (daily values of Wolf Sunspot Number - W, the daily values of Sun spots areas - Sp and daily values of flare index - FI.) Effects of application of full-scale Wavelet analysis for studying the temporary structure of formation of a solar cycle show the difference in northern and southern hemispheres. The "leading" periods (in northern hemisphere - 340 days, and in southern hemisphere - 709 days) differ. Thus, the activity period in two years is predominate in southern hemisphere. The "leading" period for the flare index for northern hemisphere is 555 days, and for southern hemisphere is 709 days. In general the basic periods W in northern hemisphere are in the range of 37-555 days, and in southern - 78-906 days, SpN (61-906 days), SpS (61-1477 days), FIN (37-555 days), FIS (23-709 days), FIS - (14709 days) depending on the phase of solar cycle.

  3. Active deformation in the northern Sierra de Valle Fértil, Sierras Pampeanas, Argentina

    NASA Astrophysics Data System (ADS)

    Ortiz, Gustavo; Alvarado, Patricia; Fosdick, Julie C.; Perucca, Laura; Saez, Mauro; Venerdini, Agostina

    2015-12-01

    The Western Sierras Pampeanas region in the San Juan Province is characterized by thick-skinned deformation with approximately N-S trending ranges of average heights of 2500 m and a high frequency occurrence of seismic activity. Its location to the east of the mainly thin-skinned tectonics of the Argentine Precordillera fold-and-thrust belt suggests that at 30°S, deformation is concentrated in a narrow zone involving these two morphostructural units. In this paper, we present new apatite (U-Th)/He results (AHe) across the northern part of the Sierra de Valle Fértil (around 30°S) and analyze them in a framework of thermochronologic available datasets. We found Pliocene AHe results for Carboniferous and Triassic strata in the northern Sierra de Valle Fértil consistent with the hypothesis of recent cooling and inferred erosional denudation concentrated along the northern end of this mountain range. Our analysis shows that this northern region may have evolved under different conditions than the central part of the Sierra de Valle Fértil. Previous studies have observed AHe ages consistent with Permian through Cretaceous cooling, indicating the middle part of the Sierra de Valle Fértil remained near surface before the Pampean slab subduction flattening process. Those studies also obtained ˜5 My cooling ages in the southern part of the Sierra de Valle Fértil, which are similar to our results in the northern end of the range. Taken together, these results suggest a pattern of young deformation in the northern and southern low elevation ends of the Sierra de Valle Fértil consistent with regions of high seismic activity, and Quaternary active faulting along the western-bounding thrust fault of the Sierra de Valle Fértil.

  4. Regulation of Cytokinin Oxidase Activity in Callus Tissues of Phaseolus vulgaris L. cv Great Northern 1

    PubMed Central

    Chatfield, J. Mark; Armstrong, Donald J.

    1986-01-01

    The regulation of cytokinin oxidase activity in callus tissues of Phaseolus vulgaris L. cv Great Northern has been examined using an assay based on the oxidation of N6-(Δ2-isopentenyl)adenine-8-14C (i6 Ade-8-14C) to adenine. Solutions of exogenous cytokinins applied directly to the surface of the callus tissues induced relatively rapid increases in cytokinin oxidase activity. The increase in activity was detectable after 1 hour and continued for about 8 hours, reaching values two- to three-fold higher than the controls. The cytokinin-induced increase in cytokinin oxidase activity was inhibited in tissues pretreated with cordycepin or cycloheximide, suggesting that RNA and protein synthesis may be required for the response. Rifampicin and chloramphenicol, at concentrations that inhibited the growth of Great Northern callus tissues, were ineffective in inhibiting the increase in activity. All cytokinin-active compounds tested, including both substrates and nonsubstrates of cytokinin oxidase, were effective in inducing elevated levels of the enzyme in Great Northern callus tissue. The cytokinin-active urea derivative, Thidiazuron, was as effective as any adenine derivative in inducing this response. The addition of Thidiazuron to the reaction volumes used to assay cytokinin oxidase activity resulted in a marked inhibition of the degradation of the labeled i6 Ade-8-14C substrate. On the basis of this result, it is possible that Thidiazuron may serve as a substrate for cytokinin oxidase, but other mechanisms of inhibition have not yet been excluded. PMID:16664650

  5. 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.

  6. 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.

  7. Bacterial and Fungal Activities of Northern Peatland Ecosystems

    NASA Astrophysics Data System (ADS)

    Winsborough, C.; Basiliko, N.

    2009-05-01

    High latitude peatlands play a unique role in global climate through the long-term net sequestration of atmospheric carbon dioxide in organic soils. Fungi and bacteria dominate microfloral communities in soils and typically are responsible for the majority of direct organic matter decomposition and mineralization, yet each of these groups of microorganisms, with physiological and metabolic differences, potentially plays unique roles in nutrient and carbon cycling in soils. The ability to characterize fungal v bacterial decomposition in peatlands is therefore exceptionally important to understand and predict peatland carbon dynamics, particularly under changing environmental conditions. Here, we demonstrate for the first time, the potential of applying the glucose induced selective inhibition technique, previously used in partitioning bacterial and fungal respiration in forest and agricultural systems, to peatland soils. Using 3 ecologically and hydrologically diverse and spatially dispersed peatlands ranging from a bog to a rich fen, we demonstrated a slight bacterial dominance in a bog and a poor fen both with acidic and primarily Sphagnum peat and a strong bacterial dominance in a near pH neutral, wetter rich fen with sedge peat. This is interesting, as it was expected that microbial respiration in the surface peat profile would be dominated by fungi owing to the acidic and better drained conditions, as is the case with upland riparian forest soils. Furthermore, the maximum non-target inhibition was only 20%, indicating that the SI approach in organic wetland soils works as well as, or better than in, many upland agricultural and forest soils. As the overall importance of fungal and bacterial activities in peatland carbon cycling is still not fully understood, further applications of this technique can develop our understanding of microbial activity in peatland soils.

  8. Observations of the northern seasonal polar cap on Mars: I. Spring sublimation activity and processes

    USGS Publications Warehouse

    Hansen, C.J.; Byrne, S.; Portyankina, G.; Bourke, M.; Dundas, C.; McEwen, A.; Mellon, M.; Pommerol, A.; Thomas, N.

    2013-01-01

    Spring sublimation of the seasonal CO2 northern polar cap is a dynamic process in the current Mars climate. Phenomena include dark fans of dune material propelled out onto the seasonal ice layer, polygonal cracks in the seasonal ice, sand flow down slipfaces, and outbreaks of gas and sand around the dune margins. These phenomena are concentrated on the north polar erg that encircles the northern residual polar cap. The Mars Reconnaissance Orbiter has been in orbit for three Mars years, allowing us to observe three northern spring seasons. Activity is consistent with and well described by the Kieffer model of basal sublimation of the seasonal layer of ice applied originally in the southern hemisphere. Three typical weak spots have been identified on the dunes for escape of gas sublimed from the bottom of the seasonal ice layer: the crest of the dune, the interface of the dune with the interdune substrate, and through polygonal cracks in the ice. Pressurized gas flows through these vents and carries out material entrained from the dune. Furrows in the dunes channel gas to outbreak points and may be the northern equivalent of southern radially-organized channels (“araneiform” terrain), albeit not permanent. Properties of the seasonal CO2 ice layer are derived from timing of seasonal events such as when final sublimation occurs. Modification of dune morphology shows that landscape evolution is occurring on Mars today, driven by seasonal activity associated with sublimation of the seasonal CO2 polar cap.

  9. Antimicrobial activity of three baccharis species used in the traditional medicine of Northern Chile.

    PubMed

    Morales, Glauco; Paredes, Adrián; Sierra, Patricia; Loyola, Luis A

    2008-04-06

    The antimicrobial activities of aqueous ethanol and chloroform extracts of three Baccharis species currently used in Northern Chile folk medicine for the treatment of several infectious and inflammatory disorders were tested against Gram-positive and negative bacteria and fungal spp. using the agar-disc diffusion assay. The results indicated that the activity was more pronounced against Gram-positive than against Gram-negative bacteria and yeast. No significant differences on the antibacterial activity were observed in the aqueous ethanol versus chloroform extracts. None of the plant extracts evaluated exhibited any activity against ten fungi tested.

  10. Nocturnal activity of a "diurnal" species, the northern chamois, in a predator-free Alpine area.

    PubMed

    Carnevali, Lucilla; Lovari, Sandro; Monaco, Andrea; Mori, Emiliano

    2016-05-01

    The reduction of predation risk is widely considered a major factor affecting the nocturnal activity of mammals. Furthermore, on precipitous mountain terrain, moving in very poor light conditions should be avoided by animals with no special eyesight adaptation to darkness. The Northern chamois Rupicapra rupicapra has been for long considered as a diurnal species, with occasional nocturnal movements. For the first time, we have quantified the nocturnal activity of 21 radiotagged female chamois from the Italian Eastern Alps (Paneveggio-Pale di San Martino Natural Park), continuously monitored for two years from sunset to sunrise, with 24h tracking sessions carried out for six months. Large predators were not present in the study site. Despite their mainly diurnal activity pattern, peaks of nocturnal movements were detected throughout the year. The least proportion of active night fixes occurred in January and in July, while the most were in April and in October. The greater nocturnal activity in the warm months compared to cold periods, was probably due to frozen snow cover reducing nocturnal movements. Movements were mainly concentrated in bright moonlight nights, possibly because of the absence of large predators, but more likely because of increased visibility. Changes in activity levels throughout the year may also reflect changes in energy requirements of Northern chamois.

  11. Fault zone structure and inferences on past activities of the active Shanchiao Fault in the Taipei metropolis, northern Taiwan

    NASA Astrophysics Data System (ADS)

    Chen, C.; Lee, J.; Chan, Y.; Lu, C.

    2010-12-01

    The Taipei Metropolis, home to around 10 million people, is subject to seismic hazard originated from not only distant faults or sources scattered throughout the Taiwan region, but also active fault lain directly underneath. Northern Taiwan including the Taipei region is currently affected by post-orogenic (Penglai arc-continent collision) processes related to backarc extension of the Ryukyu subduction system. The Shanchiao Fault, an active normal fault outcropping along the western boundary of the Taipei Basin and dipping to the east, is investigated here for its subsurface structure and activities. Boreholes records in the central portion of the fault were analyzed to document the stacking of post- Last Glacial Maximum growth sediments, and a tulip flower structure is illuminated with averaged vertical slip rate of about 3 mm/yr. Similar fault zone architecture and post-LGM tectonic subsidence rate is also found in the northern portion of the fault. A correlation between geomorphology and structural geology in the Shanchiao Fault zone demonstrates an array of subtle geomorphic scarps corresponds to the branch fault while the surface trace of the main fault seems to be completely erased by erosion and sedimentation. Such constraints and knowledge are crucial in earthquake hazard evaluation and mitigation in the Taipei Metropolis, and in understanding the kinematics of transtensional tectonics in northern Taiwan. Schematic 3D diagram of the fault zone in the central portion of the Shanchiao Fault, displaying regional subsurface geology and its relation to topographic features.

  12. Effects of human subsistence activities on forest birds in northern Kenya.

    PubMed

    Borghesio, Luca

    2008-04-01

    Indigenous tribes and conservation biologists may have common goals and may be able to collaborate on the maintenance of biodiversity, but few researchers have evaluated the impacts and potential benefits of human subsistence activities. I studied the effects of subsistence activities (primarily wood collection) of nomadic pastoralists in 3 Afromontane forests of northern Kenya. In surveys of 404, 25-m-radius plots, I recorded vegetation structure and composition of the forest bird community. Plots with higher levels of human activity had significantly different vegetation structure, with more-open canopies, more grass, and fewer tree stems. Nectarivores (abundance +231%) and aerial insectivores (+66%) were more abundant in plots with more-intense wood collecting than in plots with less human activity, whereas abundance of forest specialists (-28%) decreased in plots with more-intense human activity. Abundance of 58% of the bird species either increased or decreased significantly in plots with more-intense human activity. Generally, the number of individuals of forest specialists decreased (6 of 7 species showed significant responses) and the number of individuals of edge and nonforest species increased with increasing human activity. Canonical correspondence analysis showed that an intensification of human activities would favor nectarivores, aerial insectivores, granivores, and omnivores and would negatively affect large-sized, ground-foraging species and arboreal frugivores. Subsistence human activities favored the invasion of forest by edge species at the expense of forest specialists; thus, further intensification of forest exploitation by local peoples is not recommended. At the same time, however, subsistence activities in northern Kenya forests appeared to increase the structural diversity of the vegetation and provided suitable habitat for part (but not all) of the forest avifauna, which suggests that subsistence human activities may have a role in the

  13. What do we know about winter active ground beetles (Coleoptera, Carabidae) in Central and Northern Europe?

    PubMed Central

    Jaskuła, Radomir; Soszyńska-Maj, Agnieszka

    2011-01-01

    Abstract This paper summarizes the current knowledge on winter active Carabidae in Central and Northern Europe. In total 73 winter active species are listed, based on literature and own observations. Ground beetles are among the three most numerous Coleoptera families active during the autumn to spring period. The winter community of Carabidae is composed both of larvae (mainly autumn breeding species) and adults, as well as of epigeic species and those inhabiting tree trunks. Supranivean fauna is characterized by lower species diversity than the subnivean fauna. The activity of ground beetles decreases in late autumn, is lowest during mid-winter and increases in early spring. Carabidae are noted as an important food source in the diet of insectivorous mammals. They are also predators, hunting small winter active invertebrates. PMID:21738431

  14. 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.

  15. Impact of variations in solar activity on hydrological decadal patterns in northern Italy

    NASA Astrophysics Data System (ADS)

    Zanchettin, D.; Rubino, A.; Traverso, P.; Tomasino, M.

    2008-06-01

    Using spectral and statistical analyses of discharges and basin average precipitation rates acquired over the Po River since the early 1800s, we investigate the impact of variations in solar activity on hydrological decadal patterns over northern Italy. Wet and dry periods appear to alternate in accordance with polarized sunspot cycles. Intriguingly, a solar signature on Po River discharges is detected to be highly significant since the late 1800s, before the onset of sunspots hyperactivity established by the middle 1900s. In particular, observed hydrological patterns over northern Italy are significantly correlated, under periods of quiet sunspot activity, with parameters characterizing the Sun's orbital motion, specifically with the time derivative of the solar angular momentum (τ) which is thought to modulate the strength of the solar wind and sunspot dynamics under weak sunspot activity. The North Atlantic Oscillation (NAO) is detected as potential link between the Sun and Po River discharges, since it is significantly correlated with both solar activity and the decadal variability in the north Italian climate. In particular, positive (negative) NAO anomalies, which are associated with comparatively lower (higher) Po River discharges, are assessed to alternatively correlate at decadal timescales either with τ or with the Earth's geomagnetic activity (GA), which closely follows sunspot activity. This changing correlation seems to be regulated by the strength of sunspot activity: under periods of quiet sunspot activity, a weakening of the GA-NAO connection and a reinforcement of the τ-NAO connection is observed. In this sense, the strength of solar activity apparently modulates the connection between the NAO and Po River discharges.

  16. Active deformation of the northern front of the Eastern Great Caucasus

    NASA Astrophysics Data System (ADS)

    Niviere, Bertrand; Gagala, Lukasz; Callot, Jean-Paul; Regard, Vincent; Ringenbach, Jean-Claude

    2016-04-01

    The Arabia-Eurasia collision involved a mosaic of island arcs and microcontinents. Their accretion to the complex paleogeographic margin of Neotethys was marked by numerous collisional events. The Greater Caucasus constitute the northernmost tectonic element of this tectonic collage, developed as a back arc extensional zone now inverted, which relationships to the onset of Arabia-Eurasia continental collision and/or to the reorganization of the Arabia-Eurasia plate boundary at ˜5 Ma remain controversial. Structurally, the Greater Caucasus are a former continental back arc rift, now the locus of ongoing continental shortening. Modern geodetic observations suggest that in the west, the strain north of the Armenian Plateau is accommodated almost exclusively along the margins of the Greater Caucasus. This differs from regions further east where strain accommodation is distributed across both the Lesser and Greater Caucasus, and within the Greater Caucasus range, with a unique southward vergence. We question here the amount and mechanisms by which the Eastern Greater Caucasus accommodate part of the Arabia-Eurasia convergence. Morphostructural analysis of the folded late Pleistocene marine terrace along the northern slope of the Eastern Greater Caucasus evidences an on going tectonic activity in the area where GPS measurements record no motion. Most of the recent foreland deformation is accommodated by south-vergent folds and thrust, i. e. opposite to the vergence of the Caucasus frontal northern thrust. A progressive unconformity in the folded beds shows that it was already active during the late Pliocene. Cosmogenic dating of the terrace and kinematic restoration of the remnant terrace, linked to the subsurface geology allows for the estimation of a shortening rate ranging from a few mm/yr to 1 cm/yr over the last 5 Myr along the greater Caucasus northern front. Thus more than one third of the shortening between the Kura block / Lesser Caucasus domain and the Stable

  17. Climate modulates internal wave activity in the Northern South China Sea

    NASA Astrophysics Data System (ADS)

    DeCarlo, Thomas M.; Karnauskas, Kristopher B.; Davis, Kristen A.; Wong, George T. F.

    2015-02-01

    Internal waves (IWs) generated in the Luzon Strait propagate into the Northern South China Sea (NSCS), enhancing biological productivity and affecting coral reefs by modulating nutrient concentrations and temperature. Here we use a state-of-the-art ocean data assimilation system to reconstruct water column stratification in the Luzon Strait as a proxy for IW activity in the NSCS and diagnose mechanisms for its variability. Interannual variability of stratification is driven by intrusions of the Kuroshio Current into the Luzon Strait and freshwater fluxes associated with the El Niño-Southern Oscillation. Warming in the upper 100 m of the ocean caused a trend of increasing IW activity since 1900, consistent with global climate model experiments that show stratification in the Luzon Strait increases in response to radiative forcing. IW activity is expected to increase in the NSCS through the 21st century, with implications for mitigating climate change impacts on coastal ecosystems.

  18. Numerical simulation of coastal flooding after potential reactivation of an active normal fault in northern Taiwan

    NASA Astrophysics Data System (ADS)

    Chan, Yu-Chang; Kuo, Chih-Yu; Chang, Kuo-Jen; Chen, Rou-Fei; Hsieh, Yu-Chung

    2016-04-01

    Rapid coastal flooding from seawards may be resulted from storm surge, tsunamis, and sudden land subsidence due to fault activities. Many observations and numerical modeling of flooding have been made for cases resulted from storm surge and tsunami events; however, coastal flooding caused by a potential normal faulting event nearby coastal areas is rarely reported. In addition to the earthquake hazards from fault rupturing and ground shaking, the accompanied hazards of earthquake-induced flooding is also important to be investigated. The Jinshan area in northern Taiwan was reported to have been flooded by a tsunami event in the year of 1867 possibly resulted from the reactivation of the Shanchiao normal fault offshore. Historical records have shown that the Shanchiao Fault that extends from Shulin along the western edge of the Taipei Basin to the town of Jinshan may have also ruptured in the year of 1694. The rupturing event has created a depression on the western side of the Taipei Basin that was later filled by sea water called the Taipei Lake. The geological conditions in northern Taiwan provide an opportunity for numerically simulating the dynamic processes of sea water flooding nearby the coastal area immediately after an earthquake-induced normal faulting event. In this study, we focused on the potential active normal faulting that may occur and result in an expected catastrophic flooding in lowland area of Jinshan in northern Taiwan. We applied the continuum shallow water equation to evaluate the unknown inundation processes including location, extent, velocity and water depths after the flooding initiated and the final state of the flooding event. The modeling results were well compared with borehole observations of the extent of previous flooding events possibly due to tsunami events. In addition, the modeling results may provide a future basis for safety evaluation of the two nuclear power plants nearby the region.

  19. 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.

  20. Monitoring and descriptive analysis of radon in relation to seismic activity of Northern Pakistan.

    PubMed

    Jilani, Zeeshan; Mehmood, Tahir; Alam, Aftab; Awais, Muhammad; Iqbal, Talat

    2017-03-16

    Earthquakes are one of the major causes of natural disasters and its forecasting is challenging task. Some precursory phenomenon exists in theory in relation to earthquakes occurrence. The emission of radioactive gas named 'radon' before the earthquakes is a potential earthquake precursory candidate. The study aims to monitor and to analyze the radon in relation to seismic activity in Northern Pakistan. For this purpose RTM-2200 has been used to monitor the changes in radon concentration from August 01, 2014 to January 31, 2015 in Northern Pakistan. Significant temporal variations has been observed in radon concentration. The bivariate analysis of radon with other variables manifests its positive relationship with air pressure and relative humidity and negative relationship with temperature. 2σ upper control limit on monthly basis are computed for detection of anomalous trends in the data. Overall increasing trend is detected in radon concentration. Five earthquakes from August 01, 2014 to January 31, 2015 have been selected from earthquake catalogue, depending upon their magnitude and distance from monitoring station and out of which radon concentration can be associated with only two earthquakes correlated with tectonic effect of radon concentration. Both of events have same magnitude 5.5 and occurred on September 13 and October 14, 2014 respectively. Very large variations have been observed in radon for the last two months of the study period, which may be occurred due to some other geological and environmental changes, but are not related to the earthquake activity.

  1. 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.

  2. Aquatic plant nutrients, moss phosphatase activities and tissue composition in four upland streams in northern England

    NASA Astrophysics Data System (ADS)

    Ellwood, N. T. W.; Haile, S. M.; Whitton, B. A.

    2008-02-01

    SummaryA study was made of the water chemistry, tissue nutrients and surface phosphatase activities of the 2-cm apices of three mosses in four upland streams in northern England, UK. This was part of a project to optimize methods for assessing nutrient fractions in environments with highly variable water chemistry. Aqueous N and P fractions showed the greatest variability followed by moss phosphatase activities, with nutrient composition of the shoot apices the least variable. There was no consistent pattern as to which aqueous N or P fraction was the most variable. The ratio between total inorganic N and total filtrable P ranged over three orders of magnitude in some streams. The interrelations between tissue N and P concentrations, tissue N:P ratio, phosphatase activities and aqueous variables showed: Significant +ve relationship between tissue N and aqueous NO 3-N in some populations, but not between tissue P and aqueous P concentration; Significant +ve relationships between phosphatase activities and aqueous organic N, but none with aqueous organic P; Significant +ve relationships between phosphodiesterase:phosphomonoesterase activities and aqueous organic N; Significant -ve relationships between phosphatase activities and tissue P concentration; Significant +ve relationships between phosphatase activities and tissue N:P. Both types of biological measurement are valuable for monitoring ambient nutrients in upland streams. Neither is clearly better than the other, so both should be included in surveys.

  3. 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.

  4. Winter climate change affects growing-season soil microbial biomass and activity in northern hardwood forests.

    PubMed

    Durán, Jorge; Morse, Jennifer L; Groffman, Peter M; Campbell, John L; Christenson, Lynn M; Driscoll, Charles T; Fahey, Timothy J; Fisk, Melany C; Mitchell, Myron J; Templer, Pamela H

    2014-11-01

    Understanding the responses of terrestrial ecosystems to global change remains a major challenge of ecological research. We exploited a natural elevation gradient in a northern hardwood forest to determine how reductions in snow accumulation, expected with climate change, directly affect dynamics of soil winter frost, and indirectly soil microbial biomass and activity during the growing season. Soils from lower elevation plots, which accumulated less snow and experienced more soil temperature variability during the winter (and likely more freeze/thaw events), had less extractable inorganic nitrogen (N), lower rates of microbial N production via potential net N mineralization and nitrification, and higher potential microbial respiration during the growing season. Potential nitrate production rates during the growing season were particularly sensitive to changes in winter snow pack accumulation and winter soil temperature variability, especially in spring. Effects of elevation and winter conditions on N transformation rates differed from those on potential microbial respiration, suggesting that N-related processes might respond differently to winter climate change in northern hardwood forests than C-related processes.

  5. A Newly Revised Master's Degree in Physical Education and Physical Activity Leadership at the University of Northern Colorado

    ERIC Educational Resources Information Center

    Dauenhauer, Brian; Krause, Jennifer; Douglas, Scott; Smith, Mark; Stellino, Megan Babkes

    2017-01-01

    In response to calls on physical education teacher education (PETE) programs to better prepare teachers for a broadened role in school-based physical activity promotion, the University of Northern Colorado has recently established a Master of Arts in teaching physical education and physical activity leadership (MAT PE-PAL) degree. The…

  6. Seasonal divergence in the interannual responses of Northern Hemisphere vegetation activity to variations in diurnal climate.

    PubMed

    Wu, Xiuchen; Liu, Hongyan; Li, Xiaoyan; Liang, Eryuan; Beck, Pieter S A; Huang, Yongmei

    2016-01-11

    Seasonal asymmetry in the interannual variations in the daytime and nighttime climate in the Northern Hemisphere (NH) is well documented, but its consequences for vegetation activity remain poorly understood. Here, we investigate the interannual responses of vegetation activity to variations of seasonal mean daytime and nighttime climate in NH (>30 °N) during the past decades using remote sensing retrievals, FLUXNET and tree ring data. Despite a generally significant and positive response of vegetation activity to seasonal mean maximum temperature (Tmax) in ~22-25% of the boreal (>50 °N) NH between spring and autumn, spring-summer progressive water limitations appear to decouple vegetation activity from the mean summer Tmax, particularly in climate zones with dry summers. Drought alleviation during autumn results in vegetation recovery from the marked warming-induced drought limitations observed in spring and summer across 24-26% of the temperate NH. Vegetation activity exhibits a pervasively negative correlation with the autumn mean minimum temperature, which is in contrast to the ambiguous patterns observed in spring and summer. Our findings provide new insights into how seasonal asymmetry in the interannual variations in the mean daytime and nighttime climate interacts with water limitations to produce spatiotemporally variable responses of vegetation growth.

  7. Screening for antibacterial and antifungal activities in marine benthic invertebrates from northern Norway.

    PubMed

    Tadesse, Margey; Gulliksen, Bjørn; Strøm, Morten B; Styrvold, Olaf B; Haug, Tor

    2008-11-01

    Benthic marine invertebrates collected from sub-Arctic regions of northern Norway, were found to be a promising source of novel bioactive compounds against human and fish pathogenic bacteria and fungi. Lyophilized material from seven species of ascidians, six sponges and one soft alcyonid coral were extracted with 60% acidified acetonitrile (ACN). After separation into an ACN-rich phase (ACN-extract) and an aqueous phase, and subsequent solid-phase extraction of the aqueous phase, fractions differing in polarity were obtained and screened for antibacterial and antifungal activities, along with the more lipophilic ACN-extracts. Antimicrobial activity was determined against two gram-negative, two gram-positive bacteria, and two strains of fungi. Notably, all the invertebrate species in the study showed activity against all four strains of bacteria and the two strains of fungi. In general, the aqueous fractions displayed highest antimicrobial activity, and the most potent extracts were obtained from the colonial ascidian Synoicum pulmonaria which displayed activity against bacteria and fungi at a concentration of 0.02 mg/ml; the lowest concentration tested.

  8. Seasonal divergence in the interannual responses of Northern Hemisphere vegetation activity to variations in diurnal climate

    PubMed Central

    Wu, Xiuchen; Liu, Hongyan; Li, Xiaoyan; Liang, Eryuan; Beck, Pieter S. A.; Huang, Yongmei

    2016-01-01

    Seasonal asymmetry in the interannual variations in the daytime and nighttime climate in the Northern Hemisphere (NH) is well documented, but its consequences for vegetation activity remain poorly understood. Here, we investigate the interannual responses of vegetation activity to variations of seasonal mean daytime and nighttime climate in NH (>30 °N) during the past decades using remote sensing retrievals, FLUXNET and tree ring data. Despite a generally significant and positive response of vegetation activity to seasonal mean maximum temperature () in ~22–25% of the boreal (>50 °N) NH between spring and autumn, spring-summer progressive water limitations appear to decouple vegetation activity from the mean summer , particularly in climate zones with dry summers. Drought alleviation during autumn results in vegetation recovery from the marked warming-induced drought limitations observed in spring and summer across 24–26% of the temperate NH. Vegetation activity exhibits a pervasively negative correlation with the autumn mean minimum temperature, which is in contrast to the ambiguous patterns observed in spring and summer. Our findings provide new insights into how seasonal asymmetry in the interannual variations in the mean daytime and nighttime climate interacts with water limitations to produce spatiotemporally variable responses of vegetation growth. PMID:26751166

  9. Western Blot of Stained Proteins from Dried Polyacrylamide Gels

    NASA Technical Reports Server (NTRS)

    Gruber, Claudia; Stan-Lotter, Helga

    1996-01-01

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

  10. Hydrochemical evaluation of the influences of mining activities on river water chemistry in central northern Mongolia.

    PubMed

    Batsaikhan, Bayartungalag; Kwon, Jang-Soon; Kim, Kyoung-Ho; Lee, Young-Joon; Lee, Jeong-Ho; Badarch, Mendbayar; Yun, Seong-Taek

    2017-01-01

    Although metallic mineral resources are most important in the economy of Mongolia, mining activities with improper management may result in the pollution of stream waters, posing a threat to aquatic ecosystems and humans. In this study, aiming to evaluate potential impacts of metallic mining activities on the quality of a transboundary river (Selenge) in central northern Mongolia, we performed hydrochemical investigations of rivers (Tuul, Khangal, Orkhon, Haraa, and Selenge). Hydrochemical analysis of river waters indicates that, while major dissolved ions originate from natural weathering (especially, dissolution of carbonate minerals) within watersheds, they are also influenced by mining activities. The water quality problem arising from very high turbidity is one of the major environmental concerns and is caused by suspended particles (mainly, sediment and soil particles) from diverse erosion processes, including erosion of river banks along the meandering river system, erosion of soils owing to overgrazing by livestock, and erosion by human activities, such as mining and agriculture. In particular, after passing through the Zaamar gold mining area, due to the disturbance of sediments and soils by placer gold mining, the Tuul River water becomes very turbid (up to 742 Nephelometric Turbidity Unit (NTU)). The Zaamar area is also the contamination source of the Tuul and Orkhon rivers by Al, Fe, and Mn, especially during the mining season. The hydrochemistry of the Khangal River is influenced by heavy metal (especially, Mn, Al, Cd, and As)-loaded mine drainage that originates from a huge tailing dam of the Erdenet porphyry Cu-Mo mine, as evidenced by δ(34)S values of dissolved sulfate (0.2 to 3.8 ‰). These two contaminated rivers (Tuul and Khangal) merge into the Orkhon River that flows to the Selenge River near the boundary between Mongolia and Russia and then eventually flows into Lake Baikal. Because water quality problems due to mining can be critical

  11. A regional reconstruction of debris-flow activity in the Northern Calcareous Alps, Austria

    NASA Astrophysics Data System (ADS)

    Procter, Emily; Bollschweiler, Michelle; Stoffel, Markus; Neumann, Mathias

    2011-09-01

    Dendrogeomorphic dating of historical debris-flow events is a highly valuable tool for improving historical records in the field of natural hazard management. Previous dendrogeomorphic investigations generally have focused on case studies of single torrents; however, regional investigations may offer a more accurate reconstruction of regional patterns of activity and therefore may have an advantage over individual cases. The aim of the study is to provide a regional reconstruction of debris-flow events for a site in the Northern Calcareous Alps of western Austria (Gamperdonatal, Vorarlberg) and to document spatial and temporal morphological changes in individual and neighboring torrents. Analysis of 442 trees (268 Pinus mugo ssp. uncinata, 164 Picea abies, and 10 Abies alba) allowed identification of 579 growth disturbances corresponding to 63 debris-flow events since A.D. 1839. The majority of growth disturbances were in the form of growth suppression or release (76%) owing to the nature of both the deposited material and the process characteristics. Regional patterns of event frequency indicated a paucity of activity in the early to mid-twentieth century and increased activity since A.D. 1948, whereby large events were followed by subsequent years of continued activity of smaller magnitude. Patterns of frequency could be attributed primarily to spatiotemporal changes in channel morphology, but may also be reflective of changes in transport conditions within the valley. This study provides the first regional investigation in the Austrian Alps and contributes to the documentation of tree responses to geomorphic disturbances in calcareous material.

  12. Numerical modeling of an intense precipitation event and its associated lightning activity over northern Greece

    NASA Astrophysics Data System (ADS)

    Pytharoulis, I.; Kotsopoulos, S.; Tegoulias, I.; Kartsios, S.; Bampzelis, D.; Karacostas, T.

    2016-03-01

    This study investigates an intense precipitation event and its lightning activity that affected northern Greece and primarily Thessaloniki on 15 July 2014. The precipitation measurement of 98.5 mm in 15 h at the Aristotle University of Thessaloniki set a new absolute record maximum. The thermodynamic analysis indicated that the event took place in an environment that could support deep thunderstorm activity. The development of this intense event was associated with significant low-level convergence and upper-level divergence even before its triggering and a positive vertical gradient of relative vorticity advection. The high resolution (1.667 km × 1.667 km) non-hydrostatic WRF-ARW numerical weather prediction model was used to simulate this intense precipitation event, while the Lightning Potential Index was utilized to calculate the potential for lightning activity. Sensitivity experiments suggested that although the strong synoptic forcing assumed primary role in the occurrence of intense precipitation and lightning activity, their spatiotemporal variability was affected by topography. The application of the very fine resolution topography of NASA Shuttle Radar Topographic Mission improved the simulated precipitation and the calculated lightning potential.

  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. Bacterial pollution, activity and heterotrophic diversity of the northern part of the Aegean Sea, Turkey.

    PubMed

    Çiftçi Türetken, Pelin S; Altuğ, Gülşen

    2016-02-01

    Isolation and characterization studies of marine heterotrophic bacteria are important to describe and understand eco-metobolism of the marine environments. In this study, diversity and community structures of the culturable heterotrophic bacteria, metabollicaly active bacteria and bacterial pollution in the coastal and offshore areas of Gökçeada Island, in the Northern Aegean Sea, Turkey were investigated from March 2012 to November 2013. The primary hydrographic parameters were recorded in situ. The frequency of the metabolically active bacteria was determined by using a modified staining technique. The indicator bacteria were determined by using membrane filtration technique; 126 bacteria isolates, 24 of them first records for this region, were identified using an automated micro-identification system, VITEK2 Compact30. The results showed that detected bacterial community profiles were significantly different when compared with previous studies conducted in polluted marine areas of Turkey. High frequency of faecal bacteria detected at station 2 indicated that increasing human activities and terrestrial pollution sources are shaping factors for possible risks, regarding recreational uses of this region, in the summer seasons.

  15. Metal contamination of active stream sediments in upper Weardale, northern Pennine Orefield, UK.

    PubMed

    Lord, R A; Morgan, P A

    2003-03-01

    In the Upper Weardale area the headwaters of the River Wear bisect the Northern Pennine Orefield, where Pb-Zn-F-Ba vein-type mineralisation has been exploited since the Roman Conquest. The area contains evidence of open pit, underground and hydraulic mining of base metal ores, associated mineral processing and smelting, exploitation of ironstones during the industrial revolution, recent extraction of fluorite and active quarrying. The aim of this study was to determine the extent of modern sediment contamination arising from these past activities. Samples of active stream sediments were collected from all major drainage channels at 1 km intervals. The sediments were analysed for Pb, Zn, Ba, Mn, Fe, Co, Ni, Cu, Cr, As, Sb, Ag and compared to data from earlier regional geochemical surveys of low order drainage samples using ArcView software. The significance of contamination levels was assessed using the Ontario aquatic sediment quality guidelines. Our results indicate widespread contamination of some major drainages by Pb, Mn, Zn and As at concentration levels anticipated to significantly affect use of the sediments by benthic organisms. Furthermore, Pb contamination shows persistence in stream sediments downstream towards agricultural areas of the floodplain and drinking water abstraction points, above which interaction with colliery mine water discharges may occur.

  16. Diet, Physical Activity, and Obesity in School-Aged Indigenous Youths in Northern Australia

    PubMed Central

    Valery, Patricia C.; Ibiebele, Torukiri; Harris, Mark; Green, Adèle C.; Cotterill, Andrew; Moloney, Aletia; Sinha, Ashim K.; Garvey, Gail

    2012-01-01

    Purpose. To examine the relationship between diet, physical activity, and obesity in Indigenous youths from northern Australia. Methods. In a cross-sectional study, physical activity and dietary intake (“short nutrition questionnaire”) were assessed among all youths during a face-to-face interview. For 92 high school youths, additional dietary information was assessed using a food-frequency questionnaire. Height and weight were measured and BMI was calculated. Multiple logistic regression was used to assess associations. Results. Of the 277 youths included, 52% had ≤2 servings of fruit and 84% had <4 servings of vegetables per day; 65% ate fish and 27%, take-away food (“fast food”) at least twice a week. One in four ate local traditional sea food including turtle and dugong (a local sea mammal) at least twice a week. Overweight/obese youths engaged in fewer days of physical activity in the previous week than normal weight youths (OR = 2.52, 95% CI 1.43–4.40), though patterns of physical activity differed by sex and age (P < 0.001). Overweight/obese youths were 1.89 times (95% CI 1.07–3.35) more likely to eat dugong regularly than nonobese youths. Analysis of food-frequency data showed no difference by weight assessment among high-school students. Conclusions. Low fruit and vegetable intake were identified in these Indigenous youths. Regular consumption of fried dugong and low frequency of physical activity were associated with overweight/obesity reinforcing the need to devise culturally appropriate health promotion strategies and interventions for Indigenous youths aimed at improving their diet and increasing their physical activity. PMID:22720140

  17. Diet, physical activity, and obesity in school-aged indigenous youths in northern australia.

    PubMed

    Valery, Patricia C; Ibiebele, Torukiri; Harris, Mark; Green, Adèle C; Cotterill, Andrew; Moloney, Aletia; Sinha, Ashim K; Garvey, Gail

    2012-01-01

    Purpose. To examine the relationship between diet, physical activity, and obesity in Indigenous youths from northern Australia. Methods. In a cross-sectional study, physical activity and dietary intake ("short nutrition questionnaire") were assessed among all youths during a face-to-face interview. For 92 high school youths, additional dietary information was assessed using a food-frequency questionnaire. Height and weight were measured and BMI was calculated. Multiple logistic regression was used to assess associations. Results. Of the 277 youths included, 52% had ≤2 servings of fruit and 84% had <4 servings of vegetables per day; 65% ate fish and 27%, take-away food ("fast food") at least twice a week. One in four ate local traditional sea food including turtle and dugong (a local sea mammal) at least twice a week. Overweight/obese youths engaged in fewer days of physical activity in the previous week than normal weight youths (OR = 2.52, 95% CI 1.43-4.40), though patterns of physical activity differed by sex and age (P < 0.001). Overweight/obese youths were 1.89 times (95% CI 1.07-3.35) more likely to eat dugong regularly than nonobese youths. Analysis of food-frequency data showed no difference by weight assessment among high-school students. Conclusions. Low fruit and vegetable intake were identified in these Indigenous youths. Regular consumption of fried dugong and low frequency of physical activity were associated with overweight/obesity reinforcing the need to devise culturally appropriate health promotion strategies and interventions for Indigenous youths aimed at improving their diet and increasing their physical activity.

  18. Increasing late winter-early spring fire activity in Northern Spain: climate change or human footprint?

    NASA Astrophysics Data System (ADS)

    Carracedo Martín, Virginia; García Codron, Juan Carlos; Rasilla Álvarez, Domingo

    2016-04-01

    Most of the fire activity across Spain concentrates during the summer months, but a secondary peak appears also during late winter and early spring (February and March). This peak represents a tiny fraction of the burned surface but in northern Spain becomes the main fire season, representing up to 60 % of the total burned surface. Moreover, the impact of this "unseasonal" fire regime is becoming more relevant; an analysis of the temporal evolution of the burned surface since 2005 shows that the suppression efforts of summer forest fires have apparently succeeded, while the opposite has occurred with late winter-early spring forest fires. For example, during March 2012 more than 22,000 ha were burned in the Spanish provinces of Asturias and Cantabria, while about 14,000 suffers the effects of fires in Northern Portugal. Anthropogenic factor (mostly linked to an extensive cattle farming in the mountains) are the main cause of such fire activity, but atmospheric factors also play a relevant role in the spread of this fires. Consequently, the main aim of this poster is to explore if the recent evolution of forest fires in the study area are consequence of an aggravation of the atmospheric conditions driving to more fire risk conditions, or other factor could also explain the increase in fire activity. Burned surface data obtained from official statistics since 1971 were compared with atmospheric data at two temporal scales: daily fire risk values calculated from synoptic records and long term drought indices (SPI and SPEI). The results show a long term increase in both daily fire risk and drought conditions, but this trend can be related to the background warming of the area, rather to an increase in the frequency and magnitude of the extreme fire weather events. Thus, we consider that the regional atmospheric evolution cannot explain by itself the recent increase in late winter-early spring fire activity. Additional anthropogenic factors, such as recent changes in

  19. No covariation between the geomagnetic activity and the incidence of acute myocardial infarction in the polar area of northern Sweden.

    PubMed

    Messner, T; Häggström, I; Sandahl, I; Lundberg, V

    2002-05-01

    This study was undertaken to investigate whether there was any relation between the aurora borealis (measured as the geomagnetic activity) and the number of acute myocardial infarctions (AMI) in the northern, partly polar, area of Sweden. The AMI cases were collected from The Northern Sweden MONICA (multinational MONItoring of trends and determinants of CArdiovascular disease) AMI registry between 1985 and 1998, inclusive, and the information on the geomagnetic activity from continuous measurements at the Swedish Institute of Space Physics, Kiruna. In the analyses, both the relation between the individual AMI case and ambient geomagnetic activity, and the relation between the mean daily K index and the daily number of AMI cases were tested. We found no statistically significant relation between the number of fatal or non-fatal AMI cases, the number of sudden deaths or the number of patients with chest pain without myocardial damage, and geomagnetic activity. Our data do not support a relation between the geomagnetic activity and AMI.

  20. No covariation between the geomagnetic activity and the incidence of acute myocardial infarction in the polar area of northern Sweden

    NASA Astrophysics Data System (ADS)

    Messner, T.; Häggström, I.; Sandahl, I.; Lundberg, V.

    2002-05-01

    This study was undertaken to investigate whether there was any relation between the aurora borealis (measured as the geomagnetic activity) and the number of acute myocardial infarctions (AMI) in the northern, partly polar, area of Sweden. The AMI cases were collected from The Northern Sweden MONICA (multinational MONItoring of trends and determinants of CArdiovascular disease) AMI registry between 1985 and 1998, inclusive, and the information on the geomagnetic activity from continuous measurements at the Swedish Institute of Space Physics, Kiruna. In the analyses, both the relation between the individual AMI case and ambient geomagnetic activity, and the relation between the mean daily K index and the daily number of AMI cases were tested. We found no statistically significant relation between the number of fatal or non-fatal AMI cases, the number of sudden deaths or the number of patients with chest pain without myocardial damage, and geomagnetic activity. Our data do not support a relation between the geomagnetic activity and AMI.

  1. Assessing Health-Related Quality of Life in Northern Plains American Indians: Prominence of Physical Activity as a Health Behavior

    ERIC Educational Resources Information Center

    Poltavski, Dmitri; Holm, Jeffrey; Vogeltanz-Holm, Nancy; McDonald, Leander

    2010-01-01

    Associations of behavioral health risks and healthy behaviors with self-reported health-related quality of life measures were investigated in a Northern Plains American Indian sample. Participants were surveyed in person using the Behavioral Risk Factor Surveillance Survey. The results showed that regular physical activity was significantly…

  2. The Impact of the School-Based Psychosocial Structured Activities (PSSA) Program on Conflict-Affected Children in Northern Uganda

    ERIC Educational Resources Information Center

    Ager, Alastair; Akesson, Bree; Stark, Lindsay; Flouri, Eirini; Okot, Braxton; McCollister, Faith; Boothby, Neil

    2011-01-01

    Background: Children in northern Uganda have undergone significant psychosocial stress during the region's lengthy conflict. A Psychosocial Structured Activities (PSSA) program was implemented in 21 schools identified as amongst those most severely affected by conflict-induced displacement across Gulu and Amuru Districts. The PSSA intervention…

  3. High-resolution imagery of active faulting offshore Al Hoceima, Northern Morocco

    NASA Astrophysics Data System (ADS)

    d'Acremont, E.; Gutscher, M.-A.; Rabaute, A.; Mercier de Lépinay, B.; Lafosse, M.; Poort, J.; Ammar, A.; Tahayt, A.; Le Roy, P.; Smit, J.; Do Couto, D.; Cancouët, R.; Prunier, C.; Ercilla, G.; Gorini, C.

    2014-09-01

    Two recent destructive earthquakes in 1994 and 2004 near Al Hoceima highlight that the northern Moroccan margin is one of the most seismically active regions of the Western Mediterranean area. Despite onshore geodetic, seismological and tectonic field studies, the onshore-offshore location and extent of the main active faults remain poorly constrained. Offshore Al Hoceima, high-resolution seismic reflection and swath-bathymetry have been recently acquired during the Marlboro-2 cruise. These data at shallow water depth, close to the coast, allow us to describe the location, continuity and geometry of three active faults bounding the offshore Nekor basin. The well-expressed normal-left-lateral onshore Trougout fault can be followed offshore during several kilometers with a N171°E ± 3° trend. Westward, the Bousekkour-Aghbal normal-left-lateral onshore fault is expressed offshore with a N020°E ± 4° trending fault. The N030°E ± 2° Bokkoya fault corresponds to the western boundary of the Plio-Quaternary offshore Nekor basin in the Al Hoceima bay and seems to define an en échelon tectonic pattern with the Bousekkour-Aghbal fault. We propose that these three faults are part of the complex transtensional system between the Nekor fault and the Al-Idrissi fault zone. Our characterization of the offshore expression of active faulting in the Al Hoceima region is consistent with the geometry and nature of the active fault planes deduced from onshore geomorphological and morphotectonic analyses, as well as seismological, geodetic and geodynamic data.

  4. Measurements of 222Rn activity concentration in domestic water sources in Penang, northern peninsular Malaysia.

    PubMed

    Muhammad, B G; Jaafar, M S; Azhar, A R; Akpa, T C

    2012-04-01

    Measurements of (222)Rn activity concentration were carried out in 39 samples collected from the domestic and drinking water sources used in the island and mainland of Penang, northern peninsular, Malaysia. The measured activity concentrations ranged from 7.49 to 26.25 Bq l(-1), 0.49 to 9.72 Bq l(-1) and 0.58 to 2.54 Bq l(-1) in the raw, treated and bottled water samples collected, respectively. This indicated relatively high radon concentrations compared with that from other parts of the world, which still falls below the WHO recommended treatment level of 100 Bq l(-1). From this data, the age-dependent associated committed effective doses due to the ingestion of (222)Rn as a consequence of direct consumption of drinking water were calculated. The committed effective doses from (222)Rn resulting from 1 y's consumption of these water were estimated to range from 0.003 to 0.048, 0.001 to 0.018 and 0.002 to 0.023 mSv y(-1), for age groups 0-1, 2-16 and >16 y, respectively.

  5. Major off-axis hydrothermal activity on the northern Gorda Ridge

    USGS Publications Warehouse

    Rona, Peter A.; Denlinger, Roger P.; Fisk, M. R.; Howard, K. J.; Taghon, G. L.; Klitgord, Kim D.; McClain, James S.; McMurray, G. R.; Wiltshire, J. C.

    1990-01-01

    The first hydrothermal field on the northern Gorda Ridge, the Sea Cliff hydrothermal field, was discovered and geologic controls of hydrothermal activity in the rift valley were investigated on a dive series using the DSV Sea Cliff. The Sea Cliff hydrothermal field was discovered where predicted at the intersection of axis-oblique and axis-parallel faults at the south end of a linear ridge at mid-depth (2700 m) on the east wall. Preliminary mapping and smpling of the field reveal: a setting nested on nearly sediment-free fault blocks 300 m above the rift valley floor 2.6 km from the axis; a spectrum of venting types from seeps to black smokers; high conductive heat flow estimated to be equivalent to the convective flux of multiple black smokers through areas of the sea floor sealed by a caprock of elastic breccia primarily derived from basalt with siliceous cement and barite pore fillings; and a vent biota with Juan de Fuca Ridge affinites. These findings demonstrate the importance of off-axis hydrothermal activity and the role of the intersection of tectonic lineations in controlling hydrothermal sites at sea-floor spreading centers.

  6. The criminal use of improvised and re-activated firearms in Great Britain and Northern Ireland.

    PubMed

    Warlow, Thomas A

    2007-11-01

    Following the Hungerford Massacre the British Government imposed additional strict firearms legislation in 1988 that involved several classes of arms being placed into the prohibited category. By way of compensation a scheme was introduced to allow the unrestricted possession and transfer of the newly prohibited arms, other firearms and "prohibited weapons", if de-activated to a standard acceptable to the Secretary of State. Approved standards for firearm de-activation were drawn up in 1989. The inspection of the de-activated arms was devolved to the two Gun Barrel Proof Houses at London and Birmingham, as otherwise the task of inspecting the thousands of guns involved would have overloaded the firearms section at the Huntingdon Forensic Science Service Laboratory, who were already dealing with criminal firearms cases submitted by 41 of the 43 police forces throughout England and Wales, as well as providing technical assistance to the Home Office and Government Ministers. Members of the Gun Trade made representations to the Minister involved during the initial stages of setting up the official de-activation standards. This resulted in some measure of compromise in the range and nature of the de-activation requirements. Although it was clear that some individuals possessing the necessary skill and equipment might attempt to restore the odd weapon to a working condition, the scheme appeared to work reasonably well for the next few years. However, over the passage of time, criminal casework submissions to the Huntingdon Laboratory from industrial city areas along the M62 corridor of northern England revealed a steadily growing trend in the use of re-activated arms, which in a significant number of cases involved the use of fully automatic weapons. At first, the nature of the re-activation processes used to restore these arms was quite crude. However, with the passage of time a steady improvement in the machining and welding skills used by the some of the culprits

  7. Seismic surveys negatively affect humpback whale singing activity off northern Angola.

    PubMed

    Cerchio, Salvatore; Strindberg, Samantha; Collins, Tim; Bennett, Chanda; Rosenbaum, Howard

    2014-01-01

    Passive acoustic monitoring was used to document the presence of singing humpback whales off the coast of Northern Angola, and opportunistically test for the effect of seismic survey activity in the vicinity on the number of singing whales. Two Marine Autonomous Recording Units (MARUs) were deployed between March and December 2008 in the offshore environment. Song was first heard in mid June and continued through the remaining duration of the study. Seismic survey activity was heard regularly during two separate periods, consistently throughout July and intermittently in mid-October/November. Numbers of singers were counted during the first ten minutes of every hour for the period from 24 May to 1 December, and Generalized Additive Mixed Models (GAMMs) were used to assess the effect of survey day (seasonality), hour (diel variation), moon phase and received levels of seismic survey pulses (measured from a single pulse during each ten-minute sampled period) on singer number. Application of GAMMs indicated significant seasonal variation, which was the most pronounced effect when assessing the full dataset across the entire season (p<0.001); however seasonality almost entirely dropped out of top-ranked models when applied to a reduced dataset during the July period of seismic survey activity. Diel variation was significant in both the full and reduced datasets (from p<0.01 to p<0.05) and often included in the top-ranked models. The number of singers significantly decreased with increasing received level of seismic survey pulses (from p<0.01 to p<0.05); this explanatory variable was included among the top ranked models for one MARU in the full dataset and both MARUs in the reduced dataset. This suggests that the breeding display of humpback whales is disrupted by seismic survey activity, and thus merits further attention and study, and potentially conservation action in the case of sensitive breeding populations.

  8. Natural oil slicks fuel surface water microbial activities in the northern Gulf of Mexico.

    PubMed

    Ziervogel, Kai; D'Souza, Nigel; Sweet, Julia; Yan, Beizhan; Passow, Uta

    2014-01-01

    We conducted a series of roller tank incubations with surface seawater from the Green Canyon oil reservoir, northern Gulf of Mexico, amended with either a natural oil slick (GCS-oil) or pristine oil. The goal was to test whether bacterial activities of natural surface water communities facilitate the formation of oil-rich marine snow (oil snow). Although oil snow did not form during any of our experiments, we found specific bacterial metabolic responses to the addition of GCS-oil that profoundly affected carbon cycling within our 4-days incubations. Peptidase and β-glucosidase activities indicative of bacterial enzymatic hydrolysis of peptides and carbohydrates, respectively, were suppressed upon the addition of GCS-oil relative to the non-oil treatment, suggesting that ascending oil and gas initially inhibits bacterial metabolism in surface water. Biodegradation of physically dispersed GCS-oil components, indicated by the degradation of lower molecular weight n-alkanes as well as the rapid transformation of particulate oil-carbon (C: N >40) into the DOC pool, led to the production of carbohydrate- and peptide-rich degradation byproducts and bacterial metabolites such as transparent exopolymer particles (TEP). TEP formation was highest at day 4 in the presence of GCS-oil; in contrast, TEP levels in the non-oil treatment already peaked at day 2. Cell-specific enzymatic activities closely followed TEP concentrations in the presence and absence of GCS-oil. These results demonstrate that the formation of oil slicks and activities of oil-degrading bacteria result in a temporal offset of microbial cycling of organic matter, affecting food web interactions and carbon cycling in surface waters over cold seeps.

  9. Natural oil slicks fuel surface water microbial activities in the northern Gulf of Mexico

    PubMed Central

    Ziervogel, Kai; D'souza, Nigel; Sweet, Julia; Yan, Beizhan; Passow, Uta

    2014-01-01

    We conducted a series of roller tank incubations with surface seawater from the Green Canyon oil reservoir, northern Gulf of Mexico, amended with either a natural oil slick (GCS-oil) or pristine oil. The goal was to test whether bacterial activities of natural surface water communities facilitate the formation of oil-rich marine snow (oil snow). Although oil snow did not form during any of our experiments, we found specific bacterial metabolic responses to the addition of GCS-oil that profoundly affected carbon cycling within our 4-days incubations. Peptidase and β-glucosidase activities indicative of bacterial enzymatic hydrolysis of peptides and carbohydrates, respectively, were suppressed upon the addition of GCS-oil relative to the non-oil treatment, suggesting that ascending oil and gas initially inhibits bacterial metabolism in surface water. Biodegradation of physically dispersed GCS-oil components, indicated by the degradation of lower molecular weight n-alkanes as well as the rapid transformation of particulate oil-carbon (C: N >40) into the DOC pool, led to the production of carbohydrate- and peptide-rich degradation byproducts and bacterial metabolites such as transparent exopolymer particles (TEP). TEP formation was highest at day 4 in the presence of GCS-oil; in contrast, TEP levels in the non-oil treatment already peaked at day 2. Cell-specific enzymatic activities closely followed TEP concentrations in the presence and absence of GCS-oil. These results demonstrate that the formation of oil slicks and activities of oil-degrading bacteria result in a temporal offset of microbial cycling of organic matter, affecting food web interactions and carbon cycling in surface waters over cold seeps. PMID:24847314

  10. Seismicity characteristics of a potentially active Quaternary volcano: The Tatun Volcano Group, northern Taiwan

    NASA Astrophysics Data System (ADS)

    Konstantinou, Konstantinos I.; Lin, Cheng-Horng; Liang, Wen-Tzong

    2007-02-01

    The Tatun Volcano Group (TVG) is located at the northern tip of Taiwan, near the capital Taipei and close to two nuclear power plants. Because of lack of any activity in historical times it has been classified as an extinct volcano, even though more recent studies suggest that TVG might have been active during the last 20 ka. In May 2003 a seismic monitoring project at the TVG area was initiated by deploying eight three-component seismic stations some of them equipped with both short-period and broadband sensors. During the 18 months observation period local seismicity mainly consisted of high frequency earthquakes either occurring as isolated events, or as a continuous sequence in the form of spasmodic bursts. Mixed and low frequency events were also present during the same period, even though they occurred only rarely. Arrival times from events with clear P-/S-wave phases were inverted in order to obtain a minimum 1D velocity model with station corrections. Probabilistic nonlinear earthquake locations were calculated for all these events using the newly derived velocity model. Most high frequency seismicity appeared to be concentrated near the areas of hydrothermal activity, forming tight clusters at depths shallower than 4 km. Relative locations, calculated using the double-difference method and utilising catalogue and cross-correlation differential traveltimes, showed insignificant differences when compared to the nonlinear probabilistic locations. In general, seismicity in the TVG area seems to be primarily driven by circulation of hydrothermal fluids as indicated by the occurrence of spasmodic bursts, mixed/low frequency events and a b-value (1.17 ± 0.1) higher than in any other part of Taiwan. These observations, that are similar to those reported in other dormant Quaternary volcanoes, indicate that a magma chamber may still exist beneath TVG and that a future eruption or period of unrest should not be considered unlikely.

  11. Bacterial biomass and activity in the marginal ice zone of the northern Barents Sea

    NASA Astrophysics Data System (ADS)

    Tammert, Helen; Olli, Kalle; Sturluson, Maria; Hodal, Helene

    2008-10-01

    Bacteria in the Arctic Waters are well adapted to low temperatures and play a key role in the transformation of organic matter. However, the activity of planktonic bacteria at cellular level remains poorly understood. In this study, we use fluorescent markers (4',6'-diamidino-2-phenylindole (DAPI), 5-cyano-2,3-ditolyl tetrazolium chloride (CTC), Live/Dead BacLight viability kit) to discriminate between bacterial cells with a variety of physiological activities in the 0-200 m water column and sinking particles. During two field studies (July 2003 and 2004), we covered nine stations in the northern Barents Sea. The median bacterial abundance (DAPI staining) in the upper 50 m layer was 0.9×10 6 cells ml -1 (range 0.2-3.2×10 6 cells ml -1) in 2003 and 0.5×10 6 cells ml -1 (range 0.2-1.0×10 6 cells ml -1) in 2004. Bacteria with sufficient electron transport activity to be stained with CTC were on average 10% of the total count and ca. 20% of the total cells had intact cell membranes. In the water column, proxies of substrate availability (POC, PON, chlorophyll a, primary production) and bacterial production (thymidine and leucine uptake) correlated strongly with total bacterial count, CTC-stained cells and cells with 'leaky' membrane (stained with propidium iodine), but not with the concentration of cells with intact cell membrane. Contrary to expectations, the proportion of CTC-stained bacteria was not higher in the sinking particles (captured with sediment traps) compared to the ambient water. However, out of the bacteria with intact cell membranes, a higher proportion scored as CTC positive in the aggregates compared to the ambient water. Bacterial cells with 'leaky' cell membranes formed the largest part of total cell count in all samples, and accumulated in sites with high microbial activity (sinking aggregates, chlorophyll maxima, layers of high primary and bacterial production). We hypothesize that the source of the bacterial cells with 'leaky' cell membranes

  12. Active landsliding and landscape denudation in response to transient tectonic uplift, Northern California.

    NASA Astrophysics Data System (ADS)

    Bennett, G. L.; Roering, J. J.; Miller, S. R.; Kirby, E.; Schmidt, D. A.

    2014-12-01

    The northern Californian Coast ranges present a unique area to study landscape response to transient tectonic uplift. Studies have shown that an increase in uplift may be balanced by the rate of landsliding in settings of steady uplift. However, the landsliding response to transient tectonic uplift remains to be elucidated. The Californian Coast ranges are shaped by the northward migration of the Mendocino Triple Junction (MTJ), which geodynamic modeling suggests produces a transient double-humped uplift field. A major research question is whether we can detect a signature of this transient tectonic uplift in landslide activity and document how the channel network communicates this signal to hillslopes. Using air photos and Worldview imagery, we manually mapped more than 2000 earthflows and debris slides in the Eel and surrounding catchments that span the ~400 km-long region. The velocities of active earthflows were estimated by visually tracking features between images spanning 1993 to 2013. We mapped channel steepness from 10m NED DEMs in Topotoolbox 2 and developed a new tool to automatically define knickpoints along the channel network. Earthflows occur almost exclusively in a band of Franciscan mélange oriented along the MTJ transect whilst debris slides are more evenly distributed by lithology. Both earthflows and debris slides are clustered in the Eel catchment around the proposed uplift peaks and are largely absent outside of these zones. Within these areas of high landslide densities, we observe peaks in active earthflows adjacent to peaks in dormant earthflows to the south, suggesting that the signature of earthflow activity remains for a period of time once the uplift peak has passed. Landslide density, mean landslide area, and earthflow velocity all increase rapidly above threshold values of channel steepness and local relief. In the Eel catchment, where the zone of rapid uplift is commencing, landslides, particularly earth flows, are concentrated

  13. Extracellular enzyme activity and dynamics of bacterial community in mucilaginous aggregates of the northern Adriatic Sea.

    PubMed

    Zoppini, Annamaria; Puddu, Alberto; Fazi, Stefano; Rosati, Michela; Sist, Paola

    2005-12-15

    Bacterial degradation of mucilaginous aggregates (creamy layers, stringers and macroflocs) collected during two summer events (2001-2002) was tested. The objective was to describe the temporal trend of the bacterial activity, abundance and composition in the aggregated and dissolved organic matter under different trophic conditions. In the native aggregates proteins and organic phosphorous were actively hydrolyzed as aminopeptidase and alkaline phosphatase activities represented up to 87% and 25% of total activity, respectively; polysaccharides were less hydrolyzed and the highest activities were observed for beta-glucosidase (5% of the total). This hydrolysation pattern tends to a progressive accumulation of long persistent polysaccharides. During short term incubations nutrient addition (P, N and Glucose) differently stimulated bacterial growth in the seawater: P played the main role in stimulating bacterial production from 3 to 6 folds higher than in the control, whereas a secondary C-limitation was observed only for bacteria growing on seawater from macroflocs. This scarce dissolved organic carbon (DOC) bioavailability was confirmed by the lower DOC removal (13% macroflocs, 36% stringers). The total amount of carbon incorporated by bacteria living on aggregates was similar (0.58 mg C L(-1)) both in the control and under P enrichments showing a more balanced condition with respect to the seawater. Hence the well-known P limitation in the Northern Adriatic Sea affects only dissolved organic carbon uptake without influencing the uptake of aggregated organic matter. Organic matter limitation was observed only on stringers--total C incorporated raised to 0.96 mg C L(-1) after PNG addition. Macroflocs release of refractory compounds leads to DOC accumulation (73 microM DOC) contributing to inflate the pool of refractory DOC in the surrounding waters. Several evidences, including different monosaccharide composition of stringers and macroflocs (glucose 15% and 56% on

  14. Climate impacts on human settlement and agricultural activities in northern Norway revealed through sediment biogeochemistry.

    PubMed

    D'Anjou, Robert M; Bradley, Raymond S; Balascio, Nicholas L; Finkelstein, David B

    2012-12-11

    Disentangling the effects of climate change and anthropogenic activities on the environment is a major challenge in paleoenvironmental research. Here, we used fecal sterols and other biogeochemical compounds in lake sediments from northern Norway to identify both natural and anthropogenic signals of environmental change during the late Holocene. The area was first occupied by humans and their grazing animals at ∼2,250 ± 75 calendar years before 1950 AD (calendar years before present). The arrival of humans is indicated by an abrupt increase in coprostanol (and its epimer epicoprostanol) in the sediments and an associated increase in 5β-stigmastanol (and 5β-epistigmastanol), which resulted from human and animal feces washing into the lake. Human settlement was accompanied by an abrupt increase in landscape fires (indicated by the rise in pyrolytic polycyclic aromatic hydrocarbons) and a decline in woodland (registered by a change in n-alkane chain lengths from leaf waxes), accelerating a process that began earlier in the Holocene. Human activity and associated landscape changes in the region over the last two millennia were mainly driven by summer temperatures, as indicated by independent tree-ring reconstructions, although there were periods when socioeconomic factors played an equally important role. In this study, fecal sterols in lake sediments have been used to provide a record of human occupancy through time. This approach may be useful in many archeological studies, both to confirm the presence of humans and grazing animals, and to distinguish between anthropogenic and natural factors that have influenced the environment in the past.

  15. 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.

  16. Composition, size and cloud condensation nuclei activity of biomass burning aerosol from northern Australian savannah fires

    NASA Astrophysics Data System (ADS)

    Mallet, Marc D.; Cravigan, Luke T.; Milic, Andelija; Alroe, Joel; Ristovski, Zoran D.; Ward, Jason; Keywood, Melita; Williams, Leah R.; Selleck, Paul; Miljevic, Branka

    2017-03-01

    variations in the activation diameters. Trade winds carry the smoke generated from these fires over the Timor Sea, where aerosol-cloud interactions are likely to be sensitive to changes in CCN concentrations, perturbing cloud albedo and lifetime. Dry season fires in northern Australia are therefore potentially very important in cloud processes in this region.

  17. Geomorphic signal of active faulting at the northern edge of Lut Block: Insights on the kinematic scenario of Central Iran

    NASA Astrophysics Data System (ADS)

    Calzolari, Gabriele; Della Seta, Marta; Rossetti, Federico; Nozaem, Reza; Vignaroli, Gianluca; Cosentino, Domenico; Faccenna, Claudio

    2016-01-01

    Recent works documented Neogene to Quaternary dextral strike-slip tectonics along the Kuh-e-Sarhangi and Kuh-e-Faghan intraplate strike-slip faults at the northern edge of the Lut Block of Central Iran, previously thought to be dominated by sinistral strike-slip deformation. This work focuses on the evidence of Quaternary activity of one of these fault systems, in order to provide new spatiotemporal constraints on their role in the active regional kinematic scenario. Through geomorphological and structural investigation, integrated with optically stimulated luminescence dating of three generations of alluvial fans and fluvial terraces (at ~53, ~25, and ~6 ka), this study documents (i) the topographic inheritance of the long-term (Myr) punctuated history of fault nucleation, propagation, and exhumation along the northern edge of Lut Block; (ii) the tectonic control on drainage network evolution, pediment formation, fluvial terraces, and alluvial fan architecture; (iii) the minimum Holocene age of Quaternary dextral strike-slip faulting; and (iv) the evidence of Late Quaternary fault-related uplift localized along the different fault strands. The documented spatial and temporal constraints on the active dextral strike-slip tectonics at the northern edge of Lut Block provide new insights on the kinematic model for active faulting in Central Iran, which has been reinterpreted in an escape tectonic scenario.

  18. 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.

  19. 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

  20. Seasonally different response of photosynthetic activity to daytime and night-time warming in the Northern Hemisphere

    DOE PAGES

    Tan, Jianguang; Piao, Shilong; Chen, Anping; ...

    2014-08-27

    Over the last century the Northern Hemisphere has experienced rapid climate warming, but this warming has not been evenly distributed seasonally, as well as diurnally. The implications of such seasonal and diurnal heterogeneous warming on regional and global vegetation photosynthetic activity, however, are still poorly understood. Here, we investigated for different seasons how photosynthetic activity of vegetation correlates with changes in seasonal daytime and night-time temperature across the Northern Hemisphere (>30°N), using Normalized Difference Vegetation Index (NDVI) data from 1982 to 2011 obtained from the Advanced Very High Resolution Radiometer (AVHRR). Our analysis revealed some striking seasonal differences in themore » response of NDVI to changes in day- versus night-time temperatures. For instance, while higher daytime temperature (Tmax) is generally associated with higher NDVI values across the boreal zone, the area exhibiting a statistically significant positive correlation between Tmax and NDVI is much larger in spring (41% of area in boreal zone – total area 12.6 × 106 km2) than in summer and autumn (14% and 9%, respectively). In contrast to the predominantly positive response of boreal ecosystems to changes in Tmax, increases in Tmax tended to negatively influence vegetation growth in temperate dry regions, particularly during summer. Changes in night-time temperature (Tmin) correlated negatively with autumnal NDVI in most of the Northern Hemisphere, but had a positive effect on spring and summer NDVI in most temperate regions (e.g., Central North America and Central Asia). Such divergent covariance between the photosynthetic activity of Northern Hemispheric vegetation and day- and night-time temperature changes among different seasons and climate zones suggests a changing dominance of ecophysiological processes across time and space. Lastly, understanding the seasonally different responses of vegetation photosynthetic activity to diurnal

  1. Prokaryotic Abundance and Activity in Permafrost of the Northern Victoria Land and Upper Victoria Valley (Antarctica).

    PubMed

    La Ferla, Rosabruna; Azzaro, Maurizio; Michaud, Luigi; Caruso, Gabriella; Lo Giudice, Angelina; Paranhos, Rodolfo; Cabral, Anderson S; Conte, Antonella; Cosenza, Alessandro; Maimone, Giovanna; Papale, Maria; Rappazzo, Alessandro Ciro; Guglielmin, Mauro

    2017-03-13

    Victoria Land permafrost harbours a potentially large pool of cold-affected microorganisms whose metabolic potential still remains underestimated. Three cores (BC-1, BC-2 and BC-3) drilled at different depths in Boulder Clay (Northern Victoria Land) and one sample (DY) collected from a core in the Dry Valleys (Upper Victoria Valley) were analysed to assess the prokaryotic abundance, viability, physiological profiles and potential metabolic rates. The cores drilled at Boulder Clay were a template of different ecological conditions (different temperature regime, ice content, exchanges with atmosphere and with liquid water) in the same small basin while the Dry Valleys site was very similar to BC-2 conditions but with a complete different geological history and ground ice type. Image analysis was adopted to determine cell abundance, size and shape as well as to quantify the potential viable and respiring cells by live/dead and 5-cyano-2,3-ditolyl-tetrazolium chloride staining, respectively. Subpopulation recognition by apparent nucleic acid contents was obtained by flow cytometry. Moreover, the physiological profiles at community level by Biolog-Ecoplate™ as well as the ectoenzymatic potential rates on proteinaceous (leucine-aminopeptidase) and glucidic (ß-glucosidase) organic matter and on organic phosphates (alkaline-phosphatase) by fluorogenic substrates were tested. The adopted methodological approach gave useful information regarding viability and metabolic performances of microbial community in permafrost. The occurrence of a multifaceted prokaryotic community in the Victoria Land permafrost and a large number of potentially viable and respiring cells (in the order of 10(4)-10(5)) were recognised. Subpopulations with a different apparent DNA content within the different samples were observed. The physiological profiles stressed various potential metabolic pathways among the samples and intense utilisation rates of polymeric carbon compounds and carbohydrates

  2. Camera trap records of animal activity prior to a M=7 earthquake in Northern Peru

    NASA Astrophysics Data System (ADS)

    Grant, R.; Raulin, J.; Freund, F.

    2013-12-01

    Earthquake (EQ) preparation is associated with geophysical changes occurring over many scales. Some pre-earthquake (pre-EQ) processes affect the ionosphere, others leave their mark on biota. We report (i) on ionospheric anomalies recorded prior to the M=7 Contamana EQ [1] in North-Eastern Peru, 134 km deep, associated with the subduction of the Nazca plate underneath the Northern Andes, (ii) on changes in animal activity recorded in the Yanachaga National Park, about 320 km from the EQ epicentre, over a 30 day period leading up to the M=7 seismic event. Night-time Very Low Frequency (VLF) phase data were analyzed for the period 01 June to 31 Oct. 2011 using propagation paths passing close to the Yanachaga Park from the NAA emitter (USA) to receivers PIU in Piura and PLO in Lima (Peru). Ionospheric phase perturbations were observed starting 2 weeks before the EQ with periodicities from few tens of secs to few minutes. Animal activity data were obtained by evaluating the images of a cluster of 10 motion-triggered cameras of the Tropical Ecology Assessment and Monitoring Network www.teamnetwork.org. We analyzed 1359 photographic records for the pre-EQ period and 1491 photographic records for a control period with low seismicity. Animal activity started to noticeably decline 3 weeks before the EQ. Different animal species were found to react differently. The number of rodents declined to zero about one week before the EQ and so did the number of tapirs. Armadillos, a burrowing animal, were recorded in larger numbers. Though the armadillos were presumably also flushed out of their holes, they apparently did not hide like the rodents. We discuss the results in the context of recent advances in solid state physics, which provide plausible mechanisms for pre-EQ ionospheric anomalies and for changes in animal behavior. [1] Tavera, H. (2012), Report on the 24 Aug. 2011 M 7.0 Contamana, Peru, Intermediate Depth Earthquake Seismological Research Letters, 83, 1007-1013, doi: 10.1785/0220120005

  3. Urban Geocryology: Mapping Urban-Rural Contrasts in Active-Layer Thickness, Barrow Penninsula, Northern Alaska

    NASA Astrophysics Data System (ADS)

    Klene, A. E.; Nelson, F. E.

    2014-12-01

    As development proceeds in the high latitudes, information about interactions between urban influences and the thickness of the active layer above permafrost becomes vital, particularly given the possibility of increasing temperatures accompanying climate change. Permafrost characteristics are often mapped at small geographical scales (i.e., over large areas), at low resolution, and without extensive field validation. Although maps of active-layer thickness (ALT) have been created for areas of relatively undisturbed terrain, this has rarely been done within urbanized areas, even though ALT is a critical factor in the design of roads, buildings, pipelines, and other elements of infrastructure. The need for detailed maps of ALT is emphasized in work on potential hazards in permafrost regions associated with global warming scenarios. Northern Alaska is a region considered to be at moderate to high risk for thaw-induced damage under climatic warming. The Native Village of Barrow (71°17'44"N; 156°45' 59"W), the economic, transportation, and administrative hub of the North Slope Borough, is the northernmost community in the USA, and the largest native settlement in the circum-Arctic. A winter urban heat island in Barrow, earlier snowmelt in the village, and dust deposition downwind of gravel pads and roads are all urban effects that could increase ALT. A recent empirical study documented a 17 to 41 cm difference in ALT between locations in the village of Barrow and surrounding undeveloped tundra, even in similar land-cover classes. We mapped ALT in the Barrow Peninsula, with particular attention to contrasts between the urbanized village and relatively undisturbed tundra in the nearby Barrow Environmental Observatory. The modified Berggren solution, an advanced analytic solution to the general Stefan problem of calculating frost and thaw depth, was used in a geographic context to map ALT over the 150 km² area investigated in the Barrow Urban Heat Island Study. The

  4. Azithromycin Activities against Orientia tsutsugamushi Strains Isolated in Cases of Scrub Typhus in Northern Thailand

    PubMed Central

    Watt, George; Kantipong, Pacharee; Jongsakul, Krisada; Watcharapichat, Pochaman; Phulsuksombati, Duangporn

    1999-01-01

    Azithromycin was given to mice and humans infected with strains of Orientia tsutsugamushi from northern Thailand, where drug-resistant scrub typhus occurs. Azithromycin and doxycycline yielded comparable mouse survival rates (73 and 79%, respectively; P > 0.5). Symptoms, signs, and fever in two pregnant women abated rapidly with azithromycin. Prospective human trials are needed. PMID:10543774

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

    PubMed Central

    2015-01-01

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

  6. Historical record of nuclear activities from (129)I in corals from the northern hemisphere (Philippines).

    PubMed

    Bautista, Angel T; Matsuzaki, Hiroyuki; Siringan, Fernando P

    2016-11-01

    Iodine-129 is a long-lived fission product that is majorly released in human nuclear activities (HNA) such as nuclear bomb testing, nuclear fuel reprocessing, and nuclear accidents. It is a good environmental tracer and former measurements of (129)I in corals from the southern hemisphere show the increasing trend of (129)I concentrations in the marine environment caused by HNA. Here we show time series of (129)I/(127(stable))I isotopic ratios in two coral cores from the northern hemisphere (Philippines) and how these record (129)I released from HNA in even greater, unprecedented detail. Corals were taken from the Pacific Ocean (Baler) and South China Sea (Parola) sides of the Philippines. We observed nearly identical peaks ((129)I/(127)I ∼ 31.5 × 10(-12)) in both the Baler and Parola records, each attributed to the year 1962 - the year with the highest recorded amount of (129)I release from nuclear bomb testing. This 1962 (129)I bomb signal offers a new time marker that can be used to establish or confirm age models of corals, comparable to or possibly better than the well-known coral (14)C bomb peak. We also observed nuclear fuel reprocessing and Chernobyl accident (129)I signals in years 1977, 1980, and 1986, concurrently in Parola and with 9 to 11-year lags in Baler. This discrepancy in timing suggests that (129)I was transported to the South China Sea and Pacific Ocean sides of the Philippines directly from the atmosphere and through prevailing ocean currents, respectively. Lastly, we observed surprisingly high (129)I/(127)I isotopic ratios (i.e., 22.8 to 38.9 × 10(-12)) in the Parola record after the year 1996, which is in contrast to the decreasing trend observed in the Baler record and in published (129)I releases of different HNA. These results possibly indicate the presence of unknown sources of (129)I in the South China Sea region.

  7. Sinuosity change of the Po River near Cremona (Northern Italy) - a result of neotectonic activity?

    NASA Astrophysics Data System (ADS)

    Petrovszki, Judit; Timár, Gábor

    2010-05-01

    In the map sheets of the Second Military Survey of the Habsburg Empire, Lombardia, Parma, Modena and Venice also can be seen (Timár et al., 2006). This area was surveyed between 1818 and 1829. In these map sheets, we can also follow the river Po from Vaccarizza to the delta. This river reach is about 350 km long. This river reach was digitized and sinuosity values were calculated with different window sizes, and displayed in a spectrum-like diagram (sinuosity spectra; after van Balen et al., 2008). At Cremona, a significante sinuosity change were identified. The sinuosity increasing, and we have high sinuosity values. In the summarizing geological map of Italy (Compagnoni and Calluzzo, 2004), at this place, a tectonic line was identified. So probably this fault line invokes the sinuosity change on the river. The vertical movements indicated on the maps are just the opposite like they would be according to the flume experiments of Ouchi (1985). In the case of the Po River at Cremona, the decrease of the channel slope results higher sinuosity. The reason is that the rate of the slope and water discharge is higher than it is required by the self-organized meandering and the river parameters fell to the range of the unorganized meandering (cf. Timár, 2003). Another possible explanation could be that the northern tributary, the Adda River has significant sediment load that lowers the sinuosity of the trunk river at the confluence. Compagnoni, B., Galluzzo, F. (eds., 2004): Geological Map of Italy. Agenzia per la Protezione dell'Ambiente per I Servizi Tecnici - Dipartimento Difesa del Suolo, Servizio Geologico d'Italia, Rome-Florence-Genoa. Map, scale=1:1250000, especially printed for the 32nd International Geological Congress. Ouchi, S. (1985): Response of alluvial rivers to slow active tectonic movement. Geol. Soc. Am. Bull. 96: 504-515. Timár, G. (2003): Controls on channel sinuosity changes: a case study of the Tisza River, the Great Hungarian Plain. Quaternary

  8. Northern polar dunes: a spring activity analog to that seen in southern polar terrains

    NASA Astrophysics Data System (ADS)

    Portyankina, Ganna; Thomas, Nicolas; Pommerol, Antoine; Aye, Klaus-Michael

    The High Resolution Imaging Science Experiment (HiRISE) on the Mars Reconnaissance Or-biter (MRO) is presently (February 2010) observing its second northern spring on Mars. Com-plimentary spectral information from CRISM is available to gain insight into the composition and physical state of the surface. In the north polar areas, there are active regions. Here, dark dunes extend almost circumferen-tially around the permanent polar cap. This area exhibits phenomena similar to those observed in south polar areas in southern spring. Among these phenomena are cracks in a translucent ice layer which are considered to be manifestations of ice damage by increasing pressure at the interface between the ice layer and substrate underlying it [1]. Bright and dark fans, attributed to cold jet activity, are also observed on top of dunes [2]. The presence of these features indi-cates that dunes are covered by a conformal ice layer and that this layer is at least partially transparent to solar radiation. CO2 ice sublimation and processes connected to it appear to produce a variety of observable features. Yet another distinctive feature observed in this area (and rarely present in the south) is the presence of dark slope streaks on dunes. They originate from the crests of dunes and run down following the gravitational potential. They lengthen as the season progresses. Their origin has been discussed to be possible liquid brine flows [3, 4]. In the present work we report on indications that these slope streaks are instead dry mass wasting process. Due to geometry of solar illumination at local latitude the energy input on the crest is higher than in the valley between dunes. The ice cover thins most quickly in areas closest to the crest. In addition, applying stress distribution calculations to the conformal ice layer on realistic dune geometry, we show that the crest of the dune is the weakest point for the ice bending stress. The rupture of the ice layer therefore happens close to

  9. 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

  10. Seasonally different response of photosynthetic activity to daytime and night-time warming in the Northern Hemisphere

    SciTech Connect

    Tan, Jianguang; Piao, Shilong; Chen, Anping; Zeng, Zhenzhong; Ciais, Philippe; Janssens, Ivan A.; Mao, Jiafu; Myneni, Ranga B.; Peng, Shushi; Peñuelas, Josep; Shi, Xiaoying; Vicca, Sara

    2014-08-27

    Over the last century the Northern Hemisphere has experienced rapid climate warming, but this warming has not been evenly distributed seasonally, as well as diurnally. The implications of such seasonal and diurnal heterogeneous warming on regional and global vegetation photosynthetic activity, however, are still poorly understood. Here, we investigated for different seasons how photosynthetic activity of vegetation correlates with changes in seasonal daytime and night-time temperature across the Northern Hemisphere (>30°N), using Normalized Difference Vegetation Index (NDVI) data from 1982 to 2011 obtained from the Advanced Very High Resolution Radiometer (AVHRR). Our analysis revealed some striking seasonal differences in the response of NDVI to changes in day- versus night-time temperatures. For instance, while higher daytime temperature (Tmax) is generally associated with higher NDVI values across the boreal zone, the area exhibiting a statistically significant positive correlation between Tmax and NDVI is much larger in spring (41% of area in boreal zone – total area 12.6 × 106 km2) than in summer and autumn (14% and 9%, respectively). In contrast to the predominantly positive response of boreal ecosystems to changes in Tmax, increases in Tmax tended to negatively influence vegetation growth in temperate dry regions, particularly during summer. Changes in night-time temperature (Tmin) correlated negatively with autumnal NDVI in most of the Northern Hemisphere, but had a positive effect on spring and summer NDVI in most temperate regions (e.g., Central North America and Central Asia). Such divergent covariance between the photosynthetic activity of Northern Hemispheric vegetation and day- and night-time temperature changes among different seasons and climate zones suggests a changing dominance of ecophysiological processes across time and space. Lastly, understanding the seasonally

  11. 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

  12. Weather and Large-Scale Dust Activity during Martian Northern Spring and Summer

    NASA Astrophysics Data System (ADS)

    Kass, David M.; Kleinboehl, Armin; McCleese, Daniel J.; Schofield, John Tim; Smith, Michael D.; Heavens, Nicholas

    2016-10-01

    Observations from MCS, TES and THEMIS now span the northern spring and summer seasons (Ls 0° to 180°) of 10 consecutive Mars Years (MY 24 through MY 33). These observations show very similar behavior each year. However, there are also noticeable differences and clear signs of inter-annual variability. To best study the three datasets, we examine zonal mean observations of the lower atmosphere (50 Pa, or ~25 km). This region was selected to provide the best quality from all three instruments. We separate the daytime (afternoon) and nighttime (early morning) data in the analysis.The climate at these seasons is dominated by the aphelion cloud belt, and 50 Pa is often close to the peak opacities in the clouds. There is also a strong diurnal thermal tide signature throughout the season at this altitude. The overall behavior is a rapid cooling at the start of the year (as the dust from the dusty season sediments out of the atmosphere) over the the first ~30° of Ls. The coldest temperatures then last until about the solstice and are followed by a slow warming trend through most of the rest of the season. The last ~30° prior to the fall equinox show a more rapid warming trend and significant inter-annual variability. In about half of the years, there is a warming event of the 50 Pa temperatures in the second half of northern summer. The warming is the signature of dust being lofted above the boundary layer, into the lower atmosphere. Due to the relatively clear atmosphere overall, even modest amounts of dust will create noticeable temperature changes. The temperature signature of the dust is more pronounced in the northern hemisphere.

  13. Lack of Active Onchocerca volvulus Transmission in the Northern Chiapas Focus of Mexico

    PubMed Central

    Rodríguez-Pérez, Mario A.; Unnasch, Thomas R.; Domínguez-Vázquez, Alfredo; Morales-Castro, Alba L.; Richards, Frank; Peña-Flores, Graciela P.; Orozco-Algarra, María Eugenia; Prado-Velasco, Gibert

    2010-01-01

    The northern Chiapas onchocerciasis focus has undergone 11 years of ivermectin mass treatment. No evidence of microfilariae in the cornea and/or anterior chamber of the eye or in skin snips was seen in residents examined in 2006 in two sentinel communities (upper limit of the 95% confidence interval [UL 95% CI] = 0.5% and 0.3%, respectively). In children 10 and under, 0 of 305 were found to harbor antibodies to Ov16, a marker of parasite exposure; 0 of 4,400 Simulium ochraceum s.l. collected in 2005 contained parasite DNA, giving an UL 95% CI for the infective rate of 0.9/2,000, and an UL 95% CI of the seasonal transmission potential of 1.2 L3/person. These data, assumed to be representative of the focus as a whole, suggest that there is no ongoing transmission of Onchocerca volvulus in the northern Chiapas focus. Community-wide treatments with ivermectin were halted in 2008, and a post-treatment surveillance phase was initiated. PMID:20595471

  14. 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.

  15. 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%.

  16. Assessment of active faults for maximum credible earthquakes of the southern California-northern Baja region

    NASA Astrophysics Data System (ADS)

    Slemmons, D. B.; Omalley, P.; Whitney, R. A.; Chung, D. H.; Bernreuter, D. L.

    1982-06-01

    Compilation of a data base is presented for maximum or maximum credible earthquakes that can be used to compute seismic hazard spectra at the San Onofre Nuclear Generating Stations in southern California. Estimates of fault slip rate and estimated recurrence - northern Baja California region are given. According to a direct relationship between the total fault length and the earthquake magnitude, the maximum earthquake for the Offshore Zone of Deformation (OZD) is estimated to be of about 6.8 or 6.9 surface wave magnitude. Another empirical relationship relating the fractional fault length and earthquake magnitude for strike slip faults results in an estimated maximum earthquake of about M/sub S/ = 6.8 for the OZD.

  17. Relationship Between Ecosystem Productivity and Photosynthetically Active Radiation for Northern Peatlands

    NASA Technical Reports Server (NTRS)

    Frolking, S. E.; Bubier, J. L.; Moore, T. R.; Ball, T.; Bellisario, L. M.; Bhardwaj, A.; Carroll, P.; Crill, P. M.; Lafleur, P. M.; McCaughey, J. H.; Roulet, N. T.; Suyker, A. E.; Verma, S. B.; Waddington, J. M.; Whiting, G. J.

    1998-01-01

    We analyzed the relationship between net ecosystem exchange of carbon dioxide (NEE) and irradiance (as photosynthetic photon flux density or PPFD), using published and unpublished data that have been collected during midgrowing season for carbon balance studies at seven peatlands in North America and Europe, NEE measurements included both eddy-correlation tower and clear, static chamber methods, which gave very similar results. Data were analyzed by site, as aggregated data sets by peatland type (bog, poor fen, rich fen, and all fens) and as a single aggregated data set for all peatlands. In all cases, a fit with a rectangular hyperbola (NEE = alpha PPFD P(sub max)/(alpha PPFD + P(sub max) + R) better described the NEE-PPFD relationship than did a linear fit (NEE = beta PPFD + R). Poor and rich fens generally had similar NEE-PPFD relationships, while bogs had lower respiration rates (R = -2.0 micro mol m(exp -2) s(exp -1) for bogs and -2.7 micro mol m(exp -2) s(exp -1)) for fens) and lower NEE at moderate and high light levels (P(sub max)= 5.2 micro mol m(exp -2) s(exp -1) for bogs and 10.8 micro mol m(exp -2) s(exp -1) for fens). As a single class, northern peatlands had much smaller ecosystem respiration (R = -2.4 micro mol m(exp -2) s(exp -1)) and NEE rates (alpha = 0.020 and P(sub max)= 9.2 micro mol m(exp -2) s(exp -1)) than the upland ecosystems (closed canopy forest, grassland, and cropland). Despite this low productivity, northern peatland soil carbon pools are generally 5-50 times larger than upland ecosystems because of slow rates of decomposition caused by litter quality and anaerobic, cold soils.

  18. Structure of the active rift zone and margins of the northern Imperial Valley from Salton Seismic Imaging Project (SSIP) data

    NASA Astrophysics Data System (ADS)

    Livers, A.; Han, L.; Delph, J. R.; White-Gaynor, A. L.; Petit, R.; Hole, J. A.; Stock, J. M.; Fuis, G. S.

    2012-12-01

    First-arrival refraction data were used to create a seismic velocity model of the upper crust across the actively rifting northern Imperial Valley and its margins. The densely sampled seismic refraction data were acquired by the Salton Seismic Imaging Project (SSIP) , which is investigating rift processes in the northern-most rift segment of the Gulf of California extensional province and earthquake hazards at the southern end of the San Andreas Fault system. A 95-km long seismic line was acquired across the northern Imperial Valley, through the Salton Sea geothermal field, parallel to the five Salton Butte volcanoes and perpendicular to the Brawley Seismic Zone and major strike-slip faults. Nineteen explosive shots were recorded with 100 m seismometer spacing across the valley and with 300-500 m spacing into the adjacent ranges. First-arrival travel times were picked from shot gathers along this line and a seismic velocity model was produced using tomographic inversion. Sedimentary basement and seismic basement in the valley are interpreted to be sediment metamorphosed by the very high heat flow. The velocity model shows that this basement to the west of the Brawley Seismic Zone is at ~4-km depth. The basement shallows to ~2-km depth in the active geothermal field and Salton Buttes volcanic field which locally coincide with the Brawley Seismic Zone. At the eastern edge of the geothermal field, the basement drops off again to ~3.5-km depth. The eastern edge of the valley appears to be fault bounded by the along-strike extension of the Sand Hills Fault, an inactive strike-slip fault. The seismic velocities to the east of the fault correspond to metamorphic rock of the Chocolate Mountains, different from the metamorphosed basement in the valley. The western edge of the valley appears to be fault bounded by the active Superstition Hills Fault. To the west of the valley, >4-km deep valley basement extends to the active Superstition Hills Fault. Basement then shallows

  19. Barriers and Facilitators to Being Physically Active on a Rural U.S. Northern Plains American Indian Reservation

    PubMed Central

    Jahns, Lisa; McDonald, Leander R.; Wadsworth, Ann; Morin, Charles; Liu, Yan

    2014-01-01

    The objective of the present study was to identify barriers to and facilitators of physical activity among American Indian adults living on a rural, U.S. Northern Plains reservation using the nominal group technique (NGT). NGT is a method of data generation and interpretation that combines aspects of qualitative (free generation of responses) and quantitative (systematic ranking of responses) methodologies. Adults participated in one of two NGT sessions asking about either barriers to (n = 6), or facilitators of (n = 5), being physically active. Participants nominated and ranked 21 barriers and 18 facilitators. Barriers indicated lack of knowledge of how to fit physical activity into a daily schedule, work, caring for family members, and prioritizing sedentary pursuits. Other responses included environmental barriers such as lack of access and transportation to a gym, unsafe walking conditions, and inclement weather. Facilitators to following recommendations included knowledge of health benefits of physical activity and the perception of physical activity as enjoyable, including feeling good when working out. Environmental facilitators included being outdoors walking and biking as well as parks and exercise facilities. Responses provided direction for locally designed community-based programs to promote facilitators and decrease barriers to individual’s engagement in physical activity. PMID:25421064

  20. Plasma Plasminogen Activator Inhibitor-1 Is Associated with End-Stage Proliferative Diabetic Retinopathy in the Northern Chinese Han Population

    PubMed Central

    Zhong, Ze-Long; Chen, Song

    2012-01-01

    Objective. To identify predictors of end-stage proliferative diabetic retinopathy (PDR) in a cohort of individuals with type 2 diabetes mellitus (T2DM) from the Northern Chinese Han population. Methods. We investigated characteristics of 153 consecutive diabetic patients with end-stage PDR (62 males, 91 females), 123 consecutive PDR patients without end-stage PDR (48 males, 75 females), and 151 normal subjects (63 males, 88 females). Only one eye of each patient or healthy subject was included in this study. Univariate logistic regression models and multivariate logistic regression models were constructed to evaluate the predictors of end-stage PDR. Results. In univariate analysis, systolic blood pressure, diastolic blood pressure, duration of diabetes, family history of T2DM, and plasminogen activator inhibitor-1 (PAI-1) were significently associated with end-stage PDR. After multivariate analysis, family history of T2DM, plasma PAI-1 levels, smoking, and duration of diabetes were four positive predictors associated with end-stage PDR. Conclusions. Higher plasma levels of PAI-1 were associated with end-stage PDR in the Northern Chinese Han population with T2DM. PMID:23304115

  1. The upper Pleistocene on the northern face of the Guadarrama Mountains (central Spain): Palaeoclimatic phases and glacial activity

    NASA Astrophysics Data System (ADS)

    Bullón, Teresa

    2016-09-01

    The present paper provides new information on Pleistocene glacial activity in a mountainous area of the Iberian Central System. A sediment analysis associated with Pleistocene modelling was carried out using: (1) granulometric and morphometric procedures, (2) quartz grain microtexture techniques (SEM) to discriminate between glacial and no glacial origins of sediments, (3) clay X-ray diffraction study to determine intra-Pleistocene climate variability, and (4) optically stimulated luminescence (OSL) absolute dating. The results show that the sediments were formed in two different phases associated with glacial dynamics, one of them was 35-30 ky BP and another was 25-20 ky BP, separated by a short intermediate warm-wet period. Identification of glacial phenomena is new for the northern slopes of the Guadarrama Mountains (facing the north Meseta, Duero basin), although they are not unusual within the general context of the Iberian Central System. From the data provided, we deduce that glaciation in these mountains was much more intense and widespread than had previously been thought because, on the northern slopes, glaciers occupied large areas reaching the base of the mountains. The evidence favours new interpretations of Pleistocene morphology in the centre of the Iberian Peninsula and, by extension, on the southwestern edge of Europe; it also highlights the sensitivity of mountainous areas with regard to Quaternary climate changes.

  2. Northern Australia

    Atmospheric Science Data Center

    2013-04-16

    article title:  Tropical Northern Australia     View Larger Image ... Imaging SpectroRadiometer (MISR) images of tropical northern Australia were acquired on June 1, 2000 (Terra orbit 2413) during the long dry ...

  3. Contribution of geophysical data in delineating the active subsurface structures along the southeastern Mediterranean and northern Egypt

    NASA Astrophysics Data System (ADS)

    Saleh, Salah; Elwan, Mostafa

    2016-04-01

    The solution obtained with 3D Euler deconvolution gives better-focused depth estimates, which are closer to the real position of sources; the results presented here can be used to constrain depth to active crustal structures (fault system, magmatic activity and subduction zones) for southeastern Mediterranean and northern Egypt. The results indicated that the area was affected by sets of structural systems, which primarily trended in the NE-SW to NNE-SSW and NW-SE, directions. The estimated shallow Bouguer results (for SI dikes, sills and flows models) shows an abrupt change depth values (1-8 km) almost clustered along the eastern segment of the Cyprian arc, Indicates continuation of the ophiolite at a depth below younger sediments of the Latakia and southern Antalya Basins. This means that, the Eratosthenes Seamount block is in the process of dynamically subsiding beneath Cyprus to the north and thrusted onto the Levantine Basin to the south. Nevertheless, the Cyprian arc region is dominated by several compressional shallow fault systems (0-4 km) trended E-W to WNW direction, which may initiated due to the northward-directed movement of Africa and ongoing subducting the Levant oceanic segment beneath the Tauric arc south of Cyprus. However, the deep magnetic depths (28 km, for SI of dipole model) correspond more closely to the Moho depth in the oceanic regions of marine part, with a high proportion of Younger Granites (Eratosthenes Seamount, Florence Rise, South Cyprus, eastern part of Levantine Basin and north Nile Cone) which are accompanied with low heat flow values. We can state that the clustering of most shallow earthquakes along this structures (especially beneath southern part of Florence Rise) may most likely be attributed to the active mantle upwelling (volcanic earthquakes), which are ultimately related to volcanic processes. Whereas, the Moho depth reaches to 35 km beneath the continental regions of Nile Delta basins and northern Sinai, which are

  4. Using the Landsat Thematic Mapper to detect and monitor active volcanoes - An example from Lascar volcano, northern Chile

    NASA Technical Reports Server (NTRS)

    Francis, P. W.; Rothery, D. A.

    1987-01-01

    The Landsat Thematic Mapper (TM) offers a means of detecting and monitoring thermal features of active volcanoes. Using the TM, a prominent thermal anomaly has been discovered on Lascar volcano, northern Chile. Data from two short-wavelength infrared channels of the TM show that material within a 300-m-diameter pit crater was at a temperature of at least 380 C on two dates in 1985. The thermal anomaly closely resembles in size and radiant temperature the anomaly over the active lava lake at Erta'ale in Ethiopia. An eruption took place at Lascar on Sept. 16, 1986. TM data acquired on Oct. 27, 1986, revealed significant changes within the crater area. Lascar is in a much more active state than any other volcano in the central Andes, and for this reason it merits further careful monitoring. Studies show that the TM is capable of confidently identifying thermal anomalies less than 100 m in size, at temperatures of above 150 C, and thus it offers a valuable means of monitoring the conditions of active or potentially active volcanoes, particularly those in remote regions.

  5. Seasonal Variation in Physical Activity among Preschool Children in a Northern Canadian City

    ERIC Educational Resources Information Center

    Carson, Valerie; Spence, John C.; Cutumisu, Nicoleta; Boule, Normand; Edwards, Joy

    2010-01-01

    Little research has examined seasonal differences in physical activity (PA) levels among children. Proxy reports of PA were completed by 1,715 parents on their children in Edmonton, Alberta, Canada. Total PA (TPA) minutes were calculated, and each participant was classified as active, somewhat active, or inactive. Logistic regression models were…

  6. Simulation of water resource and its relation to urban activity in Dalian City, Northern China

    NASA Astrophysics Data System (ADS)

    Nakayama, T.; Sun, Y.; Geng, Y.

    2010-09-01

    The NIES Integrated Catchment-based Eco-hydrology (NICE) model was applied to the Biliu River catchment, northern China, to estimate the carrying capacity of the water resource there. The model reproduced well the water and heat budgets after the construction of a reservoir in the middle reach of the river. It also correctly backcasted the degradation of water resources such as river discharge and groundwater after the completion of the reservoir. Calculation of the normalized difference vegetation index (NDVI) from satellite data clearly showed vegetation degradation downstream of the reservoir. Statistical analysis of a decoupling indicator based on the simulated water carrying capacity and on the satellite data of vegetation index indicated that water-related stress in Dalian city, where the economy has grown rapidly after the completement of the reservoir, has increased in accordance with the environmental degradation below the reservoir. The results indicate a close relationship between water resource and economic growth, which has greatly affected ecosystem degradation and its serious burden on the environment in the catchment. The simulated results highlight the linkage between urban development in Dalian and sustainable water resource management.

  7. Studies of the Northern Alaskan Coastal System: Ongoing project work and synthesis activities

    NASA Astrophysics Data System (ADS)

    Douglas, T. A.; Sturm, M.; Ashjian, C. J.; Jorgensen, T.; Oechel, W. C.; Ping, C.; Rhew, R. C.; Stieglitz, M.

    2006-12-01

    Six ongoing projects focus on a better understanding of processes occurring along the Arctic Alaskan Coast. These projects, grouped as "Studies of the Northern Alaskan Coastal System", or SNACS, combine field, laboratory, modeling and human dimensions research. They include: 1) an investigation of climate variability, ocean processes, sea ice, bowhead whales, and Inupiat subsistence whaling, 2) research on the impact of variability within the ocean and atmosphere on terrestrial fluxes of carbon dioxide, dissolved organic matter and energy, 3) an inventory and description of soil organic carbon fluxes and ground ice in the coastal environment, 4) a determination of whether arctic coastal terrestrial ecosystems are significant sources or sinks of atmospheric methyl halides, chloroform and methane, 5) development of generalized discharge- constituent relationships for arctic basins, and 6) an investigation of the processes controlling mercury deposition to the coastal system. Three broad themes unite the projects: 1) nutrient fluxes from rivers and shoreline erosion in the Arctic coastal zone, 2) impacts of cryospheric changes on the Alaskan Arctic Coast, and 3) potential rapid regime shifts controlled by atmospheric and meteorological processes that could affect the Alaskan Arctic Coast. Warming of the Arctic, particularly its impact on sea ice and nutrient transport in arctic rivers is already affecting fundamental coastal system processes. The six SNACS projects are helping to understand how these impacts will evolve and what their ramifications will be both within and outside of the Arctic.

  8. Evidence for a weakening relationship between interannual temperature variability and northern vegetation activity.

    PubMed

    Piao, Shilong; Nan, Huijuan; Huntingford, Chris; Ciais, Philippe; Friedlingstein, Pierre; Sitch, Stephen; Peng, Shushi; Ahlström, Anders; Canadell, Josep G; Cong, Nan; Levis, Sam; Levy, Peter E; Liu, Lingli; Lomas, Mark R; Mao, Jiafu; Myneni, Ranga B; Peylin, Philippe; Poulter, Ben; Shi, Xiaoying; Yin, Guodong; Viovy, Nicolas; Wang, Tao; Wang, Xuhui; Zaehle, Soenke; Zeng, Ning; Zeng, Zhenzhong; Chen, Anping

    2014-10-16

    Satellite-derived Normalized Difference Vegetation Index (NDVI), a proxy of vegetation productivity, is known to be correlated with temperature in northern ecosystems. This relationship, however, may change over time following alternations in other environmental factors. Here we show that above 30°N, the strength of the relationship between the interannual variability of growing season NDVI and temperature (partial correlation coefficient RNDVI-GT) declined substantially between 1982 and 2011. This decrease in RNDVI-GT is mainly observed in temperate and arctic ecosystems, and is also partly reproduced by process-based ecosystem model results. In the temperate ecosystem, the decrease in RNDVI-GT coincides with an increase in drought. In the arctic ecosystem, it may be related to a nonlinear response of photosynthesis to temperature, increase of hot extreme days and shrub expansion over grass-dominated tundra. Our results caution the use of results from interannual time scales to constrain the decadal response of plants to ongoing warming.

  9. Coastal Marine Terraces Define Late Quaternary Fault Activity and Deformation Within Northern East Bay Hills, San Francisco Bay Region

    NASA Astrophysics Data System (ADS)

    Kelson, K. I.

    2004-12-01

    Detailed mapping of uplifted marine platforms bordering the Carquinez Strait between Benicia and Pinole, California, provides data on the pattern and rate of late Quaternary deformation across the northern East Bay Hills. Field mapping, interpretation of early 20th-century topographic data, analysis of aerial photography, and compilation of onshore borehole data show the presence of remnants of three platforms, with back-edge elevations of about 4 m, 12 m, and 18 m. Based on U-series dates (Helley et al., 1993) and comparison of platform elevations to published sea-level curves, the 12-m-high and 18-m-high platforms correlate with substage 5e (ca. 120 ka) and stage 9 (ca. 330 ka) sea-level high stands, respectively. West of the Southhampton fault, longitudinal profiles of platform back-edges suggest that the East Bay Hills between Pinole and Vallejo have undergone block uplift at a rate of 0.05 +/- 0.01 m/ka without substantial tilting or warping. With uncertainty of <3 m, the 120 ka and 330 ka platforms are at the same elevations across the NW-striking Franklin fault. This west-vergent reverse fault previously was interpreted to have had late Pleistocene activity and to accommodate crustal shortening in the East Bay Hills. Our data indicate an absence of vertical displacement across the Franklin fault within at least the past 120ka and perhaps 330ka. In contrast, the stage 5e and 9 have up-on-the-east vertical displacement and gentle westward tilting across the N-striking Southhampton fault, with a late Pleistocene vertical slip rate of >0.02 m/ka. The northerly strike and prominent geomorphic expression of this potentially active fault differs from the Franklin fault. Our mapping of the Southhampton fault suggests that it accommodates dextral shear in the East Bay Hills, and is one of several left-stepping, en echelon N-striking faults (collectively, the "Contra Costa shear zone", CCSZ) in the East Bay Hills. Faults within this zone coincide with geomorphic

  10. Timely detection of localized excess influenza activity in Northern California across patient care, prescription, and laboratory data.

    PubMed

    Greene, Sharon K; Kulldorff, Martin; Huang, Jie; Brand, Richard J; Kleinman, Kenneth P; Hsu, John; Platt, Richard

    2011-02-28

    Timely detection of clusters of localized influenza activity in excess of background seasonal levels could improve situational awareness for public health officials and health systems. However, no single data type may capture influenza activity with optimal sensitivity, specificity, and timeliness, and it is unknown which data types could be most useful for surveillance. We compared the performance of 10 types of electronic clinical data for timely detection of influenza clusters throughout the 2007/08 influenza season in northern California. Kaiser Permanente Northern California generated zip code-specific daily episode counts for: influenza-like illness (ILI) diagnoses in ambulatory care (AC) and emergency departments (ED), both with and without regard to fever; hospital admissions and discharges for pneumonia and influenza; antiviral drugs dispensed (Rx); influenza laboratory tests ordered (Tests); and tests positive for influenza type A (FluA) and type B (FluB). Four credible events of localized excess illness were identified. Prospective surveillance was mimicked within each data stream using a space-time permutation scan statistic, analyzing only data available as of each day, to evaluate the ability and timeliness to detect the credible events. AC without fever and Tests signaled during all four events and, along with Rx, had the most timely signals. FluA had less timely signals. ED, hospitalizations, and FluB did not signal reliably. When fever was included in the ILI definition, signals were either delayed or missed. Although limited to one health plan, location, and year, these results can inform the choice of data streams for public health surveillance of influenza.

  11. Consistent QBO-dependent effect of geomagnetic activity on the Northern Annular Mode during the 20th century

    NASA Astrophysics Data System (ADS)

    Maliniemi, Ville; Asikainen, Timo; Mursula, Kalevi

    2016-04-01

    Several earlier studies have shown that geomagnetic activity (GA), as a proxy for energetic particle precipitation into the atmosphere, affects the winter-time Northern Annular Mode (NAM), which is the dominant circulation pattern in the northern hemisphere during winter. It has also been found that the quasi-biennial oscillation (QBO) modulates the relationship between GA and NAM. However, some of the earlier studies on this QBO modulation have been mutually conflicting, with some studies suggesting a stronger positive relation in the easterly phase of the QBO, while other studies suggest a stronger positive relation in the westerly phase of the QBO. Here we study the QBO-GA-NAM relationship using a QBO reconstruction covering the whole 20th century. We find that the QBO modulation of the GA-NAM relation is temporally variable, which explains the earlier, seemingly differing results. Positive GA-NAM relation is found to be valid in the easterly QBO phase at 30 hPa during the whole 20th century. We also find that the QBO at 30 hPa represents the Holton-Tan relation for the surface circulation better than QBO at 50 hPa, and that the Holton-Tan relation is only observed during early/mid winter, while an anti-Holton-Tan relation is found in the late winter for strong geomagnetic activity. These results emphasize the variable but systematic response of NAM to energetic particle precipitation during the entire 20th century, and underline the importance of considering the preconditioning of the atmosphere when studying the solar-related effects upon climate.

  12. Unusual Polar Activity of the Sun in the Northern Hemisphere and Its Implications for Solar Cycle 25

    NASA Astrophysics Data System (ADS)

    Gopalswamy, Nat; Masuda, Satoshi; Yashiro, Seiji; Akiyama, Sachiko; Shibasaki, Kiyoto

    2016-07-01

    Polar field strength in one solar cycle is known to indicate the strength (e.g., Sunspot number) and phase of the next cycle. In particular the polar field strength (or its proxies such as the polar coronal hole area and microwave polar brightness) during the minimum phase of a given cycle seem to be well correlated with the maximum sunspot number of the next cycle. Polar prominence eruptions and coronal mass ejections have also been found to be indicators of low polar field; their cessation signals the time of polarity reversal. While these indicators are present in the current cycle, significant differences are found regarding the phase lag between the two hemispheres and the duration of polar eruptions. We use data from the Nobeyama Radioheliograph, the Solar Dynamics Observatory, SOLIS, and Wilcox Solar Observatory to highlight these differences. We find that the north polar region of the Sun has near-zero field strength for more than three years. This is unusually long and caused by surges of both polarities heading toward the north pole that prevent the buildup of the polar field. This seems to be due to anti-Hale active regions that appeared around the 2012 peak sunspot activity in the northern hemisphere. The unusual condition is consistent with (i) the continued high-latitude prominence eruption, (ii) the extended period of high tilt angle of the heliospheric current sheet, (iii) the weak microwave polar brightness, and (iv) the lack of north polar coronal hole. On the other hand, the south polar field has started building up and the coronal hole has appeared in early 2015 because of large active regions of the correct tilt in the southern hemisphere during the 2014 peak of sunspot activity. The extended period of near-zero field in the north polar region should result in very weak and delayed sunspot activity in the northern hemisphere in cycle 25. On the other hand the south polar field has already increased significantly, suggesting that the activity in

  13. Muskox Milk, Ptarmigan Pie and Other Northern Delicacies (Kids Can Cook): Recipes and Related Activities.

    ERIC Educational Resources Information Center

    Northwest Territories Dept. of Education, Yellowknife.

    This profusely illustrated book of recipes encourages the use of foods in children's learning activities, especially foods that are harvested in the Northwest Territories of Canada and that play a role in health maintenance. The activities described also provide an opportunity for children to experience new foods. Introductory material includes…

  14. The Status of Daily Physical Activity in Northern Ontario's Elementary Public Schools

    ERIC Educational Resources Information Center

    Rickwood, Greg

    2015-01-01

    The importance of daily physical activity (DPA) for children and youth is highlighted in Canada's Active Healthy Kids annual report on fitness. Since 2005, this report has revealed that elementary-aged students are spending most of their waking hours stationary, in front of screens (phones, televisions, computers). To counteract this trend, the…

  15. Relationship between persistent halogenated organic contaminants and TCDD-toxic equivalents on EROD activity and retinoid and thyroid hormone status in northern fulmars.

    PubMed

    Helgason, Lisa B; Verreault, Jonathan; Braune, Birgit M; Borgå, Katrine; Primicerio, Raul; Jenssen, Bjørn M; Gabrielsen, Geir W

    2010-11-15

    We investigated whether the hepatic cytochrome P450 1A activity (measured as 7-ethoxyresorufin-O-deethylase (EROD)) and plasma thyroid hormone and liver retinoid concentrations were explained by liver and blood levels of halogenated organic contaminants (HOCs) in free-ranging breeding northern fulmars (Fulmarus glacialis) from Bjørnøya in the Norwegian Arctic. Hepatic EROD activity and liver levels of 2,3,7,8-tetrachlorodibenzo-p-dioxin toxic equivalents (TEQs) were positively correlated, suggesting that hepatic EROD activity is a good indicator for dioxin and dioxin-like HOC exposure in breeding northern fulmars. There were not found other strong relationships between HOC concentrations and hepatic EROD activity, plasma thyroid or liver retinoid concentrations in the breeding northern fulmars. It is suggested that the HOC levels found in the breeding northern fulmars sampled on Bjørnøya were too low to affect plasma concentrations of thyroid hormones and liver levels of retinol and retinyl palmitate, and that hepatic EROD activity is a poor indicator of polychlorinated biphenyl (PCB) and pesticide exposure.

  16. Evaluation of the flora of northern Mexico for in vitro antimicrobial and antituberculosis activity.

    PubMed

    Molina-Salinas, G M; Pérez-López, A; Becerril-Montes, P; Salazar-Aranda, R; Said-Fernández, S; de Torres, N Waksman

    2007-02-12

    The aim of the present study was to evaluate the potential antimicrobial activity of 14 plants used in northeast México for the treatment of respiratory diseases, against drug-sensitive and drug-resistant strains of Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae type b and Mycobacterium tuberculosis. Forty-eight organic and aqueous extracts were tested against these bacterial strains using a broth microdilution test. No aqueous extracts showed antimicrobial activity, whereas most of the organic extracts presented antimicrobial activity against at least one of the drug-resistant microorganisms tested. Methanol-based extracts from the roots and leaves of Leucophyllum frutescens and ethyl ether extract from the roots of Chrysanctinia mexicana showed the greatest antimicrobial activity against the drug-resistant strain of Mycobacterium tuberculosis; the minimal inhibitory concentration (MIC) were 62.5, 125 and 62.5 microg/mL, respectively; methanol-based extract from the leaves of Cordia boissieri showed the best antimicrobial activity against the drug-resistant strain of Staphylococcus aureus (MIC 250 microg/mL); the hexane-based extract from the fruits of Schinus molle showed considerable antimicrobial activity against the drug-resistant strain of Streptococcus pneumoniae (MIC 62.5 microg/mL). This study supports that selecting plants by ethnobotanical criteria enhances the possibility of finding species with activity against resistant microorganisms.

  17. Active tectonics in northern Victoria Land (Antarctica) inferred from the integration of GPS data and geologic setting

    NASA Astrophysics Data System (ADS)

    Dubbini, M.; Cianfarra, P.; Casula, G.; Capra, A.; Salvini, F.

    2010-12-01

    A semipermanent Global Positioning System (GPS) network of 30 vertices known as the Victoria Land Network for Deformation Control (VLNDEF) was set up in the Austral summer of 1998 in northern Victoria Land (NVL), including Terra Nova Bay (TNB), Antarctica. The locations were selected according to the known Cenozoic fault framework, which is characterized by a system of NW-SE regional faults with right-lateral, strike-slip kinematics. The TNB1 permanent GPS station is within the VLNDEF, and following its installation on a bedrock monument in October 1998, it has been recording almost continuously. The GPS network has been surveyed routinely every two summers, using high-quality, dual-frequency GPS receivers. In this study we present the results of a distributed session approach applied to the processing of the GPS data of the VLNDEF. An improved reference frame definition was implemented, including a new Euler pole, to compute the Antarctic intraplate residual velocities. The projection of the residual velocities on the main faults in NVL show present-day activities for some faults, including the Tucker, Leap Year, Lanterman, Aviator, and David faults, with right-lateral strike-slip kinematics and local extensional and compressional components. This active fault pattern divides NVL into eight rigid blocks, each characterized by its relative movements and rigid rotations. These show velocities of up to several millimeters per year, which are comparable to those predicted by plate tectonic theory at active plate margins.

  18. Phenolic substance characterization and chemical and cell-based antioxidant activities of 11 lentils grown in the northern United States.

    PubMed

    Xu, Baojun; Chang, Sam K C

    2010-02-10

    Chemical and cellular antioxidant activities and phenolic profiles of 11 lentil cultivars grown in the cool northern parts of the United States were investigated. Individual phenolic compounds, including phenolic acids, flavan-3-ols, flavones, and anthocyanins, were further quantitatively investigated by HPLC. Cellular antioxidant activities (CAA) and peroxyl radical scavenging capacity (PRSC) were evaluated by fluorescence microplate reader. Cultivar Morton exhibited the highest individual flavan-3-ols (catechin and epicatechin) and total flavonoids, as well as the highest antioxidant properties (PRSC and CAA) among all lentils tested. Five phenolic acids of the benzoic types and their derivates (gallic, protocatechuic, 2,3,4-trihydroxybenzoic, p-hydroxybenzoic acid, and protocatechualdehyde) and four phenolic acids of the cinnamic type (chlorogenic, p-coumaric, m-coumaric, and sinapic acid) were detected in all lentil cultivars. Two flavan-3-ols [(+)-catechin and (-)-epicatechin] and one flavone (luteolin) were detected in all lentil cultivars. Among all phenolic compounds detected, sinapic acid was the predominant phenolic acid, and (+)-catechin and (-)-epicatechin were the predominant flavonoids. These results showed that different phenotype lentils possessed considerable variations in their individual phenolic compounds, as well as chemical and cellular antioxidant activities. Caffeic acid, catechin, epicatechin, and total flavonoids significantly (p < 0.05) correlated with peroxyl radical scavenging assay. Cellular antioxidant assay significantly correlated with chemical antioxidant assay ORAC. The results from this study could be very interesting for breeding programs to improve lentils for use as functional foods.

  19. Chemical Composition, Antibacterial and Phytotoxic Activities of Peganum harmala Seed Essential Oils from Five Different Localities in Northern Africa.

    PubMed

    Apostolico, Ida; Aliberti, Luigi; Caputo, Lucia; De Feo, Vincenzo; Fratianni, Florinda; Nazzaro, Filomena; Souza, Lucèia Fàtima; Khadhr, Maroua

    2016-09-15

    Peganum harmala L., also known as Syrian rue or Pègano, is a herbaceous plant belonging to the Zygohpyllaceae family, and is widely used in traditional medicine. The chemical composition of essential oils of P. harmala seeds from five different regions of Northern Africa (Algeria, Egypt, Libya, Morocco and Tunisia) was studied by GC and GC-MS analyses. A total of 105 compounds were identified, the main components being oxygenated monoterpenes and oxygenated sesquiterpenes. Eugenol is the main component in all oils. The antimicrobial activity of the essential oils was assayed against some bacterial strains: Staphylococcus aureus (DSM 25693), Bacillus cereus (DSM 4313), Bacillus cereus (DSM4384), Escherichia coli (DMS 857) and Pseudomonas aeruginosa (ATCC 50071). All the oils showed different inhibitory activity. In the twentieth century this is an important result; we need possible new botanical drugs because the problem of resistance to antimicrobial drugs has become apparent. Moreover, the essential oils were evaluated for their possible in vitro phytotoxic activity against germination and initial radicle growth of Raphanus sativus L., Lepidium sativum L., and Ruta graveolens L. The results showed that both germination and radical elongation were sensitive to the oils.

  20. Repeated large-magnitude earthquakes in a tectonically active, low-strain continental interior: The northern Tien Shan, Kyrgyzstan

    NASA Astrophysics Data System (ADS)

    Landgraf, A.; Dzhumabaeva, A.; Abdrakhmatov, K. E.; Strecker, M. R.; Macaulay, E. A.; Arrowsmith, Jr.; Sudhaus, H.; Preusser, F.; Rugel, G.; Merchel, S.

    2016-05-01

    The northern Tien Shan of Kyrgyzstan and Kazakhstan has been affected by a series of major earthquakes in the late 19th and early 20th centuries. To assess the significance of such a pulse of strain release in a continental interior, it is important to analyze and quantify strain release over multiple time scales. We have undertaken paleoseismological investigations at two geomorphically distinct sites (Panfilovkoe and Rot Front) near the Kyrgyz capital Bishkek. Although located near the historic epicenters, both sites were not affected by these earthquakes. Trenching was accompanied by dating stratigraphy and offset surfaces using luminescence, radiocarbon, and 10Be terrestrial cosmogenic nuclide methods. At Rot Front, trenching of a small scarp did not reveal evidence for surface rupture during the last 5000 years. The scarp rather resembles an extensive debris-flow lobe. At Panfilovkoe, we estimate a Late Pleistocene minimum slip rate of 0.2 ± 0.1 mm/a, averaged over at least two, probably three earthquake cycles. Dip-slip reverse motion along segmented, moderately steep faults resulted in hanging wall collapse scarps during different events. The most recent earthquake occurred around 3.6 ± 1.3 kyr ago (1σ), with dip-slip offsets between 1.2 and 1.4 m. We calculate a probabilistic paleomagnitude to be between 6.7 and 7.2, which is in agreement with regional data from the Kyrgyz range. The morphotectonic signals in the northern Tien Shan are a prime example of deformation in a tectonically active intracontinental mountain belt and as such can help understand the longer-term coevolution of topography and seismogenic processes in similar structural settings worldwide.

  1. Implications of climatic seasonality on activity patterns and resource use by sympatric peccaries in northern Pantanal.

    PubMed

    Hofmann, Gabriel Selbach; Coelho, Igor Pfeifer; Bastazini, Vinicius Augusto Galvão; Cordeiro, José Luís Passos; de Oliveira, Luiz Flamarion Barbosa

    2016-03-01

    We evaluated the effects of climate seasonality from a thermal and water availability perspective on the activity patterns and resource use of Pecari tajacu and Tayassu pecari during wet and dry seasons in the northeastern Brazilian Pantanal. We used camera traps and temperature sensors to record species activity patterns in relation to temperature, established five habitat categories based on flooding intensity and local vegetation characteristics, assessed the activity patterns of each species in dry and wet periods and in artificial water bodies using circular statistical metrics, and calculated niche amplitude and overlap on three axes (temperature, time, and habitat) in both periods. Peccaries shared a strong resemblance in resource use and in their responses to seasonal variations in the tested gradients. The activity patterns of both species exhibited a significant correlation with air temperature on all the evaluated measures, and both species strongly reduced their activity when the air temperature exceeded 35 °C. High temperatures associated with low water availability were most likely responsible for the changes in species activity patterns, which resulted in an increased temporal overlap in habitat use throughout the dry season. However, the peccaries avoided intensively flooded habitats; therefore, the habitat gradient overlap was greater during the wet period. Our results show that an increase in niche overlap on the environmental gradient as a result of climatic seasonality may be partially compensated by a reduction in other niche dimensions. In this case, temporal partitioning appears to be an important, viable mechanism to reduce competition by potentially competing species.

  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. Retention and sustainability of community-based health volunteers' activities: A qualitative study in rural Northern Ghana

    PubMed Central

    Chatio, Samuel; Akweongo, Patricia

    2017-01-01

    Background The shortage of formal health workers has led to the utilization of Community-Based Health Volunteers (CBHV) to provide health care services to people especially in rural and neglected communities. Community-based health volunteers have been effective partners in health care delivery at the community level for many years. The challenge is how to retain these volunteers and also sustain their activities. This study explored factors affecting retention and sustainability of community-based health volunteers’ activities in a rural setting in Northern Ghana. Methods This was a qualitative study comprising thirty-two in-depth interviews (IDIs) with health volunteers and health workers in-charge of health volunteers’ activities. Purposive sampling technique was used to select study participants for the interviews. The interviews were transcribed and coded into themes using Nvivo 10 software. The thematic analysis framework was used to analyze the data. Results Study participants reported that the desire to help community members, prestige and recognition as doctors in community mainly motivated them to work as health volunteers. Lack of incentives and logistical supplies such as raincoats, torch lights, wellington boots and transportation in the form of bicycles to facilitate the movement of health volunteers affected the work. They suggested that lack of these things discouraged them from working as health volunteers. Most of the dropout volunteers said lack of support and respect from community members made them to stop working as health volunteers. They recommended that community support, incentives and logistical supplies such as raincoats, torch light, wellington boots, bicycles, awards to hard working volunteers are mechanisms that can help retain community-based health volunteers and also sustain their activities. Conclusion Providing means of transport and non-monetary incentives would help to retain community-based health volunteers and also

  4. Paleozoic evolution of active margin basins in the southern Central Andes (northwestern Argentina and northern Chile)

    NASA Astrophysics Data System (ADS)

    Bahlburg, H.; Breitkreuz, C.

    The geodynamic evolution of the Paleozoic continental margin of Gondwana in the region of the southern Central Andes is characterized by the westward progression of orogenic basin formation through time. The Ordovician basin in the northwest Argentinian Cordillera Oriental and Puna originated as an Early Ordovician back-arc basin. The contemporaneous magmatic arc of an east-dipping subduction zone was presumably located in northern Chile. In the back-arc basin, a ca. 3500 meter, fining-up volcaniclastic apron connected to the arc formed during the Arenigian. Increased subsidence in the late Arenigian allowed for the accomodation of large volumes of volcaniclastic turbidites during the Middle Ordovician. Subsidence and sedimentation were caused by the onset of collision between the para-autochthonous Arequipa Massif Terrane (AMT) and the South American margin at the Arenigian-Llanvirnian transition. This led to eastward thrusting of the arc complex over its back-arc basin and, consequently, to its transformation into a marine foreland basin. As a result of thrusting in the west, a flexural bulge formed in the east, leading to uplift and emergence of the Cordillera Oriental shelf during the Guandacol Event at the Arenigian-Llanvirnian transition. The basin fill was folded during the terminal collision of the AMT during the Oclóyic Orogeny (Ashgillian). The folded strata were intruded post-tectonically by the presumably Silurian granitoids of the "Faja Eruptiva de la Puna Oriental." The orogeny led to the formation of the positive area of the Arco Puneño. West of the Arco Puneño, a further marine basin developed during the Early Devonian, the eastern shelf of which occupied the area of the Cordillera Occidental, Depresión Preandina, and Precordillera. The corresponding deep marine turbidite basin was located in the region of the Cordillera de la Costa. Deposition continued until the basin fill was folded in the early Late Carboniferous Toco Orogeny. The basin

  5. 10 years of cloud droplet activation data from Pallas, Northern Finland - preliminary results

    NASA Astrophysics Data System (ADS)

    Kivekäs, Niku; Asmi, Eija; Brus, David; Komppula, Mika; Lihavainen, Heikki

    2016-04-01

    Activation of atmospheric aerosol particles into cloud droplets has been studied in situ at Pallas measurement station in Finnish Lapland from year 2005 to present day. The site is located on a hill top, about 300 m above the surrounding lowlands, and it is inside a cloud for 15 % of time. Here in-cloud periods are defined as periods when visibility was below 1000 m. There are two parallel Differential Mobility Particle Sizers (DMPS) at the site, measuring the number concentration and dry size distribution of atmospheric aerosol particles. One DMPS is connected to a PM2.5 inlet, the other to a total air inlet with no cut-off diameter. After each inlet the particles are dried to evaporate any water in them. This way it is possible to measure simultaneously the dry number-size-distribution of all particles, and that of particles with wet diameter smaller than 2.5 m. As the latter does not include cloud droplets, the difference between the two measurements represents the number concentration and size distribution of those particles that have activated into cloud droplets. The number concentration of particles at Pallas has a clear seasonal cycle, being highest during summer and lowest during winter. The monthly mean number concentration of particles with diameter larger than 100 nm varied from 38 cm-3 in November to 270 cm-3 in July. During in-cloud periods the monthly mean number concentration of activated particles of this same size class showed a similar pattern, varying from 23 cm-3 (November) to 110 cm-3 in April. The monthly mean D50 activation diameter (diameter at which 50 % of particles activate) varied from 85 nm (February) to 189 nm (July), showing an average 0.1 nm increase for each added particle with diameter > 100 nm. The activated fraction of particles in all sizes decreased sharply when visibility exceeded 1000 m. The highest activated fractions of particles were not observed during the periods of the thickest clouds, but during clouds with in

  6. 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.

  7. 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

  8. Physical Education Teacher Attitudes towards the Effectiveness of Sport Activities in Northern-East Badia

    ERIC Educational Resources Information Center

    Al-Oun, Ismael Sood

    2015-01-01

    The purpose of this study is to investigate physical education teacher attitudes towards the effectiveness of sport activities in north-east Badia. The study population consisted of all teachers of physical education who are studying curriculum of physical education in schools affiliated to the Directorate of Education of the brigade desert…

  9. The polyphenol content and antioxidant activities of the main edible vegetables in northern Vietnam.

    PubMed

    Thu, Nghiem Nguyet; Sakurai, Chika; Uto, Harumi; Van Chuyen, Nguyen; Lien, Do Thi Kim; Yamamoto, Shigeru; Ohmori, Reiko; Kondo, Kazuo

    2004-06-01

    Oxidized low-density lipoprotein (LDL) is considered a risk factor in atherosclerosis, and polyphenols are the potential agents to inhibit the oxidation of LDL. We determined the polyphenol contents and the antioxidant activities of commonly consumed vegetables in Vietnam and assessed the quantity of the polyphenol intake from vegetables in the current Vietnamese diet. The polyphenol contents in 30 kinds of vegetables was determined by the Folin-Ciocalteu method. The antioxidant activities of vegetables were evaluated by measuring the oxidation of LDL and the reduction of the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. In this study, some herbs and edible wild vegetables possessed high contents of polyphenols and antioxidant activities. Among green vegetables, sweet potato leaves showed both a high polyphenol content and antioxidant activity. The mean polyphenol daily intake of the Vietnamese was a 595 mg catechin equivalent. Water spinach, a kind of green vegetable, contributed the highest amount (45%) of the total polyphenol intake, followed by other green vegetables. Neither herbs nor edible wild vegetables contributed significantly to the total polyphenol intake due to their low consumption. Green vegetables are therefore considered very important sources of polyphenol intake for the Vietnamese.

  10. School Engagement among Aboriginal Students in Northern Canada: Perspectives From Activity Settings Theory

    ERIC Educational Resources Information Center

    Davison, Colleen M.; Hawe, Penelope

    2012-01-01

    Background: Educational disengagement is a public health concern among Aboriginal populations in many countries. It has been investigated previously in a variety of ways, with the conventional focus being on the children themselves. Activity settings are events and places, theorized in terms of their symbols, roles, time frame, funds, people, and…

  11. Bat mortality and activity at a Northern Iowa wind resource area

    USGS Publications Warehouse

    Jain, A.A.; Koford, Rolf R.; Hancock, A.W.; Zenner, G.G.

    2011-01-01

    We examined bat collision mortality, activity and species composition at an 89-turbine wind resource area in farmland of north-central Iowa from mid-Apr. to mid-Dec., 2003 and mid-Mar. to mid-Dec., 2004. We found 30 bats beneath turbines on cleared ground and gravel access areas in 2003 and 45 bats in 2004. After adjusting for search probability, search efficiency and scavenging rate, we estimated total bat mortality at 396 ?? 72 (95 ci) in 2003 and 636 ?? 112 (95 ci) in 2004. Although carcasses were mostly migratory tree bats, we found a considerable proportion of little brown bats (Myotis lucifugus). We recorded 1465 bat echolocation call files at turbine sites ( 34.88 call files/detector-night) and 1536 bat call files at adjacent non-turbine sites ( 36.57 call files/detector-night). Bat activity did not differ significantly between turbine and non-turbine sites. A large proportion of recorded call files were made by Myotis sp. but this may be because we detected activity at ground level only. There was no relationship between types of turbine lights and either collision mortality or echolocation activity. The highest levels of bat echolocation activity and collision mortality were recorded during Jul. and Aug. during the autumn dispersal and migration period. The fatality rates for bats in general and little brown bats in particular were higher at the Top of Iowa Wind Resource Area than at other, comparable studies in the region. Future efforts to study behavior of bats in flight around turbines as well as cumulative impact studies should not ignore non-tree dwelling bats, generally regarded as minimally affected. ?? 2011, American Midland Naturalist.

  12. Disseminated sulphides in basalts from the northern Central Indian Ridge: implications on late-stage hydrothermal activity

    NASA Astrophysics Data System (ADS)

    Banerjee, Ranadip; Ray, Dwijesh

    2015-04-01

    This study examined the mineralogy and mineral chemistry of disseminated sulphides (mainly chalcopyrite-pyrite) in partly altered basalts from the northern Central Indian Ridge, Indian Ocean in order to understand the role of hydrothermal alterations and infer possible sulphide formation history. Pyrite and chalcopyrite are dominant sulphide minerals and generally associated with the oxide phases including magnetite and often ilmenite. Close association of sulphide and oxide minerals suggests that they are paragenetically related. Sulphides also occur as late impregnated veins cutting through the basaltic hosts. The chemical compositions of pyrite (avg. Fe 46.3 wt%, S 53.7 wt%) and chalcopyrite (avg. Cu 34.4 wt%, Fe 30.7 wt%, S 34.7 wt%) are almost uniform, while the secondary ilmenite often shows MnO enrichment (up to 3.0-3.4 wt%). The associated altered minerals typically resemble the greenschist facies mineral assemblages—e.g. chlorite±epidote. Evidence of albitisation and silicification suggests low-temperature hydrothermal alteration processes. This is supported by the bulk Au content (up to 60 ppb) of host-altered basalts with pyrite mineralisation. Au is usually associated with late-stage pyrites and thus related with low-temperature hydrothermal activity. Close to the dredge location, tectonic activity around the Vityaz megamullion might have promoted hydrothermal circulation and subsequent alteration of the mineral constituents in basalts, eventually inducing the formation of late-stage disseminated sulphide minerals in these rocks.

  13. Structural characteristics of an active fold-and-thrust system in the southeastern Atacama Basin, northern Chile

    NASA Astrophysics Data System (ADS)

    Lin, Yen-Sheng; Chuang, Yi-Rung; Shyu, J. Bruce H.; González, Gabriel; Shen, Chuan-Chou; Lo, Ching-Hua; Liou, Ya-Hsuan

    2016-08-01

    The western South American margin is one of the most active plate boundaries in the world. Using various remote sensing data sets, we mapped the neotectonic characteristics of an area at the southeastern corner of the Atacama Basin, northern Chile, in the Andean forearc. There, one major N-S trending ridge is clearly visible both in the satellite images and in the field. This ridge reaches 250 m above the basin floor in its middle part and is asymmetrical, with a steep eastern slope and a much gentler western slope. The geometry of the ridge indicates that it formed as an asymmetrical anticline. This anticline is likely formed as a shear fault-bend fold, with a major décollement at a depth of about 2.5 km in the Naranja Formation. We suggest that this décollement is a major structure of the Atacama Basin area. From the ages of the ignimbrites and lake deposits that were deformed by this anticline, we obtained a long-term shortening rate of the major underlying structure at about 0.2 mm/yr. This thin-skinned fold-and-thrust system appears to be active since at least about 3 Ma, and could be as long as since middle Miocene. Therefore, crustal structures may play important roles in the Neogene development of the western Andean margin.

  14. Quaternary Uplift of Coral Terraces from Active Folding and Thrusting Along the Northern Coast of Timor-Leste

    NASA Astrophysics Data System (ADS)

    Cox, N. L.; Harris, R.; Merritts, D.

    2006-12-01

    Emergent coral terraces along the northern coast of Timor-Leste, reveal differential vertical strain along the strike of the active Banda arc-continent collision complex. The number of major coral terraces in surveyed profiles increases from 2 to 25 along a coastal distance of 150 km from central to eastern Timor-Leste. Comparison of 8 separate terrace profiles with sea level curves predicts vertical displacement rates that increase eastward from <0.3 to 1.0-1.5 mm/year. This pattern is corroborated by U-series age analyses that document non-linear increases in vertical strain eastward with the uplift rates ranging from <.04 to 2.0 mm/year. U-series ages also indicates the occurrence of both erosional (regressional) and depositional terraces. Two profiles, dominated by erosional terraces, have associated local lower depositional terraces. The erosional terraces are more abundant, but yield apparent lower uplift rates in comparison with the depositional terraces. This occurrence questions the validity of uplift rates obtained by age analysis for the errosional terraces and/or aids in the conclusion that differential vertical strain not only exists with distance between profiles but with time for each profile. We associate active uplift with northward movement along retro- wedge thrust faults that are well documented in seismic reflection profiles adjacent to the north coast of Timor- Leste.

  15. Effect of geomagnetic activity on the northern annular mode: QBO dependence and the Holton-Tan relationship

    NASA Astrophysics Data System (ADS)

    Maliniemi, V.; Asikainen, T.; Mursula, K.

    2016-09-01

    Mutually conflicting results have been presented in earlier studies on the long-term relation of geomagnetic activity (GA) and the winter northern annular mode (NAM) and its modulation by quasi-biennial oscillation (QBO). Some studies have found a stronger positive relation in the easterly phase of the QBO, while in other studies a stronger positive relation was found in the westerly phase of the QBO. Using QBO reconstruction from the beginning of the twentieth century we find that the QBO modulation of the GA-NAM relation is temporally variable, which explains the earlier, seemingly differing results. Positive relation is found to be valid in the easterly QBO phase at 30 hPa for the whole twentieth century. We also find that the QBO at 30 hPa better represents the Holton-Tan relation for the surface circulation and that the Holton-Tan relation is only observed during early/mid winter, while an anti-Holton-Tan relation is found in the late winter for strong geomagnetic activity. These results emphasize the systematic response of NAM to particle precipitation during the entire twentieth century and underline the importance of considering the preconditioning of the atmosphere when studying the solar-related effects upon climate.

  16. 5'AMP-activated protein kinase activity is increased in adipose tissue of northern elephant seal pups during prolonged fasting-induced insulin resistance.

    PubMed

    Viscarra, Jose A; Champagne, Cory D; Crocker, Daniel E; Ortiz, Rudy M

    2011-06-01

    Northern elephant seals endure a 2- to 3-month fast characterized by sustained hyperglycemia, hypoinsulinemia, and increased plasma cortisol and free fatty acids, conditions often seen in insulin-resistant humans. We had previously shown that adipose Glut4 expression and 5'AMP-activated protein kinase (AMPK) activity increase and plasma glucose decreases in fasting seals suggesting that AMPK activity contributes to glucose regulation during insulin-resistant conditions. To address the hypothesis that AMPK activity increases during fasting-induced insulin resistance, we performed glucose tolerance tests (GTT) on early (n=5) and late (n=8)-fasted seal pups and compared adipose tissue expression of insulin signaling proteins, peroxisome proliferator-activated receptor γ (PPARγ), and AMPK, in addition to plasma adiponectin, leptin, cortisol, insulin, and non-esterified fatty acid (NEFA) levels. Fasting was associated with decreased glucose clearance, plasma insulin and adiponectin, and intracellular insulin signaling, as well as increased plasma cortisol and NEFAs, supporting the suggestion that seals develop insulin resistance late in the fast. The expression of Glut4 and VAMP2 increased (52 and 63% respectively) with fasting but did not change significantly during the GTT. PPARγ and phosphorylated AMPK did not change in the early fasted seals, but increased significantly (73 and 50% respectively) in the late-fasted seals during the GTT. Increased AMPK activity along with the reduction in the activity of insulin-signaling proteins supports our hypothesis that AMPK activity is increased following the onset of insulin resistance. The association between increased AMPK activity and Glut4 expression suggests that AMPK plays a greater role in regulating glucose metabolism in mammals adapted to prolonged fasting than in non-fasting mammals.

  17. 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.

  18. The elevational pattern of microbial community and enzyme activity along the northern slop of Changbai Mountain

    NASA Astrophysics Data System (ADS)

    Xu, Zhiwei; Yu, Guirui; Zhang, Xinyu; Ge, Jianpin; He, Nianpeng; Wang, Qiufeng; Wang, Dan

    2014-05-01

    we present a comprehensive analysis of soil microbial community structure, enzyme activities and their role in soil organic matter mineralization along six elevations representing five typical vegetation types from forest to alpine tundra in Changbai Mountain, China. The results showed that the microbial PLFAs presented hump-shaped patterns along the elevation with the total microbial PLFAs highest in Ermans birch forest soil. The fungi to bacteria and gram positive to negative bacteria ratios increased along the elevation with the lowest values in Broad leaved forest and Dark-coniferous spruce-fir forest soil, respectively. The soil microbial community structures showed a biogeography distribution pattern in vertical direction with microbial community structures in Broad leaved forest and Mixed coniferous broad leaved forest different from other four sites. The soil enzyme activities in Broad leaved forest and Mixed coniferous broad leaved forest were significantly higher than in other four elevations. Principal component analysis (PCA) revealed substantial differences in soil microbial community composition among study sites, appeared to be driven primarily by MAT, MAP, soil temperature and content of silt & clay on the first principal component (PC1) which accounted for 87.1 % of the total sample variance. However, soil nutrients mainly responsible for the variation of soil enzyme activities. The soil organic matter mineralization rates tended to be highest in Ermans birch forest site and lowest in Dark-coniferous spruce-fir forest site and showed positive relationship with total microbial, bacterial and actinomycetes PLFAs. These findings could be used to facilitate interpretation of soil microbial community and ecological function in latitude forests ecosystem especially in volcanic forest ecosystem.

  19. Active tectonics of the Devils Mountain Fault and related structures, northern Puget Lowland and eastern Strait of Juan de Fuca region, Pacific Northwest

    USGS Publications Warehouse

    Johnson, Samuel Y.; Dadisman, Shawn V.; Mosher, David C.; Blakely, Richard J.; Childs, Jonathan R.

    2001-01-01

    Information from marine high-resolution and conventional seismic-reflection surveys, aeromagnetic mapping, coastal exposures of Pleistocene strata, and lithologic logs of water wells is used to assess the active tectonics of the northern Puget Lowland and eastern Strait of Juan de Fuca region of the Pacific Northwest. These data indicate that the Devils Mountain Fault and the newly recognized Strawberry Point and Utsalady Point faults are active structures and represent potential earthquake sources.

  20. Middle to Late Pleistocene activity of the northern Matese fault system (southern Apennines, Italy)

    NASA Astrophysics Data System (ADS)

    Galli, Paolo; Giaccio, Biagio; Messina, Paolo; Peronace, Edoardo; Amato, Vincenzo; Naso, Giuseppe; Nomade, Sebastian; Pereira, Alison; Piscitelli, Sabatino; Bellanova, Jessica; Billi, Andrea; Blamart, Dominique; Galderisi, Antonio; Giocoli, Alessandro; Stabile, Tony; Thil, Francoise

    2017-03-01

    An integrated investigation including geological, geomorphological, geophysical and structural survey, tephra analyses, 14C and 40Ar/39Ar dating, as well as paleoseismic trenching along the N-Matese fault system is presented. The study allowed the characterization of the tectonic mobility of this structure as well as the associated Bojano basin sedimentary-tectonic evolution since the early Middle Pleistocene, providing also new clues concerning the fault historical activity and the associated Mw > 6.5 earthquakes. We have found lines of evidence for > 1 mm/yr slip rate along the presently buried Bojano fault during the mid Middle Pleistocene, and similar rates for the main fault segments paralleling the Matese flanks. The buried Bojano fault significantly slowed down during the last 300 kyr, ceasing its activity before the Holocene. In turn, the segments outcropping along the Matese flanks reactivated at the onset of Late Pleistocene, after a long period of quiescence (480-110 ka), with robust slip rates that would seem even accelerating in post LGM times. Paleoseismic data suggest the occurrence of four Mw > 6.6 earthquakes in the past 3 ka, three of which match the little known 280 BC event, and the devastating 1456 and 1805 earthquakes.

  1. Influence of season on daytime behavioral activities of donkeys in the Northern Guinea Savanna zone of Nigeria

    PubMed Central

    ZAKARI, Friday Ocheja; AYO, Joseph Olusegun; REKWOT, Peter Ibrahim; KAWU, Mohammed Umar

    2016-01-01

    ABSTRACT The present experiment was performed with the aim of investigating the effect of season on behavioral activities of donkeys during the rainy and harmattan seasons in the Northern Guinea zone of Nigeria. Sixteen apparently healthy donkeys were used as subjects and divided into four groups based on age. During each season, behavioral activities of each donkey were evaluated for three weeks using the focal animal sampling technique. The dry-bulb temperature (DBT), relative humidity (RH), and temperature-humidity index (THI) were obtained three times each day during the experimental period using standard procedures. In the rainy season, the mean DBT (31.65 ± 0.49°C), RH (73.63 ± 1.09%), and THI (84.39 ± 0.71) were significantly (P<0.0001) higher than the corresponding values of 24.00 ± 0.44°C, 36.80 ± 0.92%, and 64.80 ± 0.62 in the harmattan season. During the rainy season, the donkeys spent 60.00 ± 0.77%, 25.40 ± 0.69%, and 2.94 ± 0.21% on grazing, resting, and grooming, respectively. During the harmattan season, the donkeys spent the most time on grazing (76.76 ± 0.43%), less time on resting (11.97 ± 0.38%), and the least time on grooming (0.89 ± 0.05%). In conclusion, season and seasonal variations affect the daytime behavioral activities of donkeys in the zone, and this should be considered in husbandry practices for donkeys. PMID:26858575

  2. Geochemical characterization and biomonitoring of reclaimed soils in the Po River Delta (Northern Italy): implications for the agricultural activities.

    PubMed

    Di Giuseppe, Dario; Bianchini, Gianluca; Vittori Antisari, Livia; Martucci, Annalisa; Natali, Claudio; Beccaluva, Luigi

    2014-05-01

    This geochemical study is focused on the easternmost part of the Po River alluvial plain in Northern Italy, which is interested by widespread agricultural activities, investigating a reclaimed sector of the Province of Ferrara, known as "Valle del Mezzano" (Mezzano Low Land, hereafter reported as MLL) characterized by peat-rich soils. The chemical-mineralogical characterization of these reclaimed soils is important to compare the local geochemical backgrounds with those recorded in other sectors of the River Po plain and to monitor if the observed concentration exceeds critical thresholds. The reported analyses include (a) measurement of the soil salinity, (b) nutrient evaluation, (c) major and trace element concentrations carried out on bulk soils, (d) tests of metal extraction with both aqua regia and EDTA to highlight the distinct elemental mobility and (e) phyto-toxicological measurement of heavy metal concentrations in plants (Lactuca sativa acephala) grown on the studied soils. The results indicate (1) high soil salinity, often with drastic increase of sodium and chloride along the soil profiles, (2) high nitrogen content (in part related to anthropogenic activities) on superficial horizons and nitrate decrease along the soil profiles and (3) comparative enrichments in heavy metals with respect to other soils of the province, which indicate that peat deposits are effective in trapping metals from anthropogenic sources. This, in turn, implies potential geochemical risks for the agricultural activities. In this regard, specific concerns are related to the high nickel and arsenic content of MLL soils due to the mobility of these elements and their attitude to be taken up by plants.

  3. Influence of season on daytime behavioral activities of donkeys in the Northern Guinea Savanna zone of Nigeria.

    PubMed

    Zakari, Friday Ocheja; Ayo, Joseph Olusegun; Rekwot, Peter Ibrahim; Kawu, Mohammed Umar

    2015-01-01

    The present experiment was performed with the aim of investigating the effect of season on behavioral activities of donkeys during the rainy and harmattan seasons in the Northern Guinea zone of Nigeria. Sixteen apparently healthy donkeys were used as subjects and divided into four groups based on age. During each season, behavioral activities of each donkey were evaluated for three weeks using the focal animal sampling technique. The dry-bulb temperature (DBT), relative humidity (RH), and temperature-humidity index (THI) were obtained three times each day during the experimental period using standard procedures. In the rainy season, the mean DBT (31.65 ± 0.49°C), RH (73.63 ± 1.09%), and THI (84.39 ± 0.71) were significantly (P<0.0001) higher than the corresponding values of 24.00 ± 0.44°C, 36.80 ± 0.92%, and 64.80 ± 0.62 in the harmattan season. During the rainy season, the donkeys spent 60.00 ± 0.77%, 25.40 ± 0.69%, and 2.94 ± 0.21% on grazing, resting, and grooming, respectively. During the harmattan season, the donkeys spent the most time on grazing (76.76 ± 0.43%), less time on resting (11.97 ± 0.38%), and the least time on grooming (0.89 ± 0.05%). In conclusion, season and seasonal variations affect the daytime behavioral activities of donkeys in the zone, and this should be considered in husbandry practices for donkeys.

  4. 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…

  5. 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.

  6. [Regularity of psychophysiological adaptation to military-professional activity among staff of Northern Fleet].

    PubMed

    Sementsov, V K; Khankevich, Iu R; Mosiagin, I G; Maruniak, S V; Chesnokova, V N

    2010-03-01

    There were examined regularities of psycho-physiological adaptation of different categories of military-navy specialists, were detected periods of maximum decrease of compensation abilities among sailor-men and staff reserve of Naval Fleet from students. Were educed psycho-physiological strategies of adaptation of military-navy specialists in dependence of reference level of adaptation capabilities. Was detected the influence on the process of psycho-physiological adaptation to service activity of young specialists of a complex of psychological and social factors. Was educed the influence of ulterior motive on success of psycho-physiological adaptation to new conditions of environment. Was elaborated a complex of measures on optimization of accompaniment of adaptation of military-navy specialists and was given financial evaluation of it's implementation.

  7. Recent activity of Anatahan volcano, Northern Marina Islands, and its magma plumbing system

    NASA Astrophysics Data System (ADS)

    Nakada, S.; Morita, Y.; Matsushima, T.; Tabei, T.; Watanabe, A.; Maeno, F.; Camacho, J. T.

    2009-12-01

    The volcanic activity of Anatahan that began in 2003 has declined such as faint emission of volcanic gas from the crater and scarcity of volcanic tremor in 2009. Our team carried out geological, geodetic and seismological observation repeatedly till mid-2009 from the beginning of the eruption. The early phase of the eruption (2003-2004) can be characterized by magmatic and phreatomagmatic explosions, contrasting to mainly phreatic nature in the later phase (2005-2008). The active crater (Eastern Crater) was widened and deepened (much below the sea level) as the eruption progressed. Dominant products of phreatic explosions comprise of thick accumulation of thin layers of fine ash. A rough estimate of the total volume during these 5 years is as much as 1 km3, close to the volume of materials lost by enlargement of the active crater. Seismic observation was carried out during mid-2008 and mid-2009 by settling 5 temporary stations covering the whole of the island, each of which includes a 3 components short-period seismometer with corner frequency of 1Hz and a low-power consumption digital data recorder with 24-bits AD resolutions. GPS campaign observation was repeated in the same station during this period. VT and LP event were observed, though very low in occurrence in this period. Hypocenters of VT and LP events show all events occurred at the depth of less than 8km around the eastern crater. Among them, LP events occurred in the shallower (less than 3km) region. The error in the depth may be not more than a few kilometers, but that in the epicenter should be smaller than 1km because the most events are located inside of the seismic network. Moreover, the tremors observed in the 2008 summer continued for about 3 weeks. The amplitude increased gradually, kept at the maximum, and stopped abruptly. During the maximum amplitude period, ash emission was observed by VAAC. Estimated reduced displacement at the maximum is about 1 cm2, typical of a hydro-magmatic eruption

  8. Response of extratropical cyclone activity to the Kuroshio large meander in northern winter

    NASA Astrophysics Data System (ADS)

    Hayasaki, Masamitsu; Kawamura, Ryuichi; Mori, Masato; Watanabe, Masahiro

    2013-06-01

    We examined possible responses of cyclone activities to the bimodal path states of the Kuroshio Current [i.e., large meander (LM) and non-LM (NLM)] by using the long-term reanalysis data and the 20th century hindcast experiment of a high-resolution atmosphere-ocean coupled model. Compared with a seasonal mean cyclone track frequency for the LM and NLM periods, a primary cyclone track shifts southward in association with the meander of Kuroshio Current. Composite analyses of the hindcast experiment showed remarkable atmospheric responses accompanying the Kuroshio LM. The Kuroshio LM causes a decrease in latent heat flux in the south of Japan and a southward shift of the near-surface baroclinic zone. Distinctive decreases in thermodynamic fluxes inhibit the rapid development of cyclones in the meander region, eventually inducing positive sea level pressure anomalies downstream from that region.

  9. The dynamic relation between activities in the Northern and Southern solar hemispheres

    NASA Astrophysics Data System (ADS)

    Volobuev, D. M.; Makarenko, N. G.

    2016-12-01

    The north-south (N/S) asymmetry of solar activity is the most pronounced phenomenon during 11-year cycle minimums. The goal of this work is to try to interpret the asymmetry as a result of the generalized synchronization of two dynamic systems. It is assumed that these systems are localized in two solar hemispheres. The evolution of these systems is considered in the topological embeddings of a sunspot area time series obtained with the use of the Takens algorithm. We determine the coupling measure and estimate it on the time series of daily sunspot areas. The measurement made it possible to interpret the asymmetry as an exchangeable dynamic equation, in which the roles of the driver-slave components change in time for two hemispheres.

  10. 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.

  11. Gauging state-level and user group views of oyster reef restoration activities in the northern Gulf of Mexico

    USGS Publications Warehouse

    LaPeyre, Megan K.; Nix, Ashby; Laborde, Luke; Piazza, Bryan P.

    2012-01-01

    Successful oyster reef restoration, like many conservation challenges, requires not only biological understanding of the resource, but also stakeholder cooperation and political support. To measure perceptions of oyster reef restoration activities and priorities for future restoration along the northern Gulf of Mexico coast, a survey of 1500 individuals representing 4 user groups (oyster harvesters, shrimpers, environmental organization members, professionals), across 5 states (Texas, Louisiana, Mississippi, Alabama, Florida) was conducted in 2011. All respondents highly supported reef restoration efforts, but there was a dichotomy in preferred restoration goals with commercial fishermen more likely to support oyster reef restoration for stock enhancement, while professionals and environmental organization members were more likely to support oyster reef restoration to enhance ecosystem services. All user groups identified enforcement, funding, and appropriate site selection as basic requirements for successful reef restoration. For management of restored oyster reefs, oyster harvesters and shrimpers were less likely to support options that restricted the use of reefs, including gear restrictions and permanent closures, but did support rotating annual reef closures, while other stakeholders were willing to consider all options, including annual reef closures and sanctuary reefs. Overall, there were clear differences in management and communication preferences across user groups, but few differences across states. Understanding these key differences in stakeholder support for, and willingness to accept specific management actions is critical in moving management and restoration forward while minimizing conflict.

  12. High-resolution shallow reflection seismic image and surface evidence of the Upper Tiber Basin active faults (Northern Apennines, Italy)

    USGS Publications Warehouse

    Donne, D.D.; Plccardi, L.; Odum, J.K.; Stephenson, W.J.; Williams, R.A.

    2007-01-01

    Shallow seismic reflection prospecting has been carried out in order to investigate the faults that bound to the southwest and northeast the Quaternary Upper Tiber Basin (Northern Apennines, Italy). On the northeastern margin of the basin a ??? 1 km long reflection seismic profile images a fault segment and the associated up to 100 meters thick sediment wedge. Across the southwestern margin a 0.5 km-long seismic profile images a 50-55??-dipping extensional fault, that projects to the scarp at the base of the range-front, and against which a 100 m thick syn-tectonic sediment wedge has formed. The integration of surface and sub-surface data allows to estimate at least 190 meters of vertical displacement along the fault and a slip rate around 0.25 m/kyr. Southwestern fault might also be interpreted as the main splay structure of regional Alto Tiberina extensional fault. At last, the 1917 Monterchi earthquake (Imax=X, Boschi et alii, 2000) is correlable with an activation of the southwestern fault, and thus suggesting the seismogenic character of this latter.

  13. Distribution of zooplankton biomass and potential metabolic activities across the northern Benguela upwelling system

    NASA Astrophysics Data System (ADS)

    Fernández-Urruzola, I.; Osma, N.; Packard, T. T.; Gómez, M.; Postel, L.

    2014-11-01

    The distribution of zooplankton biomass and potential metabolic rates, in terms of electron transport system (ETS) and glutamate dehydrogenase (GDH), were analyzed along a cross-shelf transect in waters off Namibia. The highly variable dynamics of upwelling filaments promoted short-term fluctuations in the zooplankton biomass and metabolism. Maximum values were characteristically found over the shelf-break, where zooplankton biomass as dry mass (DM) reached peaks of 64.5 mg m- 3 within the upper 200 m in late August. Two weeks later, the zooplankton-DM decreased by more than a third (19 mg DM m- 3). Zooplankton potential respiration and NH4+ excretion averaged 234 μmol O2 m- 3 d- 1 and 169 μmol NH4+ m- 3 d- 1 in the Namibian shelf, respectively. High protein-specific ETS activities even in the low-chlorophyll waters outside the filament suggested a shift into greater omnivory seaward. In this light, zooplankton elemental and isotopic compositions were used to investigate the pelagic food web interactions. They evidenced spatial changes in the carbon resource for zooplankton as well as changes in the form of nitrogen that fueled the biological production in aging advected waters. Overall, both aspects of zooplankton metabolism impacted the primary productivity at a level less than 10% under all the different oceanographic conditions.

  14. Coastal submarine hydrothermal activity of Northern Baja California 2. Evolutionary history and isotope geochemistry

    SciTech Connect

    Vidal, V.M.V.; Vidal, F.V.; Isaacs, J.D.

    1981-10-10

    A geochemical model of the Punta Banda submarine hydrothermal system (PBSHS) and Ensenada quadrangle subaerial hot springs is developed using /sup 18/O//sup 16/O, D/H, /sup 34/S//sup 32/S, /sup 3/H, water and gas chemistry. The PBSHS water is a primary high temperature, acid, reducing fluid of old seawater origin which has been titrated by cold, alkaline groundwater of meteoric origin. The final exiting solutions represent a 1:1 mixture of the two primary mixing components. In contrast, the subaerial hot spring waters are of unmixed meteoric origin. The subaerial hot spring gas is predominantly atmospheric N/sub 2/, while the PBSHS contains large amounts of CH/sub 4/ and N/sub 2/ derived from trapped marine sediments of Cretaceous age, deltaS/sup 34/ values of sampled hydrothermal waters are similar to Cretaceous marine sulfate values and suggest that the waters contacted Cretaceous marine sedimentary strata. The presence of the Alisitos and Rosario marine sedimentary formations of Cretaceous age within the Ensenada-Punta Banda quadrangel renders support to the above hypothesis. The data also demonstrate the pyrite mineralization and deposition in submarine hydrothermal environments result from the complexing of ferrous iron with elemental sulfur and sulfide and that submarine hydrothermal activity acts as a major source of silica, Ca/sup 2 +/, and trace metals and as a major sink for seawater Mg/sup 2 +/ and SO/sub 4//sup 2 -/.

  15. Chemical composition and antibacterial activity of Pinus halepensis Miller growing in West Northern of Algeria

    PubMed Central

    Fekih, Nadia; Allali, Hocine; Merghache, Salima; Chaïb, Faïza; Merghache, Djamila; El Amine, Mohamed; Djabou, Nassim; Muselli, Alain; Tabti, Boufeldja; Costa, Jean

    2014-01-01

    Objective To find new bioactive natural products, the chemical composition and to sudy the antibacterial activity of essential oil components extracted from the aerial parts of the Algerian aromatic plant Pinus halepensis Miller (P. halepensis) (needles, twigs and buds). Methods The essential oil used in this study was isolated by hydrodistillation using a Clevenger-type apparatus according to the European Pharmacopoeia. The chemical composition was investigated using GC-retention indices (RI) and GC-MS. Results Forty-nine compounds, representing 97.9% of the total collective oil, were identified. Essential oil was dominated by hydrocarbon compounds (80.6%) especially monoterpenes (65.5%). The major compounds from ten oils stations were: myrcene (15.2%-32.0%), α-pinene (12.2%-24.5%), E-β-caryophyllene (7.0%-17.1%), terpinolene (1.8%-13.3%), 2-phenyl ethyl isovalerate (4.8%-10.9%), terpinene-4-ol (1.0%-8.2 %) and sabinene (1.5%-6.3%). The intra-species variations of the chemical compositions of P. halepensis aerial parts essential oils from ten Algerian sample locations were investigated using statistical analysis. Essential oil samples were clustered in 2 groups by hierarchical cluster analysis, according to their chemical composition. The essential oil revealed an interesting antimicrobial effect against Lysteria monocytogenes, Enterococcus faecalis, Pseudomonas aeruginosa, Acinetobacter baumanii, Citrobacter freundii and Klebsiella pneumoniae. Conclusions These results suggest that the essential oil from P. halepensis may be a new potential source as natural antimicrobial applied in pharmaceutical and food industries.

  16. Signals of recent snow-avalanche activity in birch tree-rings from Northern Iceland colluvial cones

    NASA Astrophysics Data System (ADS)

    Decaulne, Armelle; Eggertsson, Ólafur; Sæmundsson, Şorsteinn; Páll Jónsson, Helgi

    2010-05-01

    The Fnjóskadalur, Ljósavatnskarð and Dalsmynni valleys, in Northern Iceland, are characterised by an important snow-avalanche activity impacting large colluvial cones. All valleys expose stands of broad-leaved trees, which cover an extensive wooded part along the flanks. The main represented species are Betula pubescens trees and shrubs. The objective of the research is to determine past snow-avalanche activity to improve historical record for the last century by applying dendrochronological and vegetative analysis. Trees and shrubs experience damages resulting from the impact of snow and debris, to which they respond in a variety of ways. The dendrochronological approach compares tree rings growth from a reference area beside the snow-avalanche path with the ones from within the snow-avalanche path. For this purpose, increment cores are taken from the up-down axis of the trunks and analysed on a LINTAB measuring table. The dendromorphological analysis maps changes in trunk posture such as tilted or topped trunks, and the position of wounds on the trunks. The combination of these two approaches provides a temporal catalogue of snow-avalanche events and also determines the directions of main fluxes. Therefore, it helps to locate the lateral dispersion of snow avalanches over the cones through time. The results obtained from comparison between the reference growth curve and the snow-avalanche impacted one show a clear difference between impeded tree-ring growth due to climatic factors and snow-avalanche occurrence. Several snow avalanche events are unravelled on the investigated cones during the last century.

  17. 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.

  18. Mitigating Radicalism in Northern Nigeria

    DTIC Science & Technology

    2013-08-01

    radicalization in northern Nigeria. u Active engagement of youth and communities in peacebuilding programs that facilitate interactions among individuals...leaders, sustained development investments in marginalized communities , promotion of values of inclusivity to mitigate the spread of extremist ideology...claiming to have repelled Boko Haram, the militants return, regroup, and seek revenge. As a result, social and economic activities in the northern

  19. 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.

  20. Northern cardiometeopathies.

    PubMed

    Hasnulin, V I; Sevost'yanova, E V; Hasnulina, A V

    2001-04-01

    Our research in high latitudes has allowed the identification of a special class of deadaptive disorders, northern cardiometeopathies, which integrates cardiovascular functional violations connected to biologically significant modifications of meteorological, geomagnetic, electrical, gravitational, rhythmological and other geo-ecological factors of the North. Cardiac and cerebral disorder complexes, and also some psychoemotional modifications manifest cardiometeopathies. Cardiometeopathies can occur with developing of pathology, and in such a case they can be selected in the independent form of deadaptive disease. At the same time, cardiometeopathies in case where cardiovascular pathology already exists could cause complications and become a particular risk factor for the development of injuries and myocardial heart attack. The most important mechanism of cardiometeopathies' formation is the organism's reaction to modifications of the Earth's electromagnetic field, based on internal electromagnetic fields' (first of all pulsing field of heart) dependence on the magnitude and directness of modification of the external electromagnetic fields. The analysis of the high degree of dependence of the blood circulation effiency during geomagnetic perturbations in the North of the modification of electromagnetic heart activity allows us to speak of the discovery of an earlier unknown electromagnetic blood pump.

  1. 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.

  2. Tracking of physical activity, fitness, body composition and diet from adolescence to young adulthood: The Young Hearts Project, Northern Ireland.

    PubMed

    Boreham, Colin; Robson, Paula J; Gallagher, Alison M; Cran, Gordon W; Savage, J Maurice; Murray, Liam J

    2004-10-05

    BACKGROUND: The assumption that lifestyles formed early in life track into adulthood has been used to justify the targeting of health promotion programmes towards children and adolescents. The aim of the current study was to use data from the Northern Ireland Young Hearts Project to ascertain the extent of tracking, between adolescence and young adulthood, of physical activity, aerobic fitness, selected anthropometric variables, and diet. METHODS: Males (n 245) and females (n 231) were assessed at age 15 y, and again in young adulthood [mean (SD) age 22 (1.6) y]. At both timepoints, height, weight and skinfold thicknesses were measured, and physical activity and diet were assessed by questionnaire and diet history method respectively. At 15y, fitness was assessed using the 20 metre shuttle run, while at young adulthood, the PWC170 cycle ergometer test was used. For each measurement made at 15y, subjects were ranked into 'low' (L1; lowest 25%), 'medium' (M1; middle 50%) or 'high' (H1; highest 25%) categories. At young adulthood, similar categories (L2, M2, H2) were created. The extent of tracking of each variable over time was calculated using 3 x 3 matrices constructed using these two sets of categories, and summarised using kappa (kappa) statistics. RESULTS: Tracking of diet and fitness was poor (kappa activity in males was fair (kappa 0.202), but was poor in females (kappa 0.021). In contrast, anthropometric variables such as weight, body mass index and sum of skinfolds tracked more strongly in females (kappa 0.540, kappa 0.307, kappa 0.357 respectively) than in males (kappa 0.337, kappa 0.199, kappa 0.216 respectively). CONCLUSIONS: The poor tracking of fitness and diet in both sexes, and physical activity in females, suggests that these aspects of adolescent lifestyle are unlikely to be predictive of behaviours in

  3. Hydrothermal activity along the northern Mid-Atlantic Ridge and in the Bransfield Strait Backarc Basin, Antarctica

    NASA Astrophysics Data System (ADS)

    Chin, Carol S.

    1998-12-01

    Seafloor hydrothermal circulation through young oceanic crust results in the expulsion of fluids as both diffuse and focused flow in the form of hydrothermal venting. High-temperature hydrothermal fluids are enriched in reduced chemical species that rapidly oxidize upon interaction with ambient, oxygen-rich bottom waters, resulting in plumes that are detectable in the water column both by their dissolved chemical composition as well as by their particle concentration. This study employed a novel instrument package which detected both dissolved manganese and particle concentration in situ. This package also included a standard CTD (conductivity, temperature, depth) and rosette for the collection of water samples. Because hydrothermal plumes integrate the output from an entire vent field, measurements in plumes can be used to estimate vent field fluxes. Some geochemical tracers from hydrothermal vents can also be detected thousands of kilometers from their sources. Thus, plumes provide the means to prospect for undiscovered hydrothermal sites, and can also predict characteristics of the venting site. This work includes studies of hydrothermal plumes along the northern Mid-Atlantic Ridge and in the Bransfield Strait backarc basin, Antarctica. In recent years, the number of known hydrothermal sites on the Mid-Atlantic Ridge (MAR) has increased from two to seven, and most other segments between 12° and 41° N have shown evidence of high-temperature hydrothermal activity. Furthermore, it appears that as one approaches the Azores Plateau, the concentration of dissolved delta3He in the bottom water (originating from hydrothermal venting) increases, suggesting that hydrothermal activity increases toward the plateau. This is consistent with the significant tectonic extension and crustal fissuring observed near the Azores Platform, which is expected to support increased convection. The Bransfield Strait is a backarc basin between the Antarctic Peninsula and the South

  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. 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.

  6. Seismological evidence of an active footwall shortcut thrust in the Northern Itoigawa-Shizuoka Tectonic Line derived by the aftershock sequence of the 2014 M 6.7 Northern Nagano earthquake

    NASA Astrophysics Data System (ADS)

    Panayotopoulos, Yannis; Hirata, Naoshi; Hashima, Akinori; Iwasaki, Takaya; Sakai, Shin'ichi; Sato, Hiroshi

    2016-06-01

    A destructive M 6.7 earthquake struck Northern Nagano prefecture on November 22, 2014. The main shock occurred on the Kamishiro fault segment of the northern Itoigawa-Shizuoka Tectonic Line (ISTL). We used data recorded at 41 stations of the local seismographic network in order to locate 2118 earthquakes that occurred between November 18 and November 30, 2014. To estimate hypocenters, we assigned low Vp models to stations within the Northern Fossa Magna (NFM) basin thus accounting for large lateral crustal heterogeneities across the Kamishiro fault. In order to further improve accuracy, the final hypocenter locations were recalculated inside a 3D velocity model using the double-difference method. We used the aftershock activity distribution and focal mechanism solutions of major events in order to estimate the source fault area of the main shock. Our analysis suggests that the shallow part of the source fault corresponds to the surface trace of the Kamishiro fault and dips 30°-45° SE, while the deeper part of the source fault corresponds to the downdip portion of the Otari-Nakayama fault, a high angle fault dipping 50°-65° SE that formed during the opening of the NFM basin in the Miocene. Along its surface trace the Otari-Nakayama fault has been inactive during the late Quaternary. We verified the validity of our model by calculating surface deformation using a simple homogeneous elastic half-space model and comparing it to observed surface deformation from satellite interferometry, assuming large coseismic slip in the areas of low seismicity and small coseismic slip in the areas of high seismicity. Shallowing of the source fault from 50°-65° to 30°-45° in the upper 4 km, in the areas where both surface fault traces are visible, is a result of footwall shortcut thrusting by the Kamishiro fault off the Otari-Nakayama fault.

  7. Analysis of Tissue Plasminogen Activator Eligibility by Sex in the Greater Cincinnati/Northern Kentucky Stroke Study

    PubMed Central

    Madsen, Tracy E.; Khoury, Jane C.; Alwell, Kathleen A.; Moomaw, Charles J.; Kissela, Brett M.; De Los Rios La Rosa, Felipe; Woo, Daniel; Adeoye, Opeolu; Flaherty, Matthew L.; Khatri, Pooja; Ferioli, Simona; Kleindorfer, Dawn

    2017-01-01

    Background and Purpose Sex differences in recombinant tissue-type plasminogen activator (r-tPA) administration are present in some populations. It is unknown whether this is because of eligibility differences or the modifiable exclusion criterion of severe hypertension. Our aim was to investigate sex differences in r-tPA eligibility, in individual exclusion criteria, and in the modifiable exclusion criterion, hypertension. Methods We included all ischemic stroke patients ≥18 years among residents of the Greater Cincinnati/Northern Kentucky region who presented to 16-area emergency departments in 2005. Eligibility for r-tPA and individual exclusion criteria were determined using 2013 American Heart Association (AHA) and European Cooperative Acute Stroke Study (ECASS) III guidelines. Results Of 1837 ischemic strokes, 58% were women, 24% were black. Mean age in years was 72.2 for women and 66.1 for men. Eligibility for r-tPA was similar by sex (6.8% men and 6.1% women; P=0.55), even after adjusting for age (7.0% and 5.9%; P=0.32). Similar proportions of women and men arrived beyond 3- and 4.5-hour time windows, but more women had severe hypertension. There were no sex differences in blood pressure treatment rates among those with severe hypertension (14.6% women and 20.8% men; P=0.21). More women were >80 years and had National Institutes of Health Stroke Scale (NIHSS) >25. Conclusions Within a large, biracial population, eligibility for r-tPA was similar by sex. Women were more likely to have the modifiable exclusion criterion of severe hypertension but were not more likely to be treated. Women were more likely to have 2 of the 5 ECASS III exclusion criteria. Undertreatment of hypertension in women is a potentially modifiable contributor to reported differences in r-tPA administration. PMID:25628307

  8. 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.

  9. 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.

  10. 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.

  11. 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

  12. 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.

  13. Evidence for enhanced debris flow activity in the Northern Calcareous Alps since the 1980s (Plansee, Austria)

    NASA Astrophysics Data System (ADS)

    Dietrich, Andreas; Krautblatter, Michael

    2016-04-01

    From 1950 to 2011 almost 80.000 people lost their lives through the occurrence of debris flow events (Dowling and Santi, 2014). Debris flows occur in all alpine regions due to intensive rainstorms and mobilisable loose debris. Due to their susceptible lithology, the Northern Calcareous Alps are affected by a double digit number of major hazard events per year. Some authors hypothesised a relation between an increasing frequency of heavy rainstorms and an increasing occurrence of landslides in general (Beniston and Douglas, 1996) and debris flows in special (Pelfini and Santilli, 2008), but yet there is only limited evidence. The Plansee catchment in the Ammergauer Alps consists of intensely jointed Upper Triassic Hauptdolomit lithology and therefore shows extreme debris flow activity. To investigate this activity in the last decades, the temporal and spatial development of eight active debris flow fans is examined with GIS and field mapping. The annual rates since the late 1940s are inferred accurately by using aerial photos from 1947, 1952, 1971, 1979, 1987, 2000 and 2010. These rates are compared to the mean Holocene/Lateglacial debris flow volume derived from the most prominent cone. The contact with the underlying till is revealed by electrical resistivity tomography (ERT). It shows that the mean annual debris flow volume has increased there by a factor of 10 from 1947-1952 (0.23 ± 0.07 10³m³/yr) to 1987-2000 (2.41 ± 0.66 10³m³/yr). A similar trend can be seen on all eight fans: mean post-1980 rates exceed pre-1980 rates by a factor of more than three. This increasing debris flow activity coincides with an enhanced rainstorm (def. 35 mm/d) frequency recorded at the nearest meteorological station. Since 1921 the frequency of heavy rainstorms has increased there on average by 10% per decade. Recent debris flow rates are also 2-3 times higher compared to mean Holocene/Lateglacial rates. Furthermore, we state a strong correlation between the non

  14. Relationships between soil heavy metal pollution and enzyme activities in mining areas of northern Hunan province, Central South China

    NASA Astrophysics Data System (ADS)

    Hu, Xue-Feng; Jiang, Ying; Shu, Ying

    2014-05-01

    Hunan province, Central South China, is a well-known nonferrous metal base in China. Mine exploiting and processing there, however, often lead to heavy metal pollution of farmland. To study the effects of mining activities on the soil environmental quality, four representative paddy fields, the HSG, SNJ, NT and THJ, in Y county, northern Hunan province, were investigated. It was found that the streams running through the HSG, SNJ and NT are severely contaminated due to the long-term discharge of untreated mineral wastewater from local indigenous mining factories. The stream at the HSG, for example, is brownish red in color, with high concentrations of Cu, Zn, Cd, Fe and Mn. The concentrations of Cu, Zn and Cd in all the stream water of the HSG, SNJ and NT exceed the maximum allowable levels of the Agricultural Irrigation Water Criteria of China. Correspondingly, the HSG, SNJ and NT are heavily polluted by Cu, Zn and Cd due to the long-term irrigation with the contaminated stream water. In comparison, both stream water and paddy fields of the THJ, far away from mining areas, are not contaminated by any heavy metals and hence regarded as a control in this study. The rice grain produced at the HSG, SNJ and NT has a high risk of Cd contamination. The rate of rice grain produced in the four paddy fields in Y county with Cd exceeding the safe level (Cd, 0.2 μg g-1) specified by the National Standards for Rice Quality and Safety of China reaches 90%. Cd content in the rice grain is positively significantly correlated with that in the paddy fields, especially with the content of diethylenetriaminepentaacetic acid (DTPA) - extracted Cd, suggesting that the heavy metal pollution of paddy fields has already posed a high risk to rice safety and human health. Soil enzyme activities and microbial biomass are significantly inhibited by the heavy metal pollution of the paddy fields. Microbial biomass C and N (MBC and MBN) at a severely contaminated site of the HSG are only 31

  15. 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.

  16. 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.

  17. 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.

  18. Northern Nutrition.

    ERIC Educational Resources Information Center

    Northwest Territories Dept. of Education, Yellowknife.

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

  19. 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.

  20. Mid to late Holocene aeolian activity revealed by a multiproxy peat record in continental CE Europe (Northern Romania)

    NASA Astrophysics Data System (ADS)

    Panait, Andrei Marian; Feurdean, Angelica; Hutchinson, Simon Mark; Tanţǎu, Ioan

    2016-04-01

    Peat bogs, and especially ombrogenous mire, are increasingly used as continental archives of aeolian dust and sand deposition. Since ombrogenous peat is formed above ground water level all the inputs are atmospheric. Dust is more influenced by regional climatic patterns due to its small size, whereas sand tends to record local patterns in storm frequency and intensity reflecting its larger particle size. However, both size fractions are significantly underused proxies of past climate variability. Here, an ombrogenous peat profile from Tǎul Muced in the Rodnei Mountains (Northern Romanian Carpathians), located in a temperate continental climate, with Atlantic and Baltic influences, provides the very first record of mid to late Holocene aeolian activity from Romania highlighting the interplay between local and regional controls in a continental area of CE Europe. We use a multiproxy approach combining radiocarbon dating, the physical properties of the peat (loss-on-ignition, bulk density), mineral magnetic measurements (ARM, SIRM), geochemical (Ti and Zr) and particle size analysis (via both laser diffraction and the manual counting of sand particles under a steromicroscope) to determine changes in: i) atmospheric dust deposition and ii) wind velocities during the last 7800 years. We found that the aeolian particles are mainly silt (3.9-63 μm) (dust) and sand (63-1200 μm). The mineralogical composition of the aeolian sediment in peat is mainly quartz, more rarely calcite and very rarely other minerals such as feldspar, sulphur, mica (biotite and muscovite), magnetite and other melanocrate minerals. The roundness of the sand particles varies from well-rounded to sub-angular and angular, and suggests that the sand particles have different source areas. Results from this study show that over the last 7600 years the pattern of wind frequency changed several times: there are periods characterised by a low aeolian input around 6950-6550, 5000-3900, 3500-2900, 1650

  1. Identifying buried segments of active faults in the northern Rio Grande Rift using aeromagnetic, LiDAR,and gravity data, south-central Colorado, USA

    USGS Publications Warehouse

    Ruleman, Cal; Grauch, V. J.

    2013-01-01

    Combined interpretation of aeromagnetic and LiDAR data builds on the strength of the aeromagnetic method to locate normal faults with significant offset under cover and the strength of LiDAR interpretation to identify the age and sense of motion of faults. Each data set helps resolve ambiguities in interpreting the other. In addition, gravity data can be used to infer the sense of motion for totally buried faults inferred solely from aeromagnetic data. Combined interpretation to identify active faults at the northern end of the San Luis Basin of the northern Rio Grande rift has confirmed general aspects of previous geologic mapping but has also provided significant improvements. The interpretation revises and extends mapped fault traces, confirms tectonic versus fluvial origins of steep stream banks, and gains additional information on the nature of active and potentially active partially and totally buried faults. Detailed morphology of surfaces mapped from the LiDAR data helps constrain ages of the faults that displace the deposits. The aeromagnetic data provide additional information about their extents in between discontinuous scarps and suggest that several totally buried, potentially active faults are present on both sides of the valley.

  2. 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.

  3. 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.

  4. 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

  5. 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...

  6. 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...

  7. 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

  8. 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…

  9. 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…

  10. 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...

  11. 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

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

    PubMed Central

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

    2016-01-01

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

  13. 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.

  14. 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.

  15. Glaciomarine sedimentation and bottom current activity on the north-western and northern continental margins of Svalbard during the late Quaternary

    NASA Astrophysics Data System (ADS)

    Chauhan, Teena; Noormets, Riko; Rasmussen, Tine L.

    2016-04-01

    Palaeo-bottom current strength of the West Spitsbergen Current (WSC) and the influence of the Svalbard-Barents Sea Ice Sheet (SBIS) on the depositional environment along the northern Svalbard margins are poorly known. Two gravity cores from the southern Yermak Plateau and the upper slope north of Nordaustlandet, covering marine isotope stage (MIS) 1 to MIS 5, are investigated. Five lithofacies, based on grain size distribution, silt/clay ratio, content and mean of sortable silt (SS), are distinguished to characterise the contourite-dominated sedimentary environments. In addition, depositional environments are described using total organic carbon (TOC), total sulphur (TS) and calcium carbonate (CaCO3) contents of sediments. Facies A, containing coarse SS, suggests strong bottom current activity and good bottom water ventilation conditions as inferred from low TOC content. This facies was deposited during the glacial periods MIS 4, MIS 2 and during the late Holocene. Facies B is dominated by fine SS indicating weak bottom current and poor ventilation (cf. high TOC content of 1.2-1.6%), and correlates with the MIS 4/3 and MIS 2/1 transition periods. With an equal amount of clay and sand, fine SS and high content of TOC, facies C indicates reduced bottom current strength for intervals with sediment supply from proximal sources such as icebergs, sea ice or meltwater discharge. This facies was deposited during the last glacial maximum. Facies D represents mass-flow deposits on the northern Svalbard margin attributed to the SBIS advance at or near the shelf edge. Facies E sediments indicating moderate bottom current strength were deposited during MIS 5 and MIS 3, and during parts of MIS 2. This first late Quaternary proxy record of the WSC flow and sedimentation history from the northern Svalbard margin suggests that the oceanographic conditions and ice sheet processes have exerted first-order control on sediment properties.

  16. 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.

  17. 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

  18. 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

  19. 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.

  20. A Comparative Analysis between GIMSS NDVIg and NDVI3g for Monitoring Vegetation Activity Change in the Northern Hemisphere during 1982-2008

    DOE PAGES

    Jiang, Nan; Zhu, Wenquan; Zheng, Zhoutao; ...

    2013-08-12

    The long-term Normalized Difference Vegetation Index (NDVI) time-series data set generated from the Advanced Very High Resolution Radiometers (AVHRR) has been widely used to monitor vegetation activity change. The third version of NDVI (NDVI3g) produced by the Global Inventory Modeling and Mapping Studies (GIMMS) group was released recently. The comparisons between the new and old versions should be conducted for linking existing studies with future applications of NDVI3g in monitoring vegetation activity change. Based on simple and piecewise linear regression methods, this research made a comparative analysis between NDVIg and NDVI3g for monitoring vegetation activity change and its responses tomore » climate change in the middle and high latitudes of the Northern Hemisphere during 1982–2008. Our results indicated that there were large differences between NDVIg and NDVI3g in the spatial patterns for both the overall changing trends and the timing of Turning Points (TP) in NDVI time series, which spread over almost the entire study region. The average NDVI trend from NDVI3g was almost twice as great as that from NDVIg and the detected average timing of TP from NDVI3g was about one year later. Although the general spatial patterns were consistent between two data sets for detecting the responses of growing-season NDVI to temperature and precipitation changes, there were large differences in the response magnitude, with a higher response magnitude to temperature in NDVI3g and an opposite response to precipitation change for the two data sets. Finally, these results demonstrated that the NDVIg data set may underestimate the vegetation activity change trend and its response to climate change in the middle and high latitudes of the Northern Hemisphere during the past three decades.« less

  1. A Comparative Analysis between GIMSS NDVIg and NDVI3g for Monitoring Vegetation Activity Change in the Northern Hemisphere during 1982-2008

    SciTech Connect

    Jiang, Nan; Zhu, Wenquan; Zheng, Zhoutao; Chen, Guangsheng; Fan, Deqin

    2013-08-12

    The long-term Normalized Difference Vegetation Index (NDVI) time-series data set generated from the Advanced Very High Resolution Radiometers (AVHRR) has been widely used to monitor vegetation activity change. The third version of NDVI (NDVI3g) produced by the Global Inventory Modeling and Mapping Studies (GIMMS) group was released recently. The comparisons between the new and old versions should be conducted for linking existing studies with future applications of NDVI3g in monitoring vegetation activity change. Based on simple and piecewise linear regression methods, this research made a comparative analysis between NDVIg and NDVI3g for monitoring vegetation activity change and its responses to climate change in the middle and high latitudes of the Northern Hemisphere during 1982–2008. Our results indicated that there were large differences between NDVIg and NDVI3g in the spatial patterns for both the overall changing trends and the timing of Turning Points (TP) in NDVI time series, which spread over almost the entire study region. The average NDVI trend from NDVI3g was almost twice as great as that from NDVIg and the detected average timing of TP from NDVI3g was about one year later. Although the general spatial patterns were consistent between two data sets for detecting the responses of growing-season NDVI to temperature and precipitation changes, there were large differences in the response magnitude, with a higher response magnitude to temperature in NDVI3g and an opposite response to precipitation change for the two data sets. Finally, these results demonstrated that the NDVIg data set may underestimate the vegetation activity change trend and its response to climate change in the middle and high latitudes of the Northern Hemisphere during the past three decades.

  2. 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)

  3. 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.

  4. 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

  5. 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.

  6. 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.

  7. 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.

  8. 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

  9. Active thrusting and folding along the northern Tien Shan and Late Cenozoic rotation of the Tarim relative to Dzungaria and Kazakhstan

    NASA Astrophysics Data System (ADS)

    Avouac, Jean-Philippe; Tapponnier, P.; Bai, M.; You, H.; Wang, G.

    1993-04-01

    Geometries and rates of Late Cenozoic thrust faulting and folding along the northern piedmont of the Tien Shan mountain belt, which was the location of the 1906 Manas earthquake, are studied. A flexural foredeep of the Dzungarian basement, filled with up to 11,000 m of sediment, is overthrusted by the 5,000 m northern range of the Tien Shan. The active thrust is found to reach the surface 30 km north of the front range, within a 200-km-long zone of Neogene-Quaternary anticlines. Cenozoic shortening of the folded Dzungarian sediments is estimated to be on the order of 30 km, and the Cenozoic shortening rate is found to have been 3.0 +/- 1.5 mm/yr. Reactivation of the Tien Shan by the collision of India and Asia is determined to have taken place in the early to middle Miocene. This is consistent with the existence of thick late Neogene and Quaternary deposits. Reactivation of the Tien Shan roughly coincides with the great mid-Miocene changes in tectonic regimes, denudation, and sedimentation rates seen in southeast Asia.

  10. 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

  11. A case control study of active genital Chlamydia trachomatis infection among patients with tubal infertility in northern Nigeria.

    PubMed

    Tukur, J; Shittu, S O; Abdul, A M

    2006-01-01

    A case-controlled study of the prevalence of genital Chlamydia trachomatis infection in 120 patients with tubal infertility (study group) and 120 clients of the family planning clinic (control group) attending the Ahmadu Bello University Teaching Hospital, Zaria in northern Nigeria is reported. The prevalence was 38.3%, 95% confidence interval (CI) (29.6-47.6%) in the study group and 13.3%, 95% CI (7.8-20.7%) in the control group. There was a fourfold risk of having genital Chlamydial infection among the cases compared to the controls (odds ratio [OR] = 4.04, 95% CI (2.04 < OR < 8.09). Tubal infertility was found to be significantly associated with genital C. trachomatis infection (P < 0.001). Out of the 58 cases of primary infertility, 22(37.9%) tested positive for genital C. trachomatis compared with 24 of the 62 (38.7%) that had secondary infertility. The infection was not found to be significantly associated with a particular type of infertility (primary or secondary), number of sexual partners or previous sexually transmitted disease (P < 0.05). There is need for provision of facilities to enable screening for genital C. trachomatis infection in view of its high prevalence in the study population.

  12. Incipient mantle delamination, active tectonics and crustal thickening in Northern Morocco: Insights from gravity data and numerical modeling

    NASA Astrophysics Data System (ADS)

    Baratin, Laura-May; Mazzotti, Stéphane; Chéry, Jean; Vernant, Philippe; Tahayt, Abdelilah; Mourabit, Taoufik

    2016-11-01

    The Betic-Rif orocline surrounding the Alboran Sea, the westernmost tip of the Mediterranean Sea, accommodates the NW-SE convergence between the Nubia and Eurasia plates. Recent GPS observations indicate a ∼4 mm/yr SW motion of the Rif Mountains, relative to stable Nubia, incompatible with a simple two-plate model. New gravity data acquired in this study define a pronounced negative Bouguer anomaly south of the Rif, interpreted as a ∼40 km-thick crust in a state of non-isostatic equilibrium. We study the correlation between these present-day kinematic and geodynamic processes using a finite-element code to model in 2-D the first-order behavior of a lithosphere affected by a downward normal traction (representing the pull of a high-density body in the upper mantle). We show that intermediate viscosities for the lower crust and uppermost mantle (1021-1022Pas) allow an efficient coupling between the mantle and the base of the brittle crust, thus enabling (1) the conversion of vertical movement, resulting from the downward traction, to horizontal movement and (2) shortening in the brittle upper crust. Our results show that incipient delamination of the Nubian continental lithosphere, linked to slab pull, can explain the present-day abnormal tectonics, contribute to the gravity anomaly observed in northern Morocco, and give insight into recent tectonics in the Western Mediterranean region.

  13. Long-term monthly statistics of mid-latitudinal NmF2 in the Northern geographic hemisphere during geomagnetically quiet and steadily low solar activity conditions

    NASA Astrophysics Data System (ADS)

    Pavlov, Anatoli; Pavlova, Nadezhda

    2016-07-01

    Long-term mid-latitude hourly values of NmF2 measured in 1957-2015 by 10 ionosondes (Point Arguello, Wallops Is., Boulder, de l'Ebre, Rome, Ottawa, Pruhonice, Dourbes, Slough, and Juliusruh) in the Northern geographic hemisphere were processed to select periods of geomagnetically quiet and low solar activity conditions to calculate several descriptive statistics of the noon NmF2 for each month, including the mathematical expectation, most probable value, arithmetic average, and arithmetic average median. The month-to-month variability of these descriptors allowed us to identify months of a year when they reach their extremes (maxima, minima). The calculated month-to-month variations of the NmF2 statistical parameters made it possible to study the winter anomaly and spring-autumn asymmetry in these statistical parameters.

  14. 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.

  15. 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.

  16. Maximal enzyme activities, and myoglobin and glutathione concentrations in heart, liver and skeletal muscle of the Northern Short-tailed shrew (Blarina brevicauda; Insectivora: Soricidae).

    PubMed

    Stewart, J M; Woods, A K; Blakely, J A

    2005-07-01

    We measured the enzymes of glycolysis, Krebs Cycle, beta-oxidation and electron transport in the heart, liver and skeletal muscle of the Northern Short-tailed Shrew, Blarina brevicauda. Additionally, we measured the amount of myoglobin in skeletal and heart muscle as well as the concentration of glutathione in heart. The picture that emerges is of an aerobically well-endowed animal with constrained anaerobic capacity as indicated by small activities of glycolytic enzymes and creatine kinase. Lipid metabolism and amino acid transamination, as well as gluconeogenesis, are predominant in processing carbon resources and probably reflect the large contribution lipid and protein make to the diet of this carnivore. The citrate synthase activity is the largest of any reported value for vertebrate heart (250 U/g). The additional, very active cytochrome c oxidase activity (220 U/g) and large myoglobin concentrations (8 mg/g) in heart are clearly the underpinnings of the rapid metabolic rates reported for small insectivores. The potential for generation of reactive oxygen species must be great since the total glutathione concentration (165 mumol/g) is 300-fold greater in shrew hearts than in hearts of rats.

  17. 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.

  18. 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.

  19. 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

  20. Northern Parkway PTA Makes Health a Habit

    ERIC Educational Resources Information Center

    Ferdinand, Marilyn

    2011-01-01

    Health and fitness have been on the agenda of Northern Parkway Elementary School for quite some time, thanks to the concerted efforts of its involved and active PTA officers and members. For the past five years, the Northern Parkway PTA has held a popular and well-attended Family Fun and Fitness Night and has complemented the activities and…

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. Seasonal and Diel Activity Patterns of Eight Sympatric Mammals in Northern Japan Revealed by an Intensive Camera-Trap Survey

    PubMed Central

    Ikeda, Takashi; Uchida, Kenta; Matsuura, Yukiko; Takahashi, Hiroshi; Yoshida, Tsuyoshi; Kaji, Koichi; Koizumi, Itsuro

    2016-01-01

    The activity patterns of mammals are generally categorized as nocturnal, diurnal, crepuscular (active at twilight), and cathemeral (active throughout the day). These patterns are highly variable across regions and seasons even within the same species. However, quantitative data is still lacking, particularly for sympatric species. We monitored the seasonal and diel activity patterns of terrestrial mammals in Hokkaido, Japan. Through an intensive camera-trap survey a total of 13,279 capture events were recorded from eight mammals over 20,344 camera-trap days, i.e., two years. Diel activity patterns were clearly divided into four categories: diurnal (Eurasian red squirrels), nocturnal (raccoon dogs and raccoons), crepuscular (sika deer and mountain hares), and cathemeral (Japanese martens, red foxes, and brown bears). Some crepuscular and cathemeral mammals shifted activity peaks across seasons. Particularly, sika deer changed peaks from twilight during spring–autumn to day-time in winter, possibly because of thermal constraints. Japanese martens were cathemeral during winter–summer, but nocturnal in autumn. We found no clear indication of predator-prey and competitive interactions, suggesting that animal densities are not very high or temporal niche partitioning is absent among the target species. This long-term camera-trap survey was highly cost-effective and provided one of the most detailed seasonal and diel activity patterns in multiple sympatric mammals under natural conditions. PMID:27732611

  8. Activation of systemic, but not local, renin-angiotensin system is associated with upregulation of TNF-α during prolonged fasting in northern elephant seal pups.

    PubMed

    Suzuki, Miwa; Vázquez-Medina, José Pablo; Viscarra, Jose A; Soñanez-Organis, José G; Crocker, Daniel E; Ortiz, Rudy M

    2013-09-01

    Northern elephant seal pups naturally endure a 2-3 month post-weaning fast that is associated with activation of systemic renin-angiotensin system (RAS), a decrease in plasma adiponectin (Acrp30), and insulin resistance (IR)-like conditions. Angiotensin II (Ang II) and tumor necrosis factor-alpha (TNF-α) are potential causal factors of IR, while Acrp30 may improve insulin signaling. However, the effects of fasting-induced activation of RAS on IR-like conditions in seals are not well described. To assess the effects of prolonged food deprivation on systemic and local RAS, and their potential contribution to TNF-α as they relate to an IR condition, the mRNA expressions of adipose and muscle RAS components and immuno-relevant molecules were measured along with plasma RAS components. Mean plasma renin activity and Ang II concentrations increased by 89 and 1658%, respectively, while plasma angiotensinogen (AGT) decreased by 49% over the fast, indicative of systemic RAS activation. Prolonged fasting was associated with decreases in adipose and muscle AGT mRNA expressions of 69 and 68%, respectively, corresponding with decreases in tissue protein content, suggesting suppression of local AGT production. Muscle TNF-α mRNA and protein increased by 239 and 314%, whereas those of adipose Acrp30 decreased by 32 and 98%, respectively. Collectively, this study suggests that prolonged fasting activates a systemic RAS, which contributes to an increase in muscle TNF-α and suppression of adipose Acrp30. This targeted and tissue-specific regulation of TNF-α and Acrp30 is likely coordinated to synergistically contribute to the development of an IR-like condition, independent of local RAS activity. These data enhance our understanding of the adaptive mechanisms evolved by elephant seals to tolerate potentially detrimental conditions.

  9. Culturable bacterial flora associated with the dinoflagellate green Noctiluca miliaris during active and declining bloom phases in the Northern Arabian Sea.

    PubMed

    Basu, Subhajit; Deobagkar, Deepti D; Matondkar, S G Prabhu; Furtado, Irene

    2013-05-01

    A massive algal bloom of the dinoflagellate Noctiluca miliaris (green) was located in the Northern Arabian Sea by IRS-P4-2 (OCM-II) for microbiological studies, during two consecutive cruises of February-March 2009. Culturable bacterial load during bloom were ≈ 2-3-fold higher in comparison to non-bloom waters and ranged from 3.20 × 10(5) to 6.84 × 10(5) cfu ml(-1). An analysis of the dominant heterotrophs associated with Noctiluca bloom resulted in phylogenetic and a detailed metabolic characterization of 70 bacterial isolates from an overlapping active and declining bloom phase location near north-central Arabian Sea. The active phase flora was dominated by Gram-positive forms (70.59 %), a majority of which belonged to Bacillus (35.29 %) of Firmicutes. As the bloom declined, Gram-negative forms (61.11 %) emerged dominant, and these belonged to a diverse γ-proteobacterial population consisting of Shewanella (16.67 %) and equal fractions of a Cobetia-Pseudomonas-Psychrobacter-Halomonas population (36.11 %). A Unifrac-based principal coordinate analysis of partial 16S rDNA sequences showed significant differences among the active and declining phase flora and also with reported endocytic flora of Noctiluca (red). A nonparametric multidimensional scaling (NMDS) of antibiogram helped differentiation among closely related strains. The organic matter synthesized by N. miliaris appears to be quickly utilized and remineralized as seen from the high efficiency of isolates to metabolize various complex and simple C/N substrates such as carbohydrates, proteins/amino acids, lipids, sulfide production from organic matter, and solubilize phosphates. The ability of a large fraction of these strains (50-41.67 %) to further aerobically denitrify indicates their potential for nitrogen removal from these high-organic microniches of the Noctiluca bloom in the Arabian Sea, also known for high denitrification activity. The results indicate that culturable euphotic bacterial

  10. Prolonged fasting activates hypoxia inducible factors-1α, -2α and -3α in a tissue-specific manner in northern elephant seal pups.

    PubMed

    Soñanez-Organis, José G; Vázquez-Medina, José P; Crocker, Daniel E; Ortiz, Rudy M

    2013-09-10

    Hypoxia inducible factors (HIFs) are important regulators of energy homeostasis and cellular adaptation to low oxygen conditions. Northern elephant seals are naturally adapted to prolonged periods (1-2 months) of food deprivation (fasting) which result in metabolic changes that may activate HIF-1. However, the effects of prolonged fasting on HIFs are not well defined. We obtained the full-length cDNAs of HIF-1α and HIF-2α, and partial cDNA of HIF-3α in northern elephant seal pups. We also measured mRNA and nuclear protein content of HIF-1α, -2α, -3α in muscle and adipose during prolonged fasting (1, 3, 5 & 7 weeks), along with mRNA expression of HIF-mediated genes, LDH and VEGF. HIF-1α, -2α and -3α are 2595, 2852 and 1842 bp and encode proteins of 823, 864 and 586 amino acid residues with conserved domains needed for their function (bHLH and PAS) and regulation (ODD and TAD). HIF-1α and -2α mRNA expression increased 3- to 5-fold after 7 weeks of fasting in adipose and muscle, whereas HIF-3α increased 5-fold after 7 weeks of fasting in adipose. HIF-2α protein expression was detected in nuclear fractions from adipose and muscle, increasing approximately 2-fold, respectively with fasting. Expression of VEGF increased 3-fold after 7 weeks in adipose and muscle, whereas LDH mRNA expression increased 12-fold after 7 weeks in adipose. While the 3 HIFα genes are expressed in muscle and adipose, only HIF-2α protein was detectable in the nucleus suggesting that HIF-2α may contribute more significantly in the up-regulation of genes involved in the metabolic adaptation during fasting in the elephant seal.

  11. 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.

  12. 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

  13. 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

  14. Examining Activity-Based Learning (ABL) Practices in Public Basic Schools in the Northern Region of Ghana

    ERIC Educational Resources Information Center

    Nudzor, Hope P.; Dare, Albert; Oduro, George K. T.; Bosu, Rosemary; Addy, Nii

    2015-01-01

    Background: Ghana has been the testing ground for many teaching and learning initiatives over the past 15-20 years. These initiatives, largely funded by donors, have sought to improve learning by introducing and reinforcing valuable teaching skills, materials and approaches, most of them child-friendly, learner-centred and involving activity-based…

  15. 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

  16. 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.

  17. 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.

  18. 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.

  19. Antimicrobial activity and agricultural properties of bitter melon (Momordica charantia L.) grown in northern parts of Turkey: a case study for adaptation.

    PubMed

    Yaldız, Gülsüm; Sekeroglu, Nazım; Kulak, Muhittin; Demirkol, Gürkan

    2015-01-01

    This study was designed to determine the adaptation capability of bitter melon (Momordica charantia L.), which is widely grown in tropical and subtropical climates, in northern parts of Turkey. In this study, plant height, number of fruits, fruit length, fruit width, number of seeds and fruit weight of bitter melon grown in field conditions were determined. The antimicrobial effect of the ethanol extract of fruit and seeds against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus, Salmonella typhi, Aspergillus niger and Candida albicans microorganisms was tested in vitro by the disc diffusion method. In conclusion, plant height (260 cm), number of fruits (16 per  plant), number of seeds (30.2  per fruit), fruit width (3.8 cm), fruit length (10.6 cm) and fruit weight (117.28 g fruit(- 1)) were determined; fruits were found to have antimicrobial activity against A. niger; oil and seeds were found to have antimicrobial activity against A. niger and E. coli.

  20. Gender Differences in Health Related Quality of Life among People Living with HIV on Highly Active Antiretroviral Therapy in Mekelle Town, Northern Ethiopia

    PubMed Central

    Tesfay, Amanuel; Gebremariam, Abebe; Abrha, Hailay

    2015-01-01

    Background. Health related quality of life (HRQOL) is an important outcome measure for highly active antiretroviral treatment program. In Ethiopia, studies revealed that there are improved qualities of life among adults living with the viruses taking antiretroviral therapy but there is no explicit data showing gender differences in health related quality of life. Aim. To assess gender differences in HRQOL and its associated factors among people living with HIV and on highly active antiretroviral therapy in public health institutions of Mekelle town, Northern Ethiopia. Methods. A comparative cross-sectional study was conducted among 494 adult people living with HIV taking ART services. Quality of life was measured using WHOQOL-HIV BREF. Result. There was a statistically significant gender difference (P < 0.05) in HRQOL among PLHIV on HAART. Females had low score in all HRQOL domains. High perceived stigma was strongly associated with poor psychological quality of domain among both female and male groups with [AOR = 2.89(1.69,4.96)] and [AOR = 2.5(1.4,4.4)], respectively. Conclusion. There was statistically significant gender difference in all quality of life domains. Public health interventions to improve HRQOL of PLHIV should take in to account the physical, psychological, social, environmental, and spiritual health of PLHIV during treatment, care, and support. PMID:25632393

  1. 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

  2. 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.

  3. 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

  4. 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.

  5. 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.

  6. 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.

  7. Kinematic analysis of recent and active faults of the southern Umbria-Marche domain, Northern Apennines, Italy: geological constraints to geodynamic models

    NASA Astrophysics Data System (ADS)

    Pasqui, Valeria; Viti, Marcello; Mantovani, Enzo

    2013-04-01

    The recent and active deformation that affects the crest zone of the Umbria-Marche belt (Northern Apennines, Italy) displays a remarkable extensional character, outlined by development of normal fault sets that overprint pre-existing folds and thrusts of Late Miocene-Early Pliocene age. The main extensional fault systems often bound intermontane depressions hosting recent, mainly continental, i.e. fluvial or lacustrine deposits, separating the latter from Triassic-Miocene, mainly carbonatic and siliciclastic marine rocks that belong to the Romagna-Umbria-Marche stratigraphic succession. Stratigraphic data indicate that the extensional strain responsible for the development of normal fault-bounded continental basins in the outer zones of the Northern Apennines was active until Middle Pleistocene time. Since Middle Pleistocene time onwards a major geodynamic change has affected the Central Mediterranean region, with local reorganization of the kinematics in the Adria domain and adjacent Apennine belt. A wide literature illustrates that the overall deformation field of the Central Mediterranean area is presently governed by the relative movements between the Eurasia and Africa plates. The complex interaction of the Africa-Adria and the Anatolian-Aegean-Balkan domains has led the Adria microplate to migrate NW-ward and to collide against Eurasia along the Eastern Southern Alps. As a consequence Adria is presently moving with a general left-lateral displacement with respect to the Apennine mountain belt. The sinistral component of active deformations is also supported by analysis of earthquake focal mechanisms. A comparison between geophysical and geological evidence outlines an apparent discrepancy: most recognized recent and active faults display a remarkable extensional character, as shown by the geometry of continental basin-bounding structutes, whereas geodetic and seismologic evidence indicates the persistency of an active strike-slip, left-lateral dominated

  8. Climate change in winter versus the growing-season leads to different effects on soil microbial activity in northern hardwood forests

    NASA Astrophysics Data System (ADS)

    Sorensen, P. O.; Templer, P. H.; Finzi, A.

    2014-12-01

    Mean winter air temperatures have risen by approximately 2.5˚ C per decade over the last fifty years in the northeastern U.S., reducing the maximum depth of winter snowpack by approximately 26 cm over this period and the duration of winter snow cover by 3.6 to 4.2 days per decade. Forest soils in this region are projected to experience a greater number of freeze-thaw cycles and lower minimum winter soil temperatures as the depth and duration of winter snow cover declines in the next century. Climate change is likely to result not only in lower soil temperatures during winter, but also higher soil temperatures during the growing-season. We conducted two complementary experiments to determine how colder soils in winter and warmer soils in the growing-season affect microbial activity in hardwood forests at Harvard Forest, MA and Hubbard Brook Experimental Forest, NH. A combination of removing snow via shoveling and buried heating cables were used to induce freeze-thaw events during winter and to warm soils 5˚C above ambient temperatures during the growing-season. Increasing the depth and duration of soil frost via snow-removal resulted in short-term reductions in soil nitrogen (N) production via microbial proteolytic enzyme activity and net N mineralization following snowmelt, prior to tree leaf-out. Declining mass specific rates of carbon (C) and N mineralization associated with five years of snow removal at Hubbard Brook Experimental Forest may be an indication of microbial physiological adaptation to winter climate change. Freeze-thaw cycles during winter reduced microbial extracellular enzyme activity and the temperature sensitivity of microbial C and N mineralization during the growing-season, potentially offsetting nutrient and soil C losses due to soil warming in the growing-season. Our multiple experimental approaches show that winter climate change is likely to contribute to reduced microbial activity in northern hardwood forests.

  9. Insecticide susceptibility status and major detoxifying enzymes' activity in Aedes albopictus (Skuse), vector of dengue and chikungunya in Northern part of West Bengal, India.

    PubMed

    Bharati, Minu; Saha, Dhiraj

    2017-02-27

    Mosquitoes belonging to Aedes genus, Aedes aegypti and Aedes albopictus transmit many globally important arboviruses including Dengue (DENV) and Chikungunya (CHIKV). Vector control with the use of insecticide remains the suitable method of choice to stop the transmission of these diseases. However, vector control throughout the world is failing to achieve its target results because of the worldwide development of insecticide resistance in mosquitoes. To assess the insecticide susceptibility status of Aedes albopictus from northern part of West Bengal, the susceptibility of eight different Aedes albopictus populations were tested against a commonly used larvicide (temephos) and some adulticides (malathion, deltamethrin and lambda cyhalothrin) along with the major insecticide detoxifying enzymes' activity in them. Through this study, it was revealed that most of the populations were found susceptible to temephos except Nagrakata (NGK) and Siliguri (SLG), which showed both a higher resistance ratio (RR99) and a lower susceptibility, thereby reflecting the development of resistance against temephos in them. However, all tested adulticides caused 100% mortality in all the population implying their potency in control of this mosquito in this region of India. Through the study of carboxylesterase activity, it was revealed that the NGK population showed a 9.6 fold higher level of activity than susceptible population. The same population also showed a lower level of susceptibility and a higher resistance ratio (RR99), indicating a clear correlation between susceptibility to temephos and carboxylesterase enzymes' activity in this population. This preliminary data reflects that the NGK population is showing a trend towards resistance development and with time, there is possibility that this resistance phenomenon will spread to other populations. With the recurrence of dengue and chikungunya, this data on insecticide susceptibility status of Aedes albopictus could help the

  10. Physical Activity and Fitness of First Nations Youth in a Remote and Isolated Northern Ontario Community: A Needs Assessment.

    PubMed

    Gates, Michelle; Hanning, Rhona; Gates, Allison; Stephen, Judy; Fehst, Andrew; Tsuji, Leonard

    2016-02-01

    Among a group of First Nations youth, this research aimed to obtain objective measures of anthropometry, physical activity (PA) and fitness; to identify any group-level differences by sex, body mass index, waist circumference and body fat categories; to assess the barriers and supports to PA. Youth participated in anthropometric measures (BMI, waist circumference, body fat percentage), PA assessment (3 days of accelerometry) and fitness testing (guided by the Canadian Physical Activity, Fitness and Lifestyle Approach). Barriers and supports were assessed via environmental scan and focus groups. Descriptive statistics were compared to reference data. Group differences by sex, BMI status, waist circumference and body fat categories were tested using Mann-Whitney U and Chi square tests (p ≤ 0.05). Qualitative data were assembled into one file and coded manually for categories and themes. Seventy-two youth (12.1 ± 1.1 years, 61.1% male) participated in at least one measure; 36 completed the accelerometry. Sixty-three percent were overweight or obese, 51% were abdominally obese and 21% had excess body fat. Most (86.1%) met Canada's PA guidelines. Boys were more active than girls (p = 0.025) and had greater cardiorespiratory endurance (p = 0.003). Overweight, obese, or abdominally obese youth had lower cardiorespiratory endurance than normal weight youth (p < 0.001). Barriers and supports fell under the main themes: motivation, role models, personnel and facilities, environment and programs. Based on this assessment, youth in this community are active, but not sufficiently physically fit, especially among those affected by obesity and abdominal obesity. The findings, in addition to the numerous barriers to PA, support the community's desire for school-based PA programming.

  11. HYDRATE RESEARCH ACTIVITIES THAT BOTH SUPPORT AND DERIVE FROM THE MONITORING STATION/SEA-FLOOR OBSERVATORY, MISSISSIPPI CANYON 118, NORTHERN GULF OF MEXICO

    SciTech Connect

    Lutken, Carol

    2013-07-31

    A permanent observatory has been installed on the seafloor at Federal Lease Block, Mississippi Canyon 118 (MC118), northern Gulf of Mexico. Researched and designed by the Gulf of Mexico Hydrates Research Consortium (GOM-HRC) with the geological, geophysical, geochemical and biological characterization of in situ gas hydrates systems as the research goal, the site has been designated by the Bureau of Ocean Energy Management as a permanent Research Reserve where studies of hydrates and related ocean systems may take place continuously and cooperatively into the foreseeable future. The predominant seafloor feature at MC118 is a carbonate-hydrate complex, officially named Woolsey Mound for the founder of both the GOM-HRC and the concept of the permanent seafloor hydrates research facility, the late James Robert “Bob” Woolsey. As primary investigator of the overall project until his death in mid-2008, Woolsey provided key scientific input and served as chief administrator for the Monitoring Station/ Seafloor Observatory (MS-SFO). This final technical report presents highlights of research and accomplishments to date. Although not all projects reached the status originally envisioned, they are all either complete or positioned for completion at the earliest opportunity. All Department of Energy funds have been exhausted in this effort but, in addition, leveraged to great advantage with additional federal input to the project and matched efforts and resources. This report contains final reports on all subcontracts issued by the University of Mississippi, Administrators of the project, Hydrate research activities that both support and derive from the monitoring station/sea-floor Observatory, Mississippi Canyon 118, northern Gulf of Mexico, as well as status reports on the major components of the project. All subcontractors have fulfilled their primary obligations. Without continued funds designated for further project development, the Monitoring Station

  12. Developing a Small-scale De-fluoridation Filter for use in Rural Northern Ghana with Activated Alumina as the Sorbent

    NASA Astrophysics Data System (ADS)

    Craig, L.; Stillings, L. L.; Decker, D.; Thomas, J.

    2013-12-01

    In northern Ghana, groundwater is the main source of household water and is generally considered a safe and economical source of drinking water. However in some areas it contains fluoride (F-) concentrations above the 1.5 ppm limit recommended by the World Health Organization, putting the users at risk of fluorosis. The study area in the Upper East Region of northern Ghana has pockets of groundwater F- up to 4.6 ppm and, as a result, also has a high percentage of residents with dental fluorosis. They have no alternative water source and, because of the poverty and limited access to technology, the affected community lacks the capacity to set up advanced treatment systems. One proposed solution is to attach F- adsorption filters to the wells, since adsorption is considered a simple and cost effective approach for treating high F- drinking water. This study evaluates activated alumina as a sorbent for use in de-fluoridation filters in the study area. We evaluated the long-term adsorption capacity of activated alumina, as well as potential changes in F- adsorption rate and capacity with grain size. We measured differences in positive surface charge (as C m-2) via slow acid titration, as well as F- loading with varied prior hydration time. Experimental results from this research show no notable change in F- adsorption or positive surface charge when the activated alumina surface was pre-equilibrated in distilled water from 24 hours up to 30 weeks before the experiment. The results of F- loading show a maximum of ~3.4 mg F- sorbed per gm activated alumina (at initial pH ~6.9, initial F- 1 to 60 ppm, and 20 hr reaction time). The pH dependent surface charge shows a maximum of ~0.14 C m-2 at pH of ~4.4 and zero surface charge at pH ~8.5. F- loading experiments were conducted with grain size ranges 0.125 to 0.250 mm and 0.5 to 1.0 mm to evaluate changes in F- adsorption rate (initial pH ~6.9, initial F- 10 ppm) and F- loading (initial pH ~6.9, initial F- 1 to 60 ppm, 20 hr

  13. Authigenic carbonates from an active cold seep of the northern South China Sea: New insights into fluid sources and past seepage activity

    NASA Astrophysics Data System (ADS)

    Feng, Dong; Chen, Duofu

    2015-12-01

    Site F (also named Formosa ridge) represents the most vigorous cold seep on the northern South China Sea continental slope. In order to constrain the fluid sources and intensities of seepage, we investigated the petrography, mineralogy, stable carbon and oxygen isotopic compositions, element geochemistry and radiocarbon dating of authigenic carbonate rocks retrieved from the seafloor. Carbonate rocks mainly occurred as crusts, nodules, and nodular masses incorporated in carbonate breccias. The carbonates were comprised mainly of high-Mg calcite and aragonite. The δ13C of authigenic carbonate varied from -55.3‰ to -34.3‰ (mean: -48.5‰; n=47) vs. V-PDB, suggesting biogenic methane is the dominant carbon source fuelling the system. The δ18OCarbonate values were from +3.6‰ to +4.8‰ (mean: +3.9‰; n=47). The observed 18O-enrichement in relation to calculated equilibrium values in the carbonates probably reflects dissolution of gas hydrates. Combination of seafloor observations and the obtained AMS 14C ages suggest that (1) initiation of methane seepage from at least 10.6 ka ago; (2) environmental conditions may have been favorable for enhanced fluid seepage around 6 ka BP and (3) relatively low intensity of seepage from 2 ka BP till today.

  14. Variability of CCN Activation Behaviour of Aerosol Particles in the Marine Boundary Layer of the Northern and Southern Atlantic Ocean

    NASA Astrophysics Data System (ADS)

    Henning, Silvia; Dieckmann, Katrin; Hartmann, Susan; Schäfer, Michael; Wu, Zhijun; Merkel, Maik; Wiedensohler, Alfred; Stratmann, Frank

    2013-04-01

    The variability of cloud condensation nucleus (CCN) activation behaviour and total CCN number concentrations was investigated during three ship cruises. Measurements were performed in a mobile laboratory on the German research vessel FS Polarstern cruising between Cape Town and Bremerhaven (April / May and October / November 2011) as well as between Punta Arenas and Bremerhaven (April / May 2012). CCN size distributions were measured for supersaturations between 0.1% and 0.4% using a Cloud Condensation Nucleus Counter (DMT, USA). Aerosol particle and CCN total number concentrations as well as the hygroscopicity parameter κ (Petters and Kreidenweis, 2007) were determined. Furthermore, size distribution data were collected. The hygroscopicity parameter κ featured a high variability during the cruises, with a median κ-value of 0.52 ± 0.26. The κ-values are depended on air mass origin; and are as expected mainly dominated by marine influences, but also long range transport of aerosol particles was detected. In the Celtic Sea, κ was found to be lower than that of clean marine aerosol particles (0.72 ± 0.24; Pringle et al., 2010) with κ-values ~0.2, possibly influenced by anthropogenic emissions from Europe. Close to the West African coast particle hygroscopicity was found to be influenced by the Saharan dust plume, resulting in low κ-values ~0.25. Petters, M.D. and S.M. Kreidenweis (2007), A single parameter representation of hygroscopic growth and cloud condensation nucleus activity, Atmos. Chem. and Phys., 7, 1961-1971. Pringle, K.J., H. Tost, A. Pozzer, U. Pöschl, and J. Lelieveld (2010), Global distribution of the effective aerosol hygroscopicity parameter for CCN activation, Atmos. Chem. Phys., 10, 5241-5255.

  15. The response of nitrifying microbial assemblages to ammonium (NH4+) enrichment from salmon farm activities in a northern Chilean Fjord

    NASA Astrophysics Data System (ADS)

    Elizondo-Patrone, Claudia; Hernández, Klaudia; Yannicelli, Beatriz; Olsen, Lasse Mork; Molina, Verónica

    2015-12-01

    The consequences of aquaculture include alterations in nitrogen cycling in aquatic environments that may lead to ecosystem degradation. Herein salmon aquaculture release of ammonium (NH4+) to the water column and its effects on natural archaea and bacteria ammonia-oxidizers (AOA and AOB) and nitrite-oxidizing bacteria (NOB) community structure were studied in the Comau fjord using molecular approaches, such as: cloning (AOA and AOB richness), qPCR for C. Nitrosopumilus maritimus (AOA) and Nitrospina sp. (NOB) abundance (DNA) and RT-qPCR only for Nitrospina sp activity (RNA). Sampling was carried out in brackish (0.7-25 salinity, <5 m depth) and marine (>30 salinity, 25 m depth) waters during contrasting salmon production periods: rest (winter 2012), growth and harvest (summer and winter 2013). During the rest period, the highest NH4+ concentration was observed at Vodudahue River, whereas during productive periods NH4+ accumulated in the brackish layer inside salmon cages and in the vicinty (up to 700 m distance from the cages). The nitrifier community from the fjord reference station (Stn-C) was characterized by C. N. maritimus (AOA) and Nitrosomonas sp. (AOB) sequences affiliated with cosmopolitan ecotypes (e.g., marine, freshwater, hydrothermal), maxima abundances of C. N. maritimus (AOA) and Nitrospina sp. and extreme ranges of Nitrospina sp. activity occurred in the brackish layer. During productive periods, abundances of C. N. maritimus were co-varied with NH4+ concentrations inside salmon cages (summer) and the adjacent areas (winter). Productive periods were characterized by lower abundances but more homogeneity between brackish and marine areas than for the Stn-C nitrifiers. The physiological state of Nitrospina sp. estimated from cDNA:DNA ratios indicated higher growth during winter 2013 associated with NH4+ enrichment derived from production and river input. Our results suggest that in Comau Fjord, NH4+ enrichment events occur during salmon production and

  16. Late cretaceous extensional tectonics and associated igneous activity on the northern margin of the Gulf of Mexico Basin

    NASA Technical Reports Server (NTRS)

    Bowen, R. L.; Sundeen, D. A.

    1985-01-01

    Major, dominantly compressional, orogenic episodes (Taconic, Acadian, Alleghenian) affected eastern North America during the Paleozoic. During the Mesozoic, in contrast, this same region was principally affected by epeirogenic and extensional tectonism; one episode of comparatively more intense tectonic activity involving extensive faulting, uplift, sedimentation, intrusion and effusion produced the Newark Series of eposits and fault block phenomena. This event, termed the Palisades Disturbance, took place during the Late Triassic - Earliest Jurassic. The authors document a comparable extensional tectonic-igneous event occurring during the Late Cretaceous (Early Gulfian; Cenomanian-Santonian) along the southern margin of the cratonic platform from Arkansas to Georgia.

  17. Detection of one single mutation predicts thiopurine S-methyltransferase activity in a population of Saami in northern Norway.

    PubMed

    Loennechen, T; Utsi, E; Hartz, I; Lysaa, R; Kildalsen, H; Aarbakke, J

    2001-08-01

    Thiopurine S-methyltransferase (TPMT) activity exhibits genetic polymorphism. The purpose of this investigation was to identify TPMT mutant alleles in the Saami population as a basis of developing genotyping tests for prediction of TPMT activity. The most predominant allele in Saamis (n = 194) was the TPMT*3C allele (A719G mutation) representing 92% of the mutant alleles, with an estimated allelic frequency of 3.3%. The most frequent allele in Caucasians (n = 66) living in the same geographic area was the TPMT*3A (A719G and G460A mutations) representing 91% of the mutant alleles, with an estimated allelic frequency of 3.4%. A test for one mutation, A719G, may prospectively identify more than 90% of the Saami individuals who require reduction in thiopurine dose to avoid hematopoietic toxicity. In a Norwegian population, comprising both the major Caucasian population and a minor Saami population, the same genotyping tests (eg, tests for the A719G and G460A mutations) may be used.

  18. Meeting Northern Arizona's Supported Employment Training Needs.

    ERIC Educational Resources Information Center

    Martin, William E., Jr.; And Others

    In 1989 Northern Arizona University established a Supported Employment Training Center (SETC) to increase the number of trained job coaches in northern Arizona and provide knowledge and skills in supported employment to personnel from cooperating schools and agencies. First-year SETC activities focused on assessment of the training needs of…

  19. Black Carbon, Metal Concentrations and Lead Isotopes Ratios in Aerosols as Tracers of Human and Natural Activities in Northern Vietnam

    NASA Astrophysics Data System (ADS)

    Guinot, B. P.

    2015-12-01

    Atmospheric brown clouds (ABC) observed as widespread layers of brownish haze are regional scale plumes of air pollutants with a hot spot of emission located in East Asia. ABC are mainly composed of aerosol particles such as Black Carbon (BC) emitted to the atmosphere during biomass burning and fossil fuels combustion. The atmospheric lifetime of BC ranges from a few days in wet season up to one month in dry season. The use of stable lead isotopes and 21 elements as tracers of air pollution was applied to identify and characterized the main sources of anthropogenic activities in Asian region. Aerosol samples from Haiphong (North Vietnam) were collected by a high volume sampler for a period of one year from October 2012 to October 2013. Vietnam's 207Pb/206Pb ratios were almost identical to those found for China. Ratios of 207Pb/206Pb ranged from 0.837 to 0.871 which agrees with values previously reported for the last 10 years in China (0.841 - 0.879). No significant variation in isotope ratio was observed during the sampling period, which suggests that there was no large seasonal variation in the isotope ratios of airborne lead. Trajectory analysis showed that almost two third of the air masses originated from East Northeast which implies that China was a major source of lead in atmosphere. Enrichment factor calculations indicated a large influence of coal activity (EF(Al) As = 1982 ± 796, EF(Al) Cd = 972 ± 659, EF(Al) Sb = 1358 ± 930) but the difference between combustion and mining exploitation could not be evidenced. Significant correlations were found between two others groups of elements: As, Cu, Ni, Zn, and Al, Fe K, Co. Wind dilution was effective on metals concentration variation. During the cold and dry season (winter) ambient concentrations were high and variable, during the warm and wet season (summer) concentrations were stable and low. Taken together, these factors also identified industrial and lithogenic activities in the region.

  20. A detection method of subrecent to recent tectonic activity in the anticlinal system of the northern Negev, Israel

    SciTech Connect

    Zilberman, E.; Wachs, D. )

    1988-02-01

    Geomorphological and geophysical methods combined with borehole information were employed to search for possible subrecent small-scale vertical movement along the anticlinal fold belt of the central Negev, Israel. Such tectonic deformation might indicate displacement on the buried reverse faults underneath the anticlines. Variations in the thickness of the alluvial fill in the study area, which are in accordance with the fold structures, could be an indication of recent folding activity along the anticlinal system. In order to detect these thickness variations in the alluvial fill, seismic refraction and electrical resistivity measurements were carries out along the valley of Nahal Besor, which crosses the anticlinal belt. The thickness variations of the alluvial fill along the valley were not found to indicate any significant tectonic movement along the anticlines during the Pleistocene. The thickest alluvium was found overlying a karst bedrock, hence karst relief is suggested to be responsible for these variations.

  1. 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.

  2. 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.

  3. Active and legacy mining in an arid urban environment: challenges and perspectives for Copiapó, Northern Chile.

    PubMed

    Carkovic, Athena B; Calcagni, Magdalena S; Vega, Alejandra S; Coquery, Marina; Moya, Pablo M; Bonilla, Carlos A; Pastén, Pablo A

    2016-08-01

    Urban expansion in areas of active and legacy mining imposes a sustainability challenge, especially in arid environments where cities compete for resources with agriculture and industry. The city of Copiapó, with 150,000 inhabitants in the Atacama Desert, reflects this challenge. More than 30 abandoned tailings from legacy mining are scattered throughout its urban and peri-urban area, which include an active copper smelter. Despite the public concern generated by the mining-related pollution, no geochemical information is currently available for Copiapó, particularly for metal concentration in environmental solid phases. A geochemical screening of soils (n = 42), street dusts (n = 71) and tailings (n = 68) was conducted in November 2014 and April 2015. Organic matter, pH and elemental composition measurements were taken. Notably, copper in soils (60-2120 mg/kg) and street dusts (110-10,200 mg/kg) consistently exceeded international guidelines for residential and industrial use, while a lower proportion of samples exceeded international guidelines for arsenic, zinc and lead. Metal enrichment occurred in residential, industrial and agricultural areas near tailings and the copper smelter. This first screening of metal contamination sets the basis for future risk assessments toward defining knowledge-based policies and urban planning. Challenges include developing: (1) adequate intervention guideline values; (2) appropriate geochemical background levels for key metals; (3) urban planning that considers contaminated areas; (4) cost-effective control strategies for abandoned tailings in water-scarce areas; and (5) scenarios and technologies for tailings reprocessing. Assessing urban geochemical risks is a critical endeavor for areas where extreme events triggered by climate change are likely, as the mud flooding that impacted Copiapó in late March 2015.

  4. Superoxide anion radical scavenging activities of herbs and pastures in northern Japan determined using electron spin resonance spectrometry.

    PubMed

    Al-Mamun, Mohammad; Yamaki, Koji; Masumizu, Toshiki; Nakai, Yumi; Saito, Katsumi; Sano, Hiroaki; Tamura, Yoshifumi

    2007-07-30

    Free radicals are not only destructive to the living cells but also reduce the quality of animal products through oxidation. As a result the superoxide anion radical (O2-), one of the most destructive reactive oxygen species, is a matter of concern for the animal scientists as well as feed manufacturers to ensure the quality of product to reach consumers demand. The superoxide anion radical scavenging activities (SOSA) of water and MeOH extracts of 2 herbs and 9 pasture samples collected from lowland and highland swards were determined against a 5,5-dimethyl-1-pyroline-N-oxide-O2-spin adduct based on a hypoxanthine-xanthine oxidase reaction using electron spin resonance spectrometry. Both the water and MeOH extracted SOSA differed among the herbs and pastures. Species and altitudinal variations were observed between extraction methods. The herbs were higher in both water and MeOH extracted SOSA than the pastures except for water extracts of one pasture, white clover (Trifolium repens L.). Among the pastures, quackgrass (Agrophyron repens L.) showed higher SOSA in both the MeOH and water extracts, and timothy (Phleum pretense L.) showed higher MeOH extracted SOSA. It is apparent that the kind and amount of antioxidants differ among herbs and pastures. Animal health and quality of animal products could be improved by adequate selection and combining of herbs and pastures having higher SOSA.

  5. Long-lasting Cadomian magmatic activity along an active northern Gondwana margin: U-Pb zircon and Sr-Nd isotopic evidence from the Brunovistulian Domain, eastern Bohemian Massif

    NASA Astrophysics Data System (ADS)

    Soejono, Igor; Janoušek, Vojtěch; Žáčková, Eliška; Sláma, Jiří; Konopásek, Jiří; Machek, Matěj; Hanžl, Pavel

    2016-11-01

    Cadomian magmatic complexes of the Brunovistulian Domain crop out at the eastern termination of the Bohemian Massif. However, the age, nature and geotectonic affinity of some of pre-Variscan (meta-)igneous rock complexes from this domain are still unknown. Geochronological and geochemical study of the granitic rocks across the Brunovistulian Domain reveals new information about the timing and nature of this magmatic activity originally situated along the northern margin of Gondwana. Zircon U-Pb data (601 ± 3 Ma, Brno Massif; 634 ± 6 Ma, paraautochtonous core of the Svratka Dome; 568 ± 3 Ma, Bíteš orthogneiss) from the allochtonous Moravicum indicate the prolonged magmatic activity within the Brunovistulian Domain during the Ediacaran. The major- and trace-element and Sr-Nd isotopic signatures show heterogeneous geochemical characteristics of the granitic rocks and suggest a magmatic-arc geotectonic setting. The two-stage Depleted Mantle Nd model ages (c. 1.3-2.0 Ga) indicate derivation of the granitic rocks from a relatively primitive crustal source, as well as from an ancient and evolved continental crust of the Brunovistulian Domain. These results constrain the magmatic-arc activity to c. 635-570 Ma and provide a further evidence for a long-lived (at least c. 65 Myr) and likely episodic subduction-related magmatism at the northern margin of Gondwana. The presence of granitic intrusions derived from variously mature crustal sources at different times suggests heterogeneous crustal segments to having been involved in the magmatic-arc system during its multistage evolution.

  6. New insights into fault activation and stress transfer between en echelon thrusts: The 2012 Emilia, Northern Italy, earthquake sequence

    NASA Astrophysics Data System (ADS)

    Cheloni, D.; Giuliani, R.; D'Agostino, N.; Mattone, M.; Bonano, M.; Fornaro, G.; Lanari, R.; Reale, D.; Atzori, S.

    2016-06-01

    Here we present the results of the inversion of a new geodetic data set covering the 2012 Emilia seismic sequence and the following 1 year of postseismic deformation. Modeling of the geodetic data together with the use of a catalog of 3-D relocated aftershocks allows us to constrain the rupture geometries and the coseismic and postseismic slip distributions for the two main events (Mw 6.1 and 6.0) of the sequence and to explore how these thrust events have interacted with each other. Dislocation modeling reveals that the first event ruptured a slip patch located in the center of the Middle Ferrara thrust with up to 1 m of reverse slip. The modeling of the second event, located about 15 km to the southwest, indicates a main patch with up to 60 cm of slip initiated in the deeper and flatter portion of the Mirandola thrust and progressively propagated postseismically toward the top section of the rupture plane, where most of the aftershocks and afterslip occurred. Our results also indicate that between the two main events, a third thrust segment was activated releasing a pulse of aseismic slip equivalent to a Mw 5.8 event. Coulomb stress changes suggest that the aseismic event was likely triggered by the preceding main shock and that the aseismic slip event probably brought the second fault closer to failure. Our findings show significant correlations between static stress changes and seismicity and suggest that stress interaction between earthquakes plays a significant role among continental en echelon thrusts.

  7. 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.

  8. 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.

  9. 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.

  10. 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

  11. 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).

  12. 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.

  13. 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.

  14. 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

  15. 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

  16. 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.

  17. 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.

  18. 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

  19. 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.

  20. 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.

  1. Discovery of an active shallow submarine silicic volcano in the northern Izu-Bonin Arc: volcanic structure and potential hazards of Oomurodashi Volcano (Invited)

    NASA Astrophysics Data System (ADS)

    Tani, K.; Ishizuka, O.; Nichols, A. R.; Hirahara, Y.; Carey, R.; McIntosh, I. M.; Masaki, Y.; Kondo, R.; Miyairi, Y.

    2013-12-01

    Oomurodashi is a bathymetric high located ~20 km south of Izu-Oshima, an active volcanic island of the northern Izu-Bonin Arc. Using the 200 m bathymetric contour to define its summit dimensions, the diameter of Oomurodashi is ~20 km. Oomurodashi has been regarded as inactive, largely because it has a vast flat-topped summit at 100 - 150 meters below sea level (mbsl). During cruise NT07-15 of R/V Natsushima in 2007, we conducted a dive survey in a small crater, Oomuro Hole, located in the center of the flat-topped summit, using the remotely-operated vehicle (ROV) Hyper-Dolphin. The only heat flow measurement conducted on the floor of Oomuro Hole during the dive recorded an extremely high value of 4,200 mW/m2. Furthermore, ROV observations revealed that the southwestern wall of Oomuro Hole consists of fresh rhyolitic lavas. These findings suggest that Oomurodashi is in fact an active silicic submarine volcano. To confirm this hypothesis, we conducted detailed geological and geophysical ROV Hyper-Dolphin (cruise NT12-19). In addition to further ROV surveys, we carried out single-channel seismic (SCS) surveys across Oomurodashi in order to examine the shallow structures beneath the current edifice. The ROV surveys revealed numerous active hydrothermal vents on the floor of Oomuro Hole, at ~200 mbsl, with maximum water temperature measured at the hydrothermal vents reaching 194°C. We also conducted a much more detailed set of heat flow measurements across the floor of Oomuro Hole, detecting very high heat flows of up to 29,000 mW/m2. ROV observations revealed that the area surrounding Oomuro Hole on the flat-topped summit of Oomurodashi is covered by extensive fresh rhyolitic lava and pumice clasts with minimum biogenetic or manganese cover, suggesting recent eruption(s). These findings strongly indicate that Oomurodashi is an active silicic submarine volcano, with recent eruption(s) occurring from Oomuro Hole. Since the summit of Oomurodashi is in shallow water, it

  2. A Comparison of Structural Data and Seismic Images For Low-Angle Normal Faults in the Northern Apennines (Central Italy): Constraints on Geometry and Activity

    NASA Astrophysics Data System (ADS)

    Collettini, C.; Barchi, M. R.

    2001-12-01

    During the last 20 Myr extensional tectonics in the Northern Apennines have moved from the Tyrrhenian sea toward east. Much of the extension is due to low-angle east-dipping normal faults now exhumed in the Tyrrhenian islands and Tuscany, while still accommodating deformation in the Apenninic chain (Umbria region 200 km eastward). This tectonic framework provide an example where exhumed structures can be compared with active extensional structures and processes affecting the Umbria region. It is here proposed the case study of two of these low angle normal faults, the Zuccale fault (Zf), cropping out in the Elba island and the Altotiberina fault (ATF) mainly detected by seismic profiles crossing the Umbria region. The Zf in the eastern part of the Elba island juxtaposes along a gently ( ~ 10° ) eastward dipping contact, the Upper Cretaceous Helminthoid flysch in its hangingwall over the Permian-Triassic (?) phyllitic basement in its footwall. Structural analysis of the brittle structures that characterise the fault zone has been used to constraint the state of stress under which the fault slipped. From the N-S trending vertical vein system perpendicular to the slickenlines of the fault plane and from the Andersonian normal faults present within the fault gouge, some of them rotated according to a top to the east movement, we infer that (1) the maximum principal stress was sub vertical during the fault activity (2) the fault accommodate slip under low values of differential stress and at dips similar to its present flat geometry (3) local fluid overpressures were attained during the fault activity favoured by a thick fault gouge. The geological scenario described in the Elba island shows similarities with the active deformation of the Umbria region. Seismic profiles crossing this area matched with surface geology highlight the presence of an east-dipping low-angle ( ~ 20° ) normal fault, the Altotiberina fault (ATF), and antithetic seismogenic structures bounding

  3. Microbial Activity Response to Solar Radiation across Contrasting Environmental Conditions in Salar de Huasco, Northern Chilean Altiplano

    PubMed Central

    Hernández, Klaudia L.; Yannicelli, Beatriz; Olsen, Lasse M.; Dorador, Cristina; Menschel, Eduardo J.; Molina, Verónica; Remonsellez, Francisco; Hengst, Martha B.; Jeffrey, Wade H.

    2016-01-01

    In high altitude environments, extreme levels of solar radiation and important differences of ionic concentrations over narrow spatial scales may modulate microbial activity. In Salar de Huasco, a high-altitude wetland in the Andean mountains, the high diversity of microbial communities has been characterized and associated with strong environmental variability. Communities that differed in light history and environmental conditions, such as nutrient concentrations and salinity from different spatial locations, were assessed for bacterial secondary production (BSP, 3H-leucine incorporation) response from short-term exposures to solar radiation. We sampled during austral spring seven stations categorized as: (a) source stations, with recently emerged groundwater (no-previous solar exposure); (b) stream running water stations; (c) stations connected to source waters but far downstream from source points; and (d) isolated ponds disconnected from ground sources or streams with a longer isolation and solar exposure history. Very high values of 0.25 μE m-2 s-1, 72 W m-2 and 12 W m-2 were measured for PAR, UVA, and UVB incident solar radiation, respectively. The environmental factors measured formed two groups of stations reflected by principal component analyses (near to groundwater sources and isolated systems) where isolated ponds had the highest BSP and microbial abundance (35 microalgae taxa, picoeukaryotes, nanoflagellates, and bacteria) plus higher salinities and PO43- concentrations. BSP short-term response (4 h) to solar radiation was measured by 3H-leucine incorporation under four different solar conditions: full sun, no UVB, PAR, and dark. Microbial communities established in waters with the longest surface exposure (e.g., isolated ponds) had the lowest BSP response to solar radiation treatments, and thus were likely best adapted to solar radiation exposure contrary to ground source waters. These results support our light history (solar exposure) hypothesis

  4. Microbial Activity Response to Solar Radiation across Contrasting Environmental Conditions in Salar de Huasco, Northern Chilean Altiplano.

    PubMed

    Hernández, Klaudia L; Yannicelli, Beatriz; Olsen, Lasse M; Dorador, Cristina; Menschel, Eduardo J; Molina, Verónica; Remonsellez, Francisco; Hengst, Martha B; Jeffrey, Wade H

    2016-01-01

    In high altitude environments, extreme levels of solar radiation and important differences of ionic concentrations over narrow spatial scales may modulate microbial activity. In Salar de Huasco, a high-altitude wetland in the Andean mountains, the high diversity of microbial communities has been characterized and associated with strong environmental variability. Communities that differed in light history and environmental conditions, such as nutrient concentrations and salinity from different spatial locations, were assessed for bacterial secondary production (BSP, (3)H-leucine incorporation) response from short-term exposures to solar radiation. We sampled during austral spring seven stations categorized as: (a) source stations, with recently emerged groundwater (no-previous solar exposure); (b) stream running water stations; (c) stations connected to source waters but far downstream from source points; and (d) isolated ponds disconnected from ground sources or streams with a longer isolation and solar exposure history. Very high values of 0.25 μE m(-2) s(-1), 72 W m(-2) and 12 W m(-2) were measured for PAR, UVA, and UVB incident solar radiation, respectively. The environmental factors measured formed two groups of stations reflected by principal component analyses (near to groundwater sources and isolated systems) where isolated ponds had the highest BSP and microbial abundance (35 microalgae taxa, picoeukaryotes, nanoflagellates, and bacteria) plus higher salinities and PO4(3-) concentrations. BSP short-term response (4 h) to solar radiation was measured by (3)H-leucine incorporation under four different solar conditions: full sun, no UVB, PAR, and dark. Microbial communities established in waters with the longest surface exposure (e.g., isolated ponds) had the lowest BSP response to solar radiation treatments, and thus were likely best adapted to solar radiation exposure contrary to ground source waters. These results support our light history (solar exposure

  5. Provenance of a large Lower Cretaceous turbidite submarine fan complex on the active Laurasian margin: Central Pontides, northern Turkey

    NASA Astrophysics Data System (ADS)

    Akdoğan, Remziye; Okay, Aral I.; Sunal, Gürsel; Tari, Gabor; Meinhold, Guido; Kylander-Clark, Andrew R. C.

    2017-02-01

    The Pontides formed the southern active margin of Laurasia during the Mesozoic. They became separated from mainland Laurasia during the Late Cretaceous, with the opening of the Black Sea as an oceanic back-arc basin. During the Early Cretaceous, a large submarine turbidite fan complex developed in the Central Pontides. The turbidites cover an area of 400 km by 90 km with a thickness of more than 2 km. We have investigated the provenance of these turbidites-the Çağlayan Formation-using paleocurrent measurements, U-Pb detrital zircon ages, REE abundances of dated zircons and geochemistry of detrital rutile grains. 1924 paleocurrent measurements from 96 outcrop stations indicate flow direction from northwest to southeast in the eastern part of the Çağlayan Basin and from north-northeast to west-southwest in the western part. 1194 detrital zircon ages from 13 Lower Cretaceous sandstone samples show different patterns in the eastern, central and western parts of the basin. The majority of the U-Pb detrital zircon ages in the eastern part of the basin are Archean and Paleoproterozoic (61% of all zircon ages, 337 grains); rocks of these ages are absent in the Pontides and present in the Ukrainian Shield, which indicates a source north of the Black Sea. In the western part of the basin the majority of the zircons are Carboniferous and Neoproterozoic (68%, 246 grains) implying more local sources within the Pontides. The detrital zircons from the central part show an age spectrum as mixture of zircons from western and eastern parts. Significantly, Jurassic and Early Cretaceous zircons make up less than 2% of the total zircon population, which implies lack of a coeval magmatic arc in the region. This is compatible with the absence of the Lower Cretaceous granites in the Pontides. Thus, although the Çağlayan Basin occupied a fore-arc position above the subduction zone, the arc was missing, probably due to flat subduction, and the basin was largely fed from the Ukrainian

  6. Permafrost vulnerability and active layer thickness increases over the high northern latitudes inferred from satellite remote sensing and process model assessments

    NASA Astrophysics Data System (ADS)

    Park, Hotaek; Kim, Youngwook

    2016-04-01

    Permafrost extent (PE) and active layer thickness (ALT) are important for assessing high northern latitude (HNL) ecological and hydrological processes, and potential land-atmosphere carbon and climate feedbacks. We developed a new approach to infer PE from satellite microwave remote sensing of daily landscape freeze-thaw (FT) status. Our results document, for the first time, the use of satellite microwave FT observations for monitoring permafrost extent and condition. The FT observations define near-surface thermal status used to determine permafrost extent and stability over a 30-year (1980-2009) satellite record. The PE results showed similar performance against independent inventory and process model (CHANGE) estimates, but with larger differences over heterogeneous permafrost subzones. A consistent decline in the ensemble mean of permafrost areas (-0.33 million km2 decade-1; p < 0.05) coincides with regional warming (0.4 °C decade-1; p < 0.01), while more than 40% (9.6 million km2) of permafrost areas are vulnerable to degradation based on the 30-year PE record. ALT estimates determined from satellite (MODIS) and ERA-Interim temperatures, and CHANGE simulations, compared favorably with independent field observations and indicate deepening ALT trends consistent with widespread permafrost degradation under recent climate change. The integration of remote sensing and modeling of permafrost and active layer conditions developed from this study may facilitate regular and effective regional monitoring of these parameters, and expand applications of remote sensing for examining permafrost-related feedbacks and consequences for biogeochemical and hydrological cycling in the Arctic.

  7. Improving diet and activity of insectivorous primates in captivity: Naturalizing the diet of Northern Ceylon gray slender loris, Loris lydekkerianus nordicus.

    PubMed

    Williams, Emma; Cabana, Francis; Nekaris, K A I

    2015-01-01

    Data on in-situ diet and nutritional requirements should inform the provision of food to captive insectivorous primates. Despite the growing availability of such information an over-reliance on commercially available primate foods and fruit continues in many captive establishments. Wild slender lorises are almost exclusively insectivorous, yet captive conspecifics are fed a primarily frugivorous diet that is likely to contribute to behavioral and health problems. We investigated the effect of naturalizing diet in the Northern Ceylon grey slender loris (Loris lydekkerianus nordicus) by providing live insect prey to a captive group of five individuals. We calculated activity budgets in accordance with six established categories and recorded positional behaviors. We collected data over 30 hours for each of three conditions: pre-enrichment, enrichment, post-enrichment. We hypothesized that increased opportunity for the display of natural behaviors would be stimulated by the dietary enrichment of live insects and made the following predictions; 1) Percentage time spent foraging would increase and time spent inactive would decrease; 2) behavioral repertoires would increase; 3) foraging patterns would be more constant over time with reduced feeding-time peaks. We analyzed time budget and behavioral changes using Friedman tests. We found significant changes in activity budgets with inactivity reduced and foraging levels increased to levels seen in wild slender lorises. We found a significant increase in postures used in foraging and a wider behavioral repertoire. We discuss the benefits of providing free-ranging live food in relation to enhancing the temporal-spatial distribution of food acquisition, satisfying nutritional requirements, balancing energy intake, and expenditure, expanding sensory stimulation, and promoting behavioral competence. We discuss our findings in relation to other insectivorous primates.

  8. Chemical correlation of some late Cenozoic tuffs of Northern and Central California by neutron activation analysis of glass and comparison with X-ray fluorescence analysis

    USGS Publications Warehouse

    Sarna-Wojcicki, Andrei M.; Bowman, Harry W.; Russell, Paul C.

    1979-01-01

    Glasses separated from several dacitic and rhyolitic late Cenozoic tuffs of northern and central California were analyzed by neutron activation for more than 43 elemental abundances. Eighteen elements--scandiurn, manganese, iron, zinc, rubidium, cesium, barium, lanthanum, cerium, samarium, europium, terbiurn, dysprosiurn, ytterbiurn, hafniurn, tantalurn, thorium and uranium--were selected as most suitable for purposes of chemical correlation on the basis of their natural variability in silicic tuffs and the precision obtainable in analysis. Stratigraphic relations between tuffs and replicate chemical analyses on individual tuffs make it possib1e to calibrate a quantitative parameter, the similarity coefficient, which indicates the degree of correlation for the tuffs studied. The highest similarity coefficient (0.99) was obtained for analyses of two tuffs (potassium-argon dated at about' 6.0 m.y.) exposed in the Merced(?) and Petaluma Formations of Sonoma County, which represent different paleoenvironments, shallow-water marine and fresh water or brackish marine, respectively. Corre1ation of these formations on the basis of criteria other than tephrochronoloqy would be difficult. Results of neutron activation analysis in general confirm earlier correlations made on the basis of analysis by X-ray fluorescence but also make it possible to resolve small compositional differences between chemically simi1ar tuffs in stratigraphic proximity. The Lawlor Tuff (potassium-argon dated at about 4.0 m.y.) is identified at two new localities: in a core sample obtained from a bore hole east of Suisun Bay, and from the Kettleman Hills of western San Joaquin Valley. This identification permits correlation of the uppermost part of the marine Etchegoin Formation in the San Joaquin Valley with the continental Livermore Gravels of Clark, the Tassajara Formation, and the upper part of the Sonoma Volcanics in the cel1tral Coast Ranges of California. A younger tuff near the top of the

  9. Parameters of induced and natural seismicity recorded in the vicinity of an active low angle normal fault in the Northern Apennines (Italy)

    NASA Astrophysics Data System (ADS)

    Braun, T.; Cesca, S.; Martirosian, A.; Dahm, T.

    2012-12-01

    The Upper Tiber Valley is situated in the northern part of the Central Apennines and is setting of a number of geological phenomena, like CO2 degassing, moderate earthquakes (M < 6) and a strong microseismicity. The major part of the recorded seismicity can be associated to a Low Angle Normal Fault (LANF) - the so called Alto Tiberina Fault - but some of the recorded seismograms show signals similar to those recorded on active volcanoes. In the vicinity of the ATF human activity provides a number of candidates, capable to influence the local stress regime and the seismic release in the area: (i) a huge barrier lake is directly situated on the ATF and is characterized by significant seasonal water level oscillations, (ii) a cement plant and decommissioned mines present in the area are in the direct vicinity of epicentres of tornillo-like seismograms and episodes of non-volcanic tremor, (iii) since July 2011, a private company extracts CO2 by a 4 km deep borehole (PSS1). Since 2003 different seismic network and array configurations have been deployed to monitor the local seismicity. Human related influences, as realized by the industrial activities of cement plants, quarries or superficial mines may produce seismic signals, but will not directly have an impact on the mechanical behaviour of an active fault system at crustal depth. However, water level changes in the huge barrier lake or long term CO2-extraction from the upper crust have the capability to directly influence the stress field at depth. Since summer 2011 the reservoir user produces about 4 tons per hour of reservoir gases for commercial usage and trading. This production lead to a pore pressure change and slow depletion of the reservoir formation, similar to many other gas fields under production. Since production volumes and pore pressure changes are relatively well known, we consider the local depletion-induced stress changes on the ATF and in the surrounding rock as driving forces to the system. The

  10. Exposure of northern leopard frogs in the Green Bay ecosystem to polychlorinated biphenyls, polychlorinated dibenzo-p-dioxins, and polychlorinated dibenzofurans is measured by direct chemistry but not hepatic ethoxyresorufin-O-deethylase activity

    SciTech Connect

    Huang, Y.W.; Karasov, W.H.; Patnode, K.A.; Jefcoate, C.R.

    1999-10-01

    The authors measured concentrations of polychlorinated biphenyls (PCBs), polychlorinated dibenzo-p-dioxins (PCDDs), and polychlorinated dibenzofurans (PCDFs) in northern leopard frogs collected from the Green Bay ecosystem and explored the catalytic activity of hepatic cytochrome P450-associated monooxygenase (P450 enzyme) as a biomarker for exposure to aryl hydrocarbon receptor (AhR) agonists. The two hypotheses tested were PCH concentrations in northern leopard frogs would be positively correlated with sediment polychlorinated hydrocarbon (PCH) levels in wetland habitats along a contamination gradient and hepatic ethoxyresorufin-O-deethylase (EROD) activity of northern leopard frogs, which is presumably mediated by aryl hydrocarbon receptor (AhR), would be positively correlated with PCH concentrations in frog carcasses from different collection sites. In 1994 and 1995, frogs from seven sites along the lower Fox River and Green Bay, USA, were assayed for hepatic EROD activities and whole carcass concentrations of PCBs, PCDDs, and PCDFs. Tissue total PCB concentrations ranging from 3 to 154 ng/g were significantly correlated with sediment PCB levels. Only one PCDD and two PCDFs at concentrations of 6 to 8 pg/g were found in the frogs collected with frog body weight and was similar among sites except for Peter's Marsh. No significant correlation was found between EROD activity and carcass PCB concentration. This result was consistent with the fact that the frogs collected from the Green Bay ecosystem had relatively low PCB concentrations compared with what was required for induction in the laboratory.

  11. 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

  12. 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

  13. 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.

  14. 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.

  15. 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.

  16. 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

  17. 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.

  18. 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

  19. Interannual variation of rare earth element abundances in corals from northern coast of the South China Sea and its relation with sea-level change and human activities

    USGS Publications Warehouse

    Liu, Yajing; Peng, Z.; Wei, G.; Chen, T.; Sun, W.; He, J.; Liu, Gaisheng; Chou, C.-L.; Shen, C.-C.

    2011-01-01

    Here we present interannual rare earth element (REE) records spanning the last two decades of the 20th century in two living Porites corals, collected from Longwan Bay, close to the estuarine zones off Wanquan River of Hainan Island and Hong Kong off the Pearl River Delta of Guangdong Province in the northern South China Sea. The results show that both coral REE contents (0.5-40 ng g-1 in Longwan Bay and 2-250 ng g-1 in Hong Kong for La-Lu) are characterized with a declining trend, which are significantly negative correlated with regional sea-level rise (9.4 mm a-1 from 1981 to 1996 in Longwan Bay, 13.7 mm a-1 from 1991 to 2001 in Hong Kong). The REE features are proposed to be resulted from seawater intrusion into the estuaries in response to contemporary sea-level rise. However, the tendency for the coral Er/Nd time series at Hong Kong site is absent and there is no significant relation between Er/Nd and total REEs as found for the coral at Longwan Bay site. The observations are likely attributed to changes of the water discharge and sediment load of Pearl River, which have been significantly affected by intense human activities, such as the construction of dams/reservoirs and riverbed sediment mining, in past decades. The riverine sediment load/discharge ratio of the Pearl River decreased sharply with a rate of 0.02 kg m-3 a-1, which could make significant contribution to the declining trend of coral REE. We propose that coastal corals in Longwan Bay and similar unexplored sites with little influences of river discharge and anthropogenic disruption are ideal candidates to investigate the influence of sea-level change on seawater/coral REE. ?? 2010 Elsevier Ltd.

  20. History of human activity in last 800 years reconstructed from combined archive data and high-resolution analyses of varved lake sediments from Lake Czechowskie, Northern Poland

    NASA Astrophysics Data System (ADS)

    Słowiński, Michał; Tyszkowski, Sebastian; Ott, Florian; Obremska, Milena; Kaczmarek, Halina; Theuerkauf, Martin; Wulf, Sabine; Brauer, Achim

    2016-04-01

    The aim of the study was to reconstruct human and landscape development in the Tuchola Pinewoods (Northern Poland) during the last 800 years. We apply an approach that combines historic maps and documents with pollen data. Pollen data were obtained from varved lake sediments at a resolution of 5 years. The chronology of the sediment record is based on varve counting, AMS 14C dating, 137Cs activity concentration measurements and tephrochronology (Askja AD 1875). We applied the REVEALS model to translate pollen percentage data into regional plant abundances. The interpretation of the pollen record is furthermore based on pollen accumulation rate data. The pollen record and historic documents show similar trends in vegetation development. During the first phase (AD 1200-1412), the Lake Czechowskie area was still largely forested with Quercus, Carpinus and Pinus forests. Vegetation was more open during the second phase (AD 1412-1776), and reached maximum openness during the third phase (AD 1776-1905). Furthermore, intensified forest management led to a transformation from mixed to pine dominated forests during this period. Since the early 20th century, the forest cover increased again with dominance of the Scots pine in the stand. While pollen and historic data show similar trends, they differ substantially in the degree of openness during the four phases with pollen data commonly suggesting more open conditions. We discuss potential causes for this discrepancy, which include unsuitable parameters settings in REVEALS and unknown changes in forest structure. Using pollen accumulation data as a third proxy record we aim to identify the most probable causes. Finally, we discuss the observed vegetation change in relation the socio-economic development of the area. This study is a contribution to the Virtual Institute of Integrated Climate and Landscape Evolution Analysis - ICLEA- of the Helmholtz Association and National Science Centre, Poland (grant No. 2011/01/B/ST10

  1. Interannual variation of rare earth element abundances in corals from northern coast of the South China Sea and its relation with sea-level change and human activities.

    PubMed

    Liu, Yi; Peng, Zicheng; Wei, Gangjian; Chen, Tegu; Sun, Weidong; He, Jianfeng; Liu, Guijian; Chou, Chen-Lin; Shen, Chuan-Chou

    2011-02-01

    Here we present interannual rare earth element (REE) records spanning the last two decades of the 20th century in two living Porites corals, collected from Longwan Bay, close to the estuarine zones off Wanquan River of Hainan Island and Hong Kong off the Pearl River Delta of Guangdong Province in the northern South China Sea. The results show that both coral REE contents (0.5-40 ng g⁻¹ in Longwan Bay and 2-250 ng g⁻¹ in Hong Kong for La-Lu) are characterized with a declining trend, which are significantly negative correlated with regional sea-level rise (9.4 mm a⁻¹) from 1981 to 1996 in Longwan Bay, 13.7 mm a⁻¹ from 1991 to 2001 in Hong Kong). The REE features are proposed to be resulted from seawater intrusion into the estuaries in response to contemporary sea-level rise. However, the tendency for the coral Er/Nd time series at Hong Kong site is absent and there is no significant relation between Er/Nd and total REEs as found for the coral at Longwan Bay site. The observations are likely attributed to changes of the water discharge and sediment load of Pearl River, which have been significantly affected by intense human activities, such as the construction of dams/reservoirs and riverbed sediment mining, in past decades. The riverine sediment load/discharge ratio of the Pearl River decreased sharply with a rate of 0.02 kg m⁻³ a⁻¹, which could make significant contribution to the declining trend of coral REE. We propose that coastal corals in Longwan Bay and similar unexplored sites with little influences of river discharge and anthropogenic disruption are ideal candidates to investigate the influence of sea-level change on seawater/coral REE.

  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. 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.

  6. Developing a Small-Scale De-Fluoridation Filter for Use in Rural Northern Ghana with Activated Alumina As the Sorbent

    NASA Astrophysics Data System (ADS)

    Craig, L.; Stillings, L. L.

    2014-12-01

    In northern Ghana, groundwater is the main source of household water and is generally considered safe to drink. However in some areas it contains fluoride (F-) concentrations above the 1.5 ppm limit recommended by the World Health Organization, putting the users at risk of fluorosis. The study area in the Upper East Region of Ghana has pockets of groundwater F- up to 4.6 ppm and, as a result, also has a high percentage of residents with dental fluorosis. They have no alternative water source and, because of poverty and limited access to technology, lack the capacity to set up advanced treatment systems. One proposed solution is to attach F- adsorption filters to the wells, since adsorption is considered a simple and cost effective approach for treating high F-drinking water. This study evaluates activated alumina as a sorbent for use in de-fluoridation filters in the study area. We evaluated the long-term adsorption capacity of activated alumina, and changes in F- adsorption rate and capacity with grain size. We measured differences in positive surface charge (C m-2) via slow acid titration, as well as F- loading with varied prior hydration time. Results from this research show no notable change in F- adsorption or positive surface charge when the activated alumina surface was pre-equilibrated in distilled water from 24 hours to 30 weeks. The results of F- loading show a maximum of ~3.4 mg F- sorbed per gm activated alumina (initial pH ~6.9, initial F- 1 to 60 ppm, 20 hr reaction time). The pH dependent surface charge is ~0.14 C m-2 at pH of ~4.4 and is zero at pH ~8.6. F- loading experiments were conducted with grain size 0.125 to 0.250 mm and 0.5 to 1.0 mm to evaluate changes in F- adsorption rate (initial pH ~6.9, initial F- 10 ppm) and F- loading (initial pH ~6.9, initial F- 1 to 60 ppm, 20 hr reaction time). The F- loading did not change with grain size. However time to equilibrium increased dramatically with a decrease in grain size - after one hour of

  7. 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.

  8. 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.

  9. Coseismic uplift and fault model of marine active faults in 1729 AD revealed by fossilized intertidal sessile organisms along the northern coast of the Noto Peninsula, central Japan

    NASA Astrophysics Data System (ADS)

    Hamada, M.; Hiramatsu, Y.; Oda, M.; Yamaguchi, H.

    2015-12-01

    The Noto Peninsula is located in the backarc region of southwest Japan and is characterized by geomorphologic features formed by active tectonics and glacial eustasy through the Quaternary. Pleistocene marine terraces along the northern coast of the Noto Peninsula indicate uplift in the coastal area through the late Quaternary (Ota and Hirakawa, 1979). Recently, an active fault zone on the seafloor off the coast was found and was divided into four segments, Monzen-oki, Saruyama-oki, Wajima-oki, and Suzu-oki, from west to east (Inoue and Okamura, 2010). We investigated vertical displacement along the coast using intertidal sessile organisms at nine sites on the rocky coast. We measured the height of fossilized Pomatoleios kraussii by GPS surveying together with a sea-level change curve, and dated them using the AMS 14C method. The vertical displacements and dates at the sites implied that coastal uplift occurred along 20 km of coastline, corresponding to the Wajima-oki segment zone, and most likely between 1600 and 1800 AD. This is coincident with seismic damage in this area in 1729 AD recorded in historical documents. We constructed a fault model with three rectangular faults in a homogeneous elastic half-space and estimated the optimal net slip and rake by a non-linear inversion method (Matsu'ura and Hasegawa, 1987). The best fit to the estimated vertical displacements is provided by a net slip of 1.8 m with a rake of 90° for the western fault plane and a net slip of 0.6 m with a rake of 90° for the center and the eastern fault planes. The moment magnitude (Mw) calculated from these parameters with a rigidity of 30 GPa is 6.6. We compared the elevation distribution of the former shorelines based on coastal terraces and the 1729 earthquake uplifts. Assuming that the coastal uplift is caused by the cumulative crustal deformation produced by the same size event as the 1729 earthquake, the average recurrence interval of the events is estimated to be 1700 years.

  10. Large-scale thrusting along the northern margin of the Tibetan Plateau and the southwest Tarim basin: 230 km long active Hotian thrust sheet

    NASA Astrophysics Data System (ADS)

    Suppe, J.; Wang, X.; He, D.; Liang, H.

    2015-12-01

    We present the geometry, kinematics and mechanics of large-scale active thrusting in the western Kunlunshan and southwest Tarim basin, which accounts for ~130-165km total shortening of Tarim crust at the northern margin of Tibet. The great frontal structure is the ~230km long bedding-parallel Hotian thrust sheet, which is perhaps the longest active intact thrust sheet in the world, composed of flat-lying strata of the Tarim basin sliding northward on a regional gypsum detachment at the base of the Cenozoic sequence. The toe of the Hotian thrust ramps to the surface two thirds of the way across the Tarim basin, forming the Selibuya-Mazartag hills in the Taklamakan sand desert. At the southern edge of the Tarim basin in the Kunlunshan foothills, a set of high-amplitude anticlines are growing by complex break-forward ramping and wedging in the Hotian thrust sheet as it steps up to the Cenozoic gypsum detachment from a regional Cambrian evaporate detachment that extends under Tibet. More interior structures such as the Tiklik thrust bring older strata and Proterozoic basement to the surface, together with their Cenozoic Tarim cover in the Buya basin. The Cambrian detachment also extends northward under the Tarim basin with minor hanging-wall deformation that locally warps the overlying Hotian thrust sheet, producing a complete syntectonic record in seismically imaged growth strata of its northward motion over these warps. Seismic profiles in the southwest Tarim foothill belt also reveal widespread growth strata that record much of the structural history beginning in the early Pliocene Atushi Formation. Ages of seismic reflectors are calibrated to a surface magnetostratigraphic sequence (Zheng et al., 2000). The beginning of thrusting and folding in the southwest Tarim basin north of the Tiklik thrust is dated at 3.6Ma with shortening >25km and a progressive northward propagation toward the Selibuya-Mazartag hills. The overall shortening rate is ~10 mm/yr. The gypsum

  11. Faults paragenesis and paleostress state in the zone of actively propagating continental strike-slip on the example of North Khangai fault (Northern Mongolia)

    NASA Astrophysics Data System (ADS)

    Sankov, Vladimir; Parfeevets, Anna

    2014-05-01

    Sublatitudinal North Khangai fault extends from Ubsunuur basin to the eastern part of the Selenga corridor trough 800 km. It is the northern boundary of the massive Mongolian block and limits of the Baikal rift system structures propagation in the south (Logatchev, 2003). Late Cenozoic and present-day fault activity are expressed in the left-lateral displacements of a different order of river valleys and high seismicity. We have carried out studies of the kinematics of active faults and palaeostresses reconstruction in the zone of the dynamic influence of North Khangai fault, the width of which varies along the strike and can exceeds 100 km. The result shows that the fault zone has a longitudinal and a transverse zoning. Longitudinal zonation presented gradual change from west to east regions of compression and transpression regimes (Khan-Khukhey ridge) to strike-slip regime (Bolnay ridge) and strike-slip and transtensive regimes (west of Selenga corridor). Strike-slip zones are represented by linearly concentrated rupture deformations. In contrast, near the termination of the fault the cluster fault deformation formed. Here, from north to south, there are radical changes in the palaeostress state. In the north-western sector (east of Selenga corridor) strike-slip faults, strike-slip faults with normal components and normal faults are dominated. For this sector the stress tensors of extensive, transtension and strike-slip regimes are typical. South-western sector is separated from the north-eastern one by massive Buren Nuruu ridge within which the active faults are not identified. In the south-western sector between the Orkhon and Tola rivers the cluster of NW thrusts and N-S strike-slip faults with reverse component are discovered. The faults are perfectly expressed by NW and N-S scarps in the relief. The most structures dip to the east and north-east. Holocene fault activity is demonstrated by the hanging river valleys and horizontal displacements with amplitudes

  12. Volcanic-ash hazard to aviation during the 2003-2004 eruptive activity of Anatahan volcano, Commonwealth of the Northern Mariana Islands

    USGS Publications Warehouse

    Guffanti, M.; Ewert, J.W.; Gallina, G.M.; Bluth, G.J.S.; Swanson, G.L.

    2005-01-01

    Within the Commonwealth of the Northern Mariana Islands (CNMI), Anatahan is one of nine active subaerial volcanoes that pose hazards to major air-traffic routes from airborne volcanic ash. The 2003-2004 eruptive activity of Anatahan volcano affected the region's aviation operations for 3 days in May 2003. On the first day of the eruption (10 May 2003), two international flights from Saipan to Japan were cancelled, and several flights implemented ash-avoidance procedures. On 13 May 2003, a high-altitude flight through volcanic gas was reported, with no perceptible damage to the aircraft. TOMS and MODIS analysis of satellite data strongly suggests that no significant ash and only minor amounts of SO2 were involved in the incident, consistent with crew observations. On 23 May 2003, airport operations were disrupted when tropical-cyclone winds dispersed ash to the south, dusting Saipan with light ashfall and causing flight cancellations there and at Guam 320 km south of the volcano. Operational (near-real-time) monitoring of ash clouds produced by Anatahan has been conducted since the first day of the eruption on 10 May 2003 by the Washington Volcanic Ash Advisory Center (VAAC). The VAAC was among the first groups outside of the immediate area of the volcano to detect and report on the unexpected eruption of Anatahan. After being contacted about an unusual cloud by National Weather Service forecasters in Guam at 1235 UTC on 10 May 2003, the VAAC analyzed GOES 9 images, confirming Anatahan as the likely source of an ash cloud and estimating that the eruption began at about 0730 UTC. The VAAC issued its first Volcanic Ash Advisory for Anatahan at 1300 UTC on 10 May 2003 more than 5 h after the start of the eruption, the delay reflecting the difficulty of detecting and confirming a surprise eruption at a remote volcano with no in situ real-time geophysical monitoring. The initial eruption plume reached 10.7-13.4 km (35,000-44,000 ft), well into jet cruise altitudes

  13. Volcanic-ash hazard to aviation during the 2003 2004 eruptive activity of Anatahan volcano, Commonwealth of the Northern Mariana Islands

    NASA Astrophysics Data System (ADS)

    Guffanti, Marianne; Ewert, John W.; Gallina, Gregory M.; Bluth, Gregg J. S.; Swanson, Grace L.

    2005-08-01

    Within the Commonwealth of the Northern Mariana Islands (CNMI), Anatahan is one of nine active subaerial volcanoes that pose hazards to major air-traffic routes from airborne volcanic ash. The 2003-2004 eruptive activity of Anatahan volcano affected the region's aviation operations for 3 days in May 2003. On the first day of the eruption (10 May 2003), two international flights from Saipan to Japan were cancelled, and several flights implemented ash-avoidance procedures. On 13 May 2003, a high-altitude flight through volcanic gas was reported, with no perceptible damage to the aircraft. TOMS and MODIS analysis of satellite data strongly suggests that no significant ash and only minor amounts of SO 2 were involved in the incident, consistent with crew observations. On 23 May 2003, airport operations were disrupted when tropical-cyclone winds dispersed ash to the south, dusting Saipan with light ashfall and causing flight cancellations there and at Guam 320 km south of the volcano. Operational (near-real-time) monitoring of ash clouds produced by Anatahan has been conducted since the first day of the eruption on 10 May 2003 by the Washington Volcanic Ash Advisory Center (VAAC). The VAAC was among the first groups outside of the immediate area of the volcano to detect and report on the unexpected eruption of Anatahan. After being contacted about an unusual cloud by National Weather Service forecasters in Guam at 1235 UTC on 10 May 2003, the VAAC analyzed GOES 9 images, confirming Anatahan as the likely source of an ash cloud and estimating that the eruption began at about 0730 UTC. The VAAC issued its first Volcanic Ash Advisory for Anatahan at 1300 UTC on 10 May 2003 more than 5 h after the start of the eruption, the delay reflecting the difficulty of detecting and confirming a surprise eruption at a remote volcano with no in situ real-time geophysical monitoring. The initial eruption plume reached 10.7-13.4 km (35,000-44,000 ft), well into jet cruise altitudes

  14. 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.

  15. 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...

  16. Tectonic events, continental intraplate volcanism, and mantle plume activity in northern Arabia: Constraints from geochemistry and Ar-Ar dating of Syrian lavas

    NASA Astrophysics Data System (ADS)

    Krienitz, M.-S.; Haase, K. M.; Mezger, K.; van den Bogaard, P.; Thiemann, V.; Shaikh-Mashail, M. A.

    2009-04-01

    New 40Ar/39Ar ages combined with chemical and Sr, Nd, and Pb isotope data for volcanic rocks from Syria along with published data of Syrian and Arabian lavas constrain the spatiotemporal evolution of volcanism, melting regime, and magmatic sources contributing to the volcanic activity in northern Arabia. Several volcanic phases occurred in different parts of Syria in the last 20 Ma that partly correlate with different tectonic events like displacements along the Dead Sea Fault system or slab break-off beneath the Bitlis suture zone, although the large volume of magmas and their composition suggest that hot mantle material caused volcanism. Low Ce/Pb (<20), Nb/Th (<10), and Sr, Nd, and Pb isotope variations of Syrian lavas indicate the role of crustal contamination in magma genesis, and contamination of magmas with up to 30% of continental crustal material can explain their 87Sr/86Sr. Fractionation-corrected major element compositions and REE ratios of uncontaminated lavas suggest a pressure-controlled melting regime in western Arabia that varies from shallow and high-degree melt formation in the south to increasingly deeper regions and lower extents of the beginning melting process northward. Temperature estimates of calculated primary, crustally uncontaminated Arabian lavas indicate their formation at elevated mantle temperatures (Texcess ˜ 100-200°C) being characteristic for their generation in a plume mantle region. The Sr, Nd, and Pb isotope systematic of crustally uncontaminated Syrian lavas reveal a sublithospheric and a mantle plume source involvement in their formation, whereas a (hydrous) lithospheric origin of lavas can be excluded on the basis of negative correlations between Ba/La and K/La. The characteristically high 206Pb/204Pb (˜19.5) of the mantle plume source can be explained by material entrainment associated with the Afar mantle plume. The Syrian volcanic rocks are generally younger than lavas from the southern Afro-Arabian region, indicating

  17. Affects of wastewater discharge from mining on soil heavy metal pollution and enzyme activities in northern Hunan province, Central South China

    NASA Astrophysics Data System (ADS)

    Jiang, Ying; Hu, Xue-Feng; Shu, Ying; Yan, Xiao-Juan; Luo, Fan

    2013-04-01

    Hunan province, Central South China, is rich in mineral resources and also a well-known nonferrous metal base in China. Mining and ore processing there, however, are mostly conducted in indigenous methods, and thus causing heavy metal pollution of abundant farmland. Situated in northern Hunan province, Y county has antimony, manganese, vanadium, and pyrite mines, but still belongs to a region of rice cultivation, of which, paddy fields make up 84.5% of the total farmland. Our investigations found that irrigation water is threatened by the release of mining wastewater in the county. For example, a stream used for irrigation turns dark-red after long-term receiving wastewater discharged from a pyrite company at HS Town of the county. Concentrations of Cu, Zn, Cd, Fe and Mn in the stream water reach 0.03 mg kg-1, 2.14 mg kg-1, 0.02 mg kg-1, 96.0 mg kg-1 and 11.5 mg kg-1, respectively; these in the paddy soils nearby are 67.3 mg kg-1, 297 mg kg-1, 4.0 mg kg-1, 33.1 mg g-1 and 463 mg kg-1 on average, respectively, with a maximum of Cd reaching 16.8 mg kg-1. Microbial biomass and activities are significantly reduced by metal toxicity in the soils. The counts of fungal, actinomycin and bacterial colonies in the polluted soils are 8.8×103 /g (Fresh soil), 4.9×105 /g (Fresh soil) and 6.4×105 /g (Fresh soil), respectively, which are only 4.68%, 10.3% and 20.9% of these in non-polluted soils in Y county, respectively. Likewise, the microbial biomass (MB) - C and MB - N of the polluted soils are only 36.8% and 50.3% of these in the non-polluted, respectively. The activities of dehydrogenase, urease, catalase, acid and neutral phosphatase and sucrase in the polluted soils are only 41.2%, 49.8%, 56.8%, 69.9%, 80.7% and 81.0% of these in the non-polluted, respectively. There are significant negative correlations between Cu, Zn and Cd contents and the activities of dehydrogenase and catalase, suggesting that the two enzymes are the most sensitive to heavy metal toxicity in the

  18. The northern Egyptian continental margin

    NASA Astrophysics Data System (ADS)

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

    2015-01-01

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

  19. Denitrification activity of aquifer material from two different sandy aquifers in northern Germany measured by laboratory incubation experiments in relation to influencing sediment parameters

    NASA Astrophysics Data System (ADS)

    Eschenbach, Wolfram; Well, Reinhard; Flessa, Heiner; Duijnisveld, Wilhelmus H. M.; Walther, Wolfgang

    2010-05-01

    In NO3¯ contaminated aquifers containing reduced compounds like organic carbon or sulfides, denitrification is an intense process. Its characterization is of interest because NO3¯ consumption improves water quality and N2O production can cause emission of this greenhouse gas to the atmosphere. Spatial distribution of NO3¯ and N2 produced by denitrification in groundwater (excess N2) reflects the NO3¯ input as well as cumulative denitrification during aquifer passage. The amount and spatial distribution of reduced compounds within denitrifying aquifers is not well known. Recent findings from parallel investigations on in situ denitrification and reactive compounds suggests that single-well 15N tracer tests might be suitable to characterize the stock of reduced compounds in aquifers (Konrad 2008), but an evaluation of this approach has not been realized until now. The overall objective of our studies is to measure the spatial and long-term dynamics of denitrification within two sandy aquifers in northern Germany. This includes measurement of the actually occurring denitrification process in the field by 15N-tracer experiments and incubation experiments with corresponding aquifer material. Beside the comparison of field and laboratory data concerning the denitrification activity, we also want to determine the long-term denitrification potential which is governed by the stock of reactive material within these sediments. Here we present the first results of these long-term incubation experiments to investigate the stock of reactive material present in these sediments and its denitrification activity. In these batch experiments we filled the aquifer material from different depths between two and 68 m below ground in glass bottles, supplemented it with K15NO3 solution, sealed the bottles airtight with rubber septa and flushed the headspace with pure N2. Afterward the bottles where stored in the dark by 10° C to obtain aquifer like conditions. The labeled

  20. New observations of the active deformation along the oblique collision/subduction boundary zone between the North American and Caribbean plates (northern Hispaniola offshore margin)

    NASA Astrophysics Data System (ADS)

    Carbó-Gorosabel, Andrés; Granja Bruña, José Luis; Rodríguez Zurrunero, Álvaro; Gómez de la Peña, Laura; Muñoz-Martín, Alfonso; Gómez Ballesteros, María; Gorosabel Araus, José Miguel; Espinosa, Salvador; Pazos, Anatonio; Catalán, Manuel; Yamil Rodríguez Asilis, Hector; Nuñez, José Luis; Muñoz, Santiago; ten Brink, Uri S.; Quijano, Jesús; Llanes Estrada, Pilar; Martín Dávila, José; Druet, María

    2015-04-01

    The Caribbean plate is moving relative to the North American plate at a rate of 20.0 ± 0.4 mm/y towards 074° ± 1°. This eastward motion has been taking place during most of the Cenozoic developing a 250 km-wide band of deformation, in which microplate and block tectonics take place. The eastward motion of the Hispaniola block is being impeded relative to the motion of the Caribbean plate's interior due to the collision with the Bahamas banks. This collision has resulted in the development of the Northern Hispaniola deformed belt along the northern Hispaniola offshore margin. A series of large (M6.2-M8.1) thrust earthquakes from 1943-1953, and two significant events in 1994 (M5.6) and 2003 (M6.4) occurred close to the city of Puerto Plata have been attributed to oblique collision/subduction of the North America plate and Bahamas banks beneath the northern Hispaniola. 300 km of 2D multi-channel seismic (MCS) reflection data and approximately 15000 km2 of high-resolution, systematic swath bathymetry data were recorded in the northern Hispaniola offshore margin as part of a larger survey carried out in November-December of 2013 aboard the Spanish R/V Sarmiento de Gamboa. MCS profiles were collected shooting a GI gun array (GGUN-II®) of 1750 ci. every 37.5 m and the signal recorded in a 3000 m-long streamer with 240 channels (Sentinel Sercel®). Differential GPS navigated high-resolution bathymetry data were collected using the hull-mounted Hydrosweep ATLAS DS echo-sounder system. Using new high-resolution multibeam bathymetry and MCS data, combined with previous 2D seismic data, we have studied the along- and across-strike variations of the geomorphology and shallower structure of the northern Hispaniola offshore margin. Here we present preliminary results focused on the identification and characterization of recent tectonic features in the region and provide well-defined targets to carry out future studies for seismic and tsunamigenic hazard assessment.

  1. 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

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. 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

  7. 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.

  8. Northern Trust Mines

    EPA Pesticide Factsheets

    The United States and the Navajo Nation entered into settlement agreements that provide funds to conduct investigations and any needed cleanup at 16 of the 46 priority mines, including six mines in the Northern Abandoned Uranium Mine Region.

  9. Active deformation in the Northern Walker Lane: a detailed geodetic study of the Mohawk Valley and Honey Lake/Warm Springs fault systems

    NASA Astrophysics Data System (ADS)

    Bormann, J. M.; Hammond, W. C.; Kreemer, C. W.; Blewitt, G.

    2012-12-01

    The Mohawk Valley and Honey Lake/Warm Springs faults are parallel, northwest striking, dextral fault systems separated by ~50 km in the westernmost part of the Northern Walker Lane. These two faults work as a cooperative pair to accommodate 3-5 mm/yr of the total 8 mm/yr of right-lateral deformation geodetically observed across the Northern Walker Lane, however it is unclear with fault is dominant. Geologic studies of the faults result in right-lateral slip rates of 1-2.5 mm/yr on the Honey Lake fault and a minimum of 0.3 mm/yr on the Mohawk Valley fault. In contrast, previous geodetic studies estimate slip rates of ~1 mm/yr on the Honey Lake fault and ~3 mm/yr on the Mohawk Valley fault. To reconcile the discrepancy between the distribution of slip on the faults and the differences between sums of the geologically and geodetically estimated slip rates, we use new GPS data to constrain an elastic block model developed specifically to study the Mohawk Valley and Honey Lake/Warm Springs fault systems. We present a dense GPS velocity solution (~10 km average station spacing) that incorporates new data from the semi-continuous Mobile Array of GPS for Nevada Transtension network (MAGNET, http://geodesy.unr.edu/networks) operated by the University of Nevada, Reno with continuous data from the EarthScope Plate Boundary Observatory and other networks. Data collected during the summer of 2012 bring many MAGNET GPS time series in the Northern Walker Lane to near 5 years in duration. The density of our velocity field and recent advances in data processing give us unprecedented precision in the measurement of contemporary deformation in the Northern Walker Lane. We use the velocity solution to solve for slip rates on the companion fault systems and explore the effects of block model geometry assumptions and tradeoffs. Our model predicts slip rates of 2.2±0.3 mm/yr for the Mohawk Valley fault and 1.1±0.2 mm/yr for the Honey Lake fault. Block model slip rate estimates are

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

    PubMed

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

    2016-08-26

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

  11. 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.

  12. 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.

  13. 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

  14. Common catabolic enzyme patterns in a microplankton community of the Humboldt Current System off northern and central-south Chile: Malate dehydrogenase activity as an index of water-column metabolism in an oxygen minimum zone

    NASA Astrophysics Data System (ADS)

    González, R. R.; Quiñones, R. A.

    2009-07-01

    An extensive subsurface oxygen minimum zone off northern and central-south Chile, associated with the Peru-Chile undercurrent, has important effects on the metabolism of the organisms inhabiting therein. Planktonic species deal with the hypoxic and anoxic environments by relying on biochemical as well as physiological processes related to their anaerobic metabolisms. Here we characterize, for the first time, the potential enzymatic activities involved in the aerobic and anaerobic energy production pathways of microplanktonic organisms (<100 μm), their relationship, and this relationship's association with the oxygen concentration and microplanktonic biomass in the oxygen minimum zone and adjacent areas of the Humboldt Current System water column. Our results demonstrate significant potential enzymatic activity of catabolic pathways in the oxygen minimum zone. Malate dehydrogenase had the highest oxidizing activity of nicotinamide adenine dinucleotide (reduced form) in the batch of catabolic enzymatic activities assayed, including potential pyruvate oxidoreductases activity, the electron transport system, and dissimilatory nitrate reductase. Malate dehydrogenase correlated significantly with almost all the enzymes analyzed within and above the oxygen minimum zone, and also with the oxygen concentration and microplankton biomass in the water column of the Humboldt Current System, especially in the oxygen minimum zone off Iquique. These results suggest a possible specific pattern for the catabolic activity of the microplanktonic realm associated with the oxygen minimum zone spread along the Humboldt Current System off Chile. We hypothesize that malate dehydrogenase activity could be an appropriate indicator of microplankton catabolism in the oxygen minimum zone and adjacent areas.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. 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

  1. 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.

  2. 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.

  3. Diapause-specific gene expression in the northern house mosquito, Culex pipiens L., identified by suppressive subtractive hybridization

    Technology Transfer Automated Retrieval System (TEKTRAN)

    In this study we probe the molecular events underpinning diapause observed in overwintering females of Culex pipiens. Using suppressive subtractive hybridization (SSH) we have identified 40 genes that are either upregulated or downregulated during this seasonal period of dormancy. Northern blot hybr...

  4. Northern Plains 'Crater'

    NASA Technical Reports Server (NTRS)

    2004-01-01

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

  5. Northern Plains of Mars

    NASA Technical Reports Server (NTRS)

    2004-01-01

    22 November 2004 This Mars Global Surveyor (MGS) Mars Orbiter Camera (MOC) image shows a typical view of the martian northern plains. Thousands of square kilometers of the northern middle and polar latitudes of Mars look similar to the scene in this image. In late spring and in summer, dust devils crisscross the northern plains, leaving a variety of dark streaks. The streaks do not survive from year to year, indicating their ephemeral nature. The circular features in this image, including the prominent bright circular feature near the bottom, are the locations of buried meteor impact craters. This image is located near 58.1oN, 207.6oW, and covers an area approximately 3 km (1.9 mi) wide. Sunlight illuminates the scene from the lower left.

  6. The Sanfandila earthquake sequence of 1998, Queretaro, Mexico: activation of an undocumented fault in the northern edge of central Trans-Mexican Volcanic Belt

    NASA Astrophysics Data System (ADS)

    Zúñiga, F. R.; Pacheco, J. F.; Guzmán-Speziale, M.; Aguirre-Díaz, G. J.; Espíndola, V. H.; Nava, E.

    2003-01-01

    A sequence of small earthquakes occurred in Central Mexico, at the northern edge of the Trans-Mexican Volcanic Belt (TMVB) in the State of Queretaro, during the first 3 months of 1998. Medium to large events in the continental regime of central Mexico are not common, but the seismic history of the region demonstrates that faults there are capable of generating destructive events. The sequence was analyzed using data from a temporary network with the goals of identifying the causative fault and its relation to regional tectonics. Employing a waveform inversion scheme adapted from a method used for regional studies, we found that the source mechanisms conform to the style of faulting (i.e. extension in the E-W direction) representative of the Taxco-San Miguel Allende Fault system. This system has been proposed as the southernmost extension of the Basin and Range (BR) Province. The spatial distribution of hypocenters and source mechanisms indicate that the seismogenic segment was a fault with an azimuth of approximately 334° with almost pure dip slip. Since events which occurred just south from this region show features which are consistent with TMVB tectonics (i.e. extension in an N-S direction), the sequence may mark the boundary between the TMVB and BR stress domains.

  7. Assessment of (226)Ra, (238)U, (232)Th, (137)Cs and (40)K activities from the northern coastline of Oman Sea (water and sediments).

    PubMed

    Darabi-Golestan, F; Hezarkhani, A; Zare, M R

    2017-02-27

    Water and sediment samples were collected from northern coast of Oman Sea covering from Goatr to Hormoz canyon seaport. Water and sediment quality assessment for naturally or anthropogenic radionuclides at Oman Sea as a main strategic golf for trade and transit, is an important issue. Correspondence analysis (CA) by R-mode analysis represents that sigma(T)-temperature-conductivity-O2 parameters are well-correlated with (226)Ra, (232)Th and (40)K. Accordingly Q-mode analysis revealed an indicator samples of ST13W for different radionuclides, ST03W for O2, and simultaneously ST34W-ST21W-ST08W-ST04W for sigma(T) parameters. The CA results overlapped with factor and cluster analysis results that explained 85.8% of total variance of water samples. Descriptive analysis of sediments indicates more significant variation than water samples. The (232)Th and (226)Ra generally showing that ST13D to ST25D sediments that restricted to Pi Bashk coastline are concentrated from (137)Cs and (40)K. By comparison with reference-values from Iran and other parts of the world, they are acceptable with respect to environmental and radioisotope hazards.

  8. Association of single nucleotide polymorphisms of APOBEC3G with susceptibility to HIV-1 infection and disease progression among men engaging in homosexual activity in northern China.

    PubMed

    Li, Qiuyan; Qiao, Yuandong; Zhang, Guangfa; He, Ning; Zhang, Xuelong; Jia, Xueyuan; Sun, Haiming; Wang, Chuntao; Xu, Lidan

    2017-01-01

    Men who have sex with men (MSM) are at high risk of HIV infection. The APOBEC3G (apolipoprotein B mRNA editing catalytic polypeptide 3G) protein is a component of innate antiviral immunity that inhibits HIV-1 replication. In the present study, a total of 483 HIV-1 seropositive men and 493 HIV-1 seronegative men were selected to investigate the association between single nucleotide polymorphisms (SNPs) of the APOBEC3G gene and susceptibility to HIV-1 infection and AIDS progression among MSM residing in northern China. Genotyping of four SNPs (rs5757465, rs3736685, rs8177832, and rs2899313) of the APOBEC3G was performed using the SNPscan™ Kit, while the rs2294367 polymorphism was genotyped using the SNaPshot multiplex system. Our results disclosed no association between the SNPs of APOBEC3G and susceptibility to HIV-1, or effects of these polymorphisms on the CD4(+) T cell count or clinical phase of disease. A meta-analysis of 1624 men with HIV-1 infection and 1523 controls suggested that the association between rs8177832 and susceptibility was not significant. However, we observed a trend towards association with HIV-1 infection for haplotype TTACA (p = 0.082). The potential role of variants of APOBEC3G in HIV-1/AIDS warrants further investigation.

  9. Detection of haemoparasites in cattle by reverse line blot hybridisation with a note on the distribution of ticks in Sicily.

    PubMed

    Georges, K; Loria, G R; Riili, S; Greco, A; Caracappa, S; Jongejan, F; Sparagano, O

    2001-08-31

    A reverse line blot hybridisation (RLB) of 21 oligonucleotides with polymerase chain reaction (PCR) amplified regions of 16S rRNA (Ehrlichia/Anaplasma group) or 18S rRNA (Babesia/Theileria group) genes of haemoparasites detected Theileria annulata, T. buffeli/orientalis, Babesia bovis, B. bigemina, B. divergens, Ehrlichia bovis, Anaplasma marginale, A. centrale and unknown species within the Rickettsia tribe.A very high prevalence of mixed infections was detected, which indicated that animals infected with Babesia spp. were also infected with Theileria spp. and/or Anaplasma spp. The tick distribution appeared to be seasonal with Hyalomma marginatum as the most frequently observed tick and Boophilus annulatus and Ixodes ricinus as the least frequently observed ticks. Other species identified in the 818 ticks collected during the five sampling periods between April 1998 and November 1999 included H. lusitanicum, Rhipicephalus sanguineus group, R. bursa, Dermacentor marginatus, Haemaphysalis punctata, B. annulatus and I. ricinus.

  10. Eighteen Years of Molecular Genotyping the Hemophilia Inversion Hotspot: From Southern Blot to Inverse Shifting-PCR

    PubMed Central

    Rossetti, Liliana C.; Radic, Claudia P.; Abelleyro, Miguel M.; Larripa, Irene B.; De Brasi, Carlos D.

    2011-01-01

    The factor VIII gene (F8) intron 22 inversion (Inv22) is a paradigmatic duplicon-mediated rearrangement, found in about one half of patients with severe hemophilia A worldwide. The identification of this prevalent cause of hemophilia was delayed for nine years after the F8 characterization in 1984. The aim of this review is to present the wide diversity of practical approaches that have been developed for genotyping the Inv22 (and related int22h rearrangements) since discovery in 1993. The sequence— Southern blot, long distance-PCR and inverse shifting-PCR—for Inv22 genotyping is an interesting example of scientific ingenuity and evolution in order to resolve challenging molecular diagnostic problems. PMID:22072947

  11. Detection of human cytomegalovirus by slot-blot hybridization assay employing oligo-primed /sup 32/P-labelled probe

    SciTech Connect

    Agha, S.A.; Coleman, J.C.; Selwyn, S.; Mahmound, L.A.; Abd-Elaal, A.M.; Archard, L.C.

    1988-12-01

    A /sup 32/P-labelled Hind III-0 DNA fragment (nine Kilobases; Kb) from human cytomegalovirus AD-169 (HCMV) was used in slot-blot hybridization assay for the detection of HCMV in clinical samples. The results obtained with DNA hybridization assay (DNA HA) were compared with virus isolation using conventional tube cell culture (CTC) and centrifugation vial culture (CVC), immunofluorescence (IF), and complement fixation test (CFT). Of 15 CTC-positive samples, 13 were positive with DNA HA (sensitivity 86.7%). Also, 14 additional samples were DNA HA-positive but CTC-negative. CVC and/or IF confirmed the diagnosis in nine of 14; the remaining five samples were from three patients who showed fourfold rising antibody titre by CFT. Although DNA HA using /sup 32/P-labelled probes is relatively cumbersome and expensive, it is a valuable test for quantitation of viral shedding in patients with HCMV infections who may benefit from antiviral therapy.

  12. 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

  13. 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.

  14. 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.

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

    SciTech Connect

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

    1988-08-25

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

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

    SciTech Connect

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

    1994-09-01

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

  17. Northern Arizona University

    ERIC Educational Resources Information Center

    Butcher, Michael F.; Saltonstall, Margot; Bickel, Sarah; Brandel, Rick

    2009-01-01

    Northern Arizona University (NAU) is a public university nestled below the San Francisco Peaks in Flagstaff, Arizona. It enrolls more than 21,000 undergraduate and graduate students at its main campus in Flagstaff, through its 35 statewide sites, and via online program offerings. Within the university organizational system, Student Affairs has a…

  18. 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.

  19. Northern Pintail (Anas acuta)

    USGS Publications Warehouse

    Austin, J.E.; Miller, M.R.; Poole, A.; Gill, F.

    1995-01-01

    The Northern Pintail is a medium-sized dabbling duck of slender, elegant lines and conservative plumage coloration. It is circumpolar in distribution and abundant in North America, with core nesting habitat in Alaska and the Prairie Pothole Region of southern Canada and the northern Great Plains. Breeders favor shallow wetlands interspersed throughout prairie grasslands or arctic tundra. An early fall migrant, the species arrives on wintering areas beginning in August, after wing molt, often forming large roosting and feeding flocks on open, shallow wetlands and flooded agricultural fields. The birds consume grains, marsh plant seeds, and aquatic invertebrates throughout the fall and winter. Northern Pintails are among the earliest nesting ducks in North America, beginning shortly after ice-out in many northern areas. Individuals form new pair bonds each winter but are highly promiscuous during the nesting season, with mated and unmated males often involved in vigorous, acrobatic Pursuit Flights. Annual nest success and productivity vary with water conditions, predation, and weather. Females build nests on the ground, often long distances from water. Only the female incubates; her mate leaves shortly after incubation begins. Ducklings hatch together in one day, follow the female to water after a day in the nest, and fledge by July or August. Adults and ducklings consume mainly aquatic invertebrates during the breeding season. Predators and farming operations destroy many thousands of Northern Pintail nests annually; farming has also greatly reduced the amount of quality nesting cover available. Winter habitats are threatened by water shortages, agricultural development, contamination, and urbanization. Periods of extended drought in prairie nesting regions have caused dramatic population declines, usually followed by periods of recovery. Over the long term, however, the continental population of Northern Pintails has declined significantly from 6 million birds in

  20. Patterns of extracellular enzyme activities and microbial metabolism in an Arctic fjord of Svalbard and in the northern Gulf of Mexico: contrasts in carbon processing by pelagic microbial communities.

    PubMed

    Arnosti, Carol; Steen, Andrew D

    2013-01-01

    The microbial community composition of polar and temperate ocean waters differs substantially, but the potential functional consequences of these differences are largely unexplored. We measured bacterial production, glucose metabolism, and the abilities of microbial communities to hydrolyze a range of polysaccharides in an Arctic fjord of Svalbard (Smeerenburg Fjord), and thus to initiate remineralization of high-molecular weight organic matter. We compared these data with similar measurements previously carried out in the northern Gulf of Mexico in order to investigate whether differences in the spectrum of enzyme activities measurable in Arctic and temperate environments are reflected in "downstream" aspects of microbial metabolism (metabolism of monomers and biomass production). Only four of six polysaccharide substrates were hydrolyzed in Smeerenburg Fjord; all were hydrolyzed in the upper water column of the Gulf. These patterns are consistent on an interannual basis. Bacterial protein production was comparable at both locations, but the pathways of glucose utilization differed. Glucose incorporation rate constants were comparatively higher in Svalbard, but glucose respiration rate constants were higher in surface waters of the Gulf. As a result, at the time of sampling ca. 75% of the glucose was incorporated into biomass in Svalbard, but in the northern Gulf of Mexico most of the glucose was respired to CO2. A limited range of enzyme activities is therefore not a sign of a dormant community or one unable to further process substrates resulting from extracellular enzymatic hydrolysis. The ultimate fate of carbohydrates in marine waters, however, is strongly dependent upon the specific capabilities of heterotrophic microbial communities in these disparate environments.

  1. Patterns of extracellular enzyme activities and microbial metabolism in an Arctic fjord of Svalbard and in the northern Gulf of Mexico: contrasts in carbon processing by pelagic microbial communities

    PubMed Central

    Arnosti, Carol; Steen, Andrew D.

    2013-01-01

    The microbial community composition of polar and temperate ocean waters differs substantially, but the potential functional consequences of these differences are largely unexplored. We measured bacterial production, glucose metabolism, and the abilities of microbial communities to hydrolyze a range of polysaccharides in an Arctic fjord of Svalbard (Smeerenburg Fjord), and thus to initiate remineralization of high-molecular weight organic matter. We compared these data with similar measurements previously carried out in the northern Gulf of Mexico in order to investigate whether differences in the spectrum of enzyme activities measurable in Arctic and temperate environments are reflected in “downstream” aspects of microbial metabolism (metabolism of monomers and biomass production). Only four of six polysaccharide substrates were hydrolyzed in Smeerenburg Fjord; all were hydrolyzed in the upper water column of the Gulf. These patterns are consistent on an interannual basis. Bacterial protein production was comparable at both locations, but the pathways of glucose utilization differed. Glucose incorporation rate constants were comparatively higher in Svalbard, but glucose respiration rate constants were higher in surface waters of the Gulf. As a result, at the time of sampling ca. 75% of the glucose was incorporated into biomass in Svalbard, but in the northern Gulf of Mexico most of the glucose was respired to CO2. A limited range of enzyme activities is therefore not a sign of a dormant community or one unable to further process substrates resulting from extracellular enzymatic hydrolysis. The ultimate fate of carbohydrates in marine waters, however, is strongly dependent upon the specific capabilities of heterotrophic microbial communities in these disparate environments. PMID:24198812

  2. Changes in phenylalanine ammonia-lyase activity and gene expression during storage of asparagus spears.

    PubMed

    Bhowmik, Pankaj K; Matsui, Toshiyuki

    2005-01-01

    A cDNA clone coding phenylalanine ammonia-lyase (PAL) was isolated from a cDNA library prepared from asparagus spears (Asparagus officinalis L. cv. Welcome) using the reverse transcription-polymerase chain reaction (RT-PCR). The partial cDNA clone encoded an mRNA of 527 bp and the derived amino acid sequence was found highly homologous to PAL from rice, maize and barley. Northern blot analysis showed an increase of pAS-PAL mRNA until 24 h at 20 degrees C, which coincided well with PAL activity and fiber development, suggesting that the increase is a response to the wounding associated with harvest.

  3. Geology of the Ivanhoe Hg-Au district, northern Nevada: Influence of Miocene volcanism, lakes, and active faulting on epithermal mineralization

    USGS Publications Warehouse

    Wallace, A.R.

    2003-01-01

    The mercury-gold deposits of the Ivanhoe mining district in northern Nevada formed when middle Miocene rhyolitic volcanism and high-angle faulting disrupted a shallow lacustrine environment. Sinter and replacement mercury deposits formed at and near the paleosurface, and disseminated gold deposits and high-grade gold-silver veins formed beneath the hot spring deposits. The lacustrine environment provided abundant meteoric water; the rhyolites heated the water; and the faults, flow units, and lakebeds provided fluid pathways for the hydrothermal fluids. A shallow lake began to develop in the Ivanhoe area about 16.5 Ma. The lake progressively expanded and covered the entire area with fine-grained lacustrine sediments. Lacustrine sedimentation continued to at least 14.4 Ma, and periodic fluctuations in the size and extent of the lake may have been responses to both climate and nearby volcanism. The eruption of rhyolite and andesite flows and domes periodically disrupted the lacustrine environment and produced interfingered flows and lake sediments. The major pulse of rhyolitic volcanism took place between 15.16 ± 0.05 and 14.92 ± 0.05 Ma. High-angle faulting began in the basement about 15.2 Ma, penetrated to and disrupted the paleosurface after 15.10 ± 0.06 Ma, and largely ceased by 14.92 ± 0.05 Ma. Ground motion related to both faulting and volcanism created debris flows and soft-sediment deformation in the lakebeds. Mercury-gold mineralization was coeval with rhyolite volcanism and high-angle faulting, and it took place about 15.2 to 14.9 Ma. At and near the paleosurface, hydrothermal fluids migrated through tuffaceous sediments above relatively impermeable volcanic and Paleozoic units, creating chalcedonic, cinnabar-bearing replacement bodies and sinters. Disseminated gold was deposited in sedimentary and volcanic rocks beneath the mercury deposits, although the hydrologic path between the two ore types is unclear. Higher-grade gold-silver deposits formed in

  4. Assessment of Ficam VC (Bendiocarb) Residual Activity on Different Wall Surfaces for Control of Anopheles gambiae s.s. (Diptera: Culicidae) in Northern Uganda.

    PubMed

    Kirunda, James; Okello-Onen, Joseph; Opiyo, Elizabeth A; Rwakimari, J B; de Alwis, Ranjith; Okia, Michael; Ambayo, Denis; Abola, Benard; Hoel, David F

    2017-03-24

    Insecticide decay rate on different wall surfaces is of importance to indoor residual spray (IRS) programs used as a malaria control intervention. Past IRS operations showed increasing populations of endophilic malaria vectors resting on indoor surfaces from various sites in Uganda following use of Ficam VC (bendiocarb) insecticide; variability of insecticide life was believed to be primarily due to wall surface type. Bendiocarb longevity was tested in the northern Uganda districts of Amuru, Apac, and Pader to assess its residual efficacy on three commonly encountered wall surfaces. Wall types included mud and wattle, plain brick, and painted plaster. A susceptible mosquito strain (Anopheles gambiae Kisumu) was used in all trials. Nine houses in each of the three districts were set with three test cones and one control cone per house, divided evenly among the three wall surfaces. Bioassays were run monthly through 6 mo. Painted plastered surfaces produced 100% mortality (at 24 h) through 6 mo. Plain brick surfaces killed 100% of test mosquitoes through 4 mo, while mud and wattle wall surfaces produced a 98% mortality rate at 3 mo post spray. The KD60 (knockdown at 60 min) for painted plastered surfaces was 100% for 6 mo, plain brick surface KD60 was 80% at 6 mo, and the mud and wattle surface KD60 was >80% at 3 mo. There was a significant effect on Ficam VC longevity by wall type and evidence of a relationship between test period and wall type on the KD60.

  5. Plague studies in California: a review of long-term disease activity, flea-host relationships and plague ecology in the coniferous forests of the Southern Cascades and northern Sierra Nevada mountains.

    PubMed

    Smith, Charles R; Tucker, James R; Wilson, Barbara A; Clover, James R

    2010-06-01

    We review 28 years of long-term surveillance (1970-1997) for plague activity among wild rodents from ten locations within three coniferous forest habitat types in the northern Sierra Nevada and the Southern Cascade mountains of northeastern California. We identify rodent hosts and their fleas and document long-term plague activity in each habitat type. The highest seroprevalence for Yersinia pestis occurred in the chipmunks, Tamias senex and T. quadrimaculatus, and the pine squirrel, Tamiasciurus douglasii. The most commonly infected fleas were Ceratophyllus ciliatus and Eumolpianus eumolpi from chipmunks and Oropsylla montana and O. idahoensis from ground squirrels. Serological surveillance demonstrated that populations of T. senex, T. quadrimaculatus and T. douglasii are moderately resistant to plague, survive infection, and are, therefore, good sentinels for plague activity. Recaptured T. senex and T. quadrimaculatus showed persistence of plague antibodies and evidence of re-infection over a two year period. These rodent species, their fleas, and the ecological factors common to the coniferous forest habitats likely promote the maintenance of plague foci in northeastern California.

  6. 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.

  7. Northern Plains Patterns

    NASA Technical Reports Server (NTRS)

    2003-01-01

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

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

  8. Dermatophytosis in northern Africa.

    PubMed

    Nweze, E I; Eke, I

    2016-03-01

    Infections caused by dermatophytes are a global problem and a major public health burden in the world today. In Africa, especially in the northern geographical zone, dermatophytic infections are being reported at an alarming rate. This is mostly because of some local but unique cultural practices, socioeconomic and environmental conditions, lack of reliable diagnostic personnel and facilities and ineffective treatment. Interestingly, the pathogen spectrum and the clinical manifestation are most times different from what is seen in other continents. Several epidemiological studies have been performed on the incidence and aetiology of dermatophytoses in northern Africa. However, there is currently no review article with up-to-date information on the relevant findings reported so far in this region. This information is necessary for clinicians who treat dermatophytic infections all over the world since agents of dermatophytes are no longer restricted because of the rapid mobility of humans from one part of the world to another. Moreover, the epidemiology of dermatophytoses is known to change over time, thus requiring the update of information from time to time. A review of relevant studies published on dermatophytoses in northern Africa is presented. This covers all of old Sudan, Algeria, Egypt, Libya, Tunisia and Morocco.

  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. Comparison of germ line minisatellite mutation detection at the CEB1 locus by Southern blotting and PCR amplification.

    PubMed

    Taylor, Malcolm; Cieslak, Marcin; Rees, Gwen S; Oojageer, Anthony; Leith, Cheryl; Bristow, Claire; Tawn, E Janet; Winther, Jeanette F; Boice, John D

    2010-07-01

    Identification of de novo minisatellite mutations in the offspring of parents exposed to mutagenic agents offers a potentially sensitive measure of germ line genetic events induced by ionizing radiation and genotoxic chemicals. Germ line minisatellite mutations (GMM) are usually detected by hybridizing Southern blots of unamplified size-fractionated genomic DNA with minisatellite probes. However, this consumes a relatively large amount of DNA, requires several steps and may lack sensitivity. We have developed a polymerase chain reaction (PCR)-based GMM assay, which we applied to the hypermutable minisatellite, CEB1. Here, we compare the sensitivity and specificity of this assay with the conventional Southern hybridization method using DNA from 10 spouse pairs, one parent of each pair being a survivor of cancer in childhood, and their 20 offspring. We report that both methods have similar specificity but that the PCR method uses 250 times less DNA, has fewer steps and is better at detecting GMM with single repeats provided that specific guidelines for allele sizing are followed. The PCR GMM method is easier to apply to families where the amount of offspring DNA sample is limited.

  16. 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.

  17. Climate impacts on northern Canada: regional background.

    PubMed

    Prowse, Terry D; Furgal, Chris; Bonsal, Barrie R; Peters, Daniel L

    2009-07-01

    Understanding the implications of climate change on northern Canada requires a background about the size and diversity of its human and biogeophysical systems. Occupying an area of almost 40% of Canada, with one-third of this contained in Arctic islands, Canada's northern territories consist of a diversity of physical environments unrivaled around the circumpolar north. Major ecozones composed of a range of landforms, climate, vegetation, and wildlife include: Arctic, boreal and taiga cordillera; boreal and taiga plains; taiga shield; and northern and southern Arctic. Although generally characterized by a cold climate, there is an enormous range in air temperature with mean annual values being as high as -5 degrees C in the south to as low as -20 degrees C in the high Arctic islands. A similar contrast characterizes precipitation, which can be > 700 mm y(-1) in some southern alpine regions to as low as 50 mm y(-1) over islands of the high Arctic. Major freshwater resources are found within most northern ecozones, varying from large glaciers or ice caps and lakes to extensive wetlands and peat lands. Most of the North's renewable water, however, is found within its major river networks and originates in more southerly headwaters. Ice covers characterize the freshwater systems for multiple months of the year while permafrost prevails in various forms, dominating the terrestrial landscape. The marine environment, which envelops the Canadian Arctic Archipelago, is dominated by seasonal to multiyear sea ice often several meters thick that plays a key role in the regional climate. Almost two-thirds of northern Canadian communities are located along coastlines with the entire population being just over 100 000. Most recent population growth has been dominated by an expansion of nonaboriginals, primarily the result of resource development and the growth of public administration. The economies of northern communities, however, remain quite mixed with traditional land

  18. [Social representations concerning dengue, dengue vectors, and control activities among residents of São Sebastião on the northern coast of São Paulo State, Brazil].

    PubMed

    Lefèvre, Ana Maria Cavalcanti; Ribeiro, Andressa Francisca; Marques, Gisela Rita de Alvarenga Monteiro; Serpa, Lígia Leandro Nunes; Lefèvre, Fernando

    2007-07-01

    This study sought to identify people's knowledge on dengue and its vector biology, aimed at promoting a communications channel between technical and lay reasoning in order to foster community involvement in dengue and dengue vector control activities. A survey was conducted in an Aedes aegypti-infested area with dengue transmission in São Sebastião on the northern coast of São Paulo State, Brazil. One hundred interviews were held, with five open questions on topics related to dengue and vector control. Collective Subject Discourse methodology was used in the analysis. People were not able to properly identify the kinds of accumulated water sources that serve as breeding places for mosquitoes and were unaware of the egg phase in vector development. There was inadequate awareness of vector biology and a need for greater government-community integration. Educational activities should incorporate the study results as insight for improving the social efficiency and efficacy of joint actions to fight dengue and control the mosquito vector.

  19. Using of Remote Sensing Techniques for Monitoring the Earthquakes Activities Along the Northern Part of the Syrian Rift System (LEFT-LATERAL),SYRIA

    NASA Astrophysics Data System (ADS)

    Dalati, Moutaz

    Earthquake mitigation can be achieved with a better knowledge of a region's infra-and substructures. High resolution Remote Sensing data can play a significant role to implement Geological mapping and it is essential to learn about the tectonic setting of a region. It is an effective method to identify active faults from different sources of Remote Sensing and compare the capability of some satellite sensors in active faults survey. In this paper, it was discussed a few digital image processing approaches to be used for enhancement and feature extraction related to faults. Those methods include band ratio, filtering and texture statistics . The experimental results show that multi-spectral images have great potentials in large scale active faults investigation. It has also got satisfied results when deal with invisible faults. Active Faults have distinct features in satellite images. Usually, there are obvious straight lines, circular structures and other distinct patterns along the faults locations. Remotely Sensed imagery Landsat ETM and SPOT XS /PAN are often used in active faults mapping. Moderate and high resolution satellite images are the best choice, because in low resolution images, the faults features may not be visible in most cases. The area under study is located Northwest of Syria that is part of one of the very active deformation belt on the Earth today. This area and the western part of Syria are located along the great rift system (Left-Lateral or African- Syrian Rift System). Those areas are tectonically active and caused a lot of seismically events. The AL-Ghab graben complex is situated within this wide area of Cenozoic deformation. The system formed, initially, as a result of the break up of the Arabian plate from the African plate. This action indicates that these sites are active and in a continual movement. In addition to that, the statistic analysis of Thematic Mapper data and the features from a digital elevation model ( DEM )produced from

  20. Seasonal Variation in Soil Microbial Biomass, Bacterial Community Composition and Extracellular Enzyme Activity in Relation to Soil Respiration in a Northern Great Plains Grassland

    NASA Astrophysics Data System (ADS)

    Wilton, E.; Flanagan, L. B.

    2014-12-01

    Soil respiration rate is affected by seasonal changes in temperature and moisture, but is this a direct effect on soil metabolism or an indirect effect caused by changes in microbial biomass, bacterial community composition and substrate availability? In order to address this question, we compared continuous measurements of soil and plant CO2 exchange made with an automatic chamber system to analyses conducted on replicate soil samples collected on four dates during June-August. Microbial biomass was estimated from substrate-induced respiration rate, bacterial community composition was determined by 16S rRNA amplicon pyrosequencing, and β-1,4-N-acetylglucosaminidase (NAGase) and phenol oxidase enzyme activities were assayed fluorometrically or by absorbance measurements, respectively. Soil microbial biomass declined from June to August in strong correlation with a progressive decline in soil moisture during this time period. Soil bacterial species richness and alpha diversity showed no significant seasonal change. However, bacterial community composition showed a progressive shift over time as measured by Bray-Curtis dissimilarity. In particular, the change in community composition was associated with increasing relative abundance in the alpha and delta classes, and declining abundance of the beta and gamma classes of the Proteobacteria phylum during June-August. NAGase showed a progressive seasonal decline in potential activity that was correlated with microbial biomass and seasonal changes in soil moisture. In contrast, phenol oxidase showed highest potential activity in mid-July near the time of peak soil respiration and ecosystem photosynthesis, which may represent a time of high input of carbon exudates into the soil from plant roots. This input of exudates may stimulate the activity of phenol oxidase, a lignolytic enzyme involved in the breakdown of soil organic matter. These analyses indicated that seasonal change in soil respiration is a complex

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

    PubMed Central

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

    2013-01-01

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

  2. Molecular Detection and Genotyping of Male-Specific Coliphages by Reverse Transcription-PCR and Reverse Line Blot Hybridization

    PubMed Central

    Vinjé, Jan; Oudejans, Sjon J. G.; Stewart, Jill R.; Sobsey, Mark D.; Long, Sharon C.

    2004-01-01

    In recent years, there has been increased interest in the use of male-specific or F+ coliphages as indicators of microbial inputs to source waters. Sero- or genotyping of these coliphages can also be used for microbial source tracking (MST). Among the male-specific coliphages, the F+ RNA (FRNA) viruses are well studied, while little is known about the F+ DNA (FDNA) viruses. We have developed a reverse line blot hybridization (RLB) assay which allows for the simultaneous detection and genotyping of both FRNA as well as FDNA coliphages. These assays included a novel generic duplex reverse transcription-PCR (RT-PCR) assay for FRNA viruses as well as a generic PCR for FDNA viruses. The RT-PCR assays were validated by using 190 field and prototype strains. Subsequent DNA sequencing and phylogenetic analyses of RT-PCR products revealed the classification of six different FRNA clusters, including the well-established subgroups I through IV, and three different FDNA clusters, including one (CH) not previously described. Within the leviviruses, a potentially new subgroup (called JS) including strains having more than 40% nucleotide sequence diversity with the known levivirus subgroups (MS2 and GA) was identified. We designed subgroup-specific oligonucleotides that were able to genotype all nine (six FRNA, three FDNA) different clusters. Application of the method to a panel of 351 enriched phage samples from animal feces and wastewater, including known prototype strains (MS2, GA, Qβ, M11, FI, and SP for FRNA and M13, f1, and fd for FDNA), resulted in successful genotyping of 348 (99%) of the samples. In summary, we developed a novel method for standardized genotyping of F+ coliphages as a useful tool for large-scale MST studies. PMID:15466543

  3. 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.

  4. Reconstructing the ecological impacts of eight decades of mining, metallurgical, and municipal activities on a small boreal lake in northern Canada.

    PubMed

    Doig, Lorne E; Schiffer, Stephanie T; Liber, Karsten

    2015-07-01

    As a result of long-term metal mining and metallurgical activities, the sediment of Ross Lake (Flin Flon, MB, Canada) is highly contaminated with metals and other elements. Although the effluents likely were discharged into Ross Lake as early as the late 1920s, lake biophysical data were not collected until 1973, more than 4 decades after the onset of mining and municipal activities. The early influence of these activities on the ecology of Ross Lake is unknown, as are the effects of improvements to metallurgical effluent quality and discontinuation of municipal wastewater discharge into the lake's north basin. To address this knowledge gap, analyses typical of paleolimnological investigations were applied to cores of sediment collected in 2009 from the south basin of Ross Lake. Stratigraphic analyses of physicochemical sediment characteristics (e.g., the concentrations of metals and other elements, organic C, total N, and δ(13)C and δ(15)N values) and subfossil remains (diatoms, Chironomidae, Chaoborus, and Cladocera) were used to infer historical biological and chemical changes in Ross Lake. With the onset of mining activities, concentrations of various elements (e.g., As, Cr, Cu, Zn, and Se) increased dramatically in the sediment profile, eventually declining with improved tailings management. Nevertheless, concentrations of metals in recent sediments remain elevated compared with pre-industrial sediments. Constrained cluster analyses demonstrated distinct pre-industrial and postindustrial communities for both the diatoms and chironomids. The biodiversity of the postindustrial diatom assemblages were much reduced compared with the pre-industrial assemblages. The postindustrial chironomid assemblage was dominated by Chironomus and to a lesser extent by Procladius, suggesting that Ross Lake became a degraded environment. Abundances of Cladocera and Chaoborus were severely reduced in the postindustrial era, likely because of metals toxicity. Overall, improvements

  5. 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.

  6. Short-term planetary-scale interactions over the tropics and midlatitudes during northern winter. I - Contrasts between active and inactive periods

    NASA Technical Reports Server (NTRS)

    Chang, C.-P.; Lau, K. M.

    1982-01-01

    Objectively analyzed 200 mb winds of four winters are used to study the short-term (several days) teleconnections between planetary-scale circulation components over the monsoon region. The composited structures suggest that during very actively northeasterly monsoon (surge) periods, the midlatitude and tropical circulation components vary in a coherent way. The jet streak and local Hadley circulation over East Asia, the divergent flow over the maritime continent, and the equatorial Walker circulations over the Pacific and Indian Oceans all strengthen steadily, while the secondary jet streak over West Asia weakens. During inactive (break) periods the midlatitude circulation components all exhibit reversed changes, while the variations in the tropics are less coherent, although still showing reversed tendencies. The results basically verify the short-term teleconnection model proposed by Chang and Lau (1980), but more complex processes in the acceleration of the midlatitude jet streaks are indicated. In addition, the contrast between the very active and break monsoon periods suggests the relative importance of tropical versus midlatitudinal forcings in different regions of the monsoon circulation.

  7. [Impact of automobile exhaust on membrane lipid peroxidation and protective enzyme activities in seedlings foliage of four northern broadleaved tree species].

    PubMed

    Ma, Shuhua; Wang, Qingcheng; Li, Yacang

    2004-12-01

    By means of fumigating one-year-old seedlings in open top chambers, this paper studied the impact of automobile exhaust on the pH value, relative conductivity, malondialdehyde (MDA) and chlorophyll contents, superoxide dismutase (SOD) and peroxidase (POD) activities, and ascorbic acid (ASA) content in the seedlings foliage of four tree species, Acer mono, Malus baccata, Prunus ussuriensis, and Acer ginnala. During the fumigation, the seedlings were exposed to the same exhaust gas concentration (25 microg x m(-3), indicated by the NO2 concentration in exhaust) for different durations (1, 3, 5, 7 d), and to different concentrations (40, 60, 80, 100 microg NO2 x m(-3)) for same duration (2 h). The results showed that the pH value and the chlorophyll and ascorbic acid (ASA) contents decreased, whereas the relative conductivity, malondialdehyde (MDA) content, and superoxide dismutase (SOD) and peroxidase (POD) activities increased with increasing fumigation duration and exhaust concentration. Obvious interspecies variations in term of physiological features were found. After treated 7 days with 25 microg NO2 x m(-3) and treated 2 h with 100 microg NO2 x m(-3), only a 1.5% and 2.7% decrease of cell juice pH was found in A. ginnala, respectively, compared to the control. The corresponding data for P. ussuriensis was 9.42% and 13.89%, followed by M. baccata. The chlorophyll content of A. mono, A. ginnala, M. baccata and P. ussuriensis was 83.0%, 71.3%, 68.7% and 54.9%, respectively of the control after 7 days treated with 25 microg NO2 x m(-3), and the corresponding data under 100 microg NO2 x m(-3) treatment was 60.2%, 73.1%, 43.4% and 51.2%, respectively. The decrease of ASA content and Acer ginnala was less in A. mono than in M. baccata and P. ussuriensis. The relative conductivity and MDA content of A. mono increased respectively by 68.1% and 52.5% in compared with control, while those of A. ginnala had the least increment. As for the 100 microg NO2 x m(-3) treatment

  8. 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.

  9. 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

  10. Enzyme-linked immunoelectrotransfer blot analysis of a cryptosporidiosis outbreak on a United States Coast Guard cutter.

    PubMed

    Moss, D M; Bennett, S N; Arrowood, M J; Wahlquist, S P; Lammie, P J

    1998-01-01

    Symptoms consistent with an outbreak of cryptosporidiosis (diarrhea, vomiting, nausea, and abdominal cramps) occurred on a U.S. Coast Guard cutter within 0-18 days after the cutter filled its tanks with Milwaukee, Wisconsin city water in March 1993. At three-weeks postdocking (PD), the suspected water was removed, and serum samples and stool specimens were collected from 47 of the 58 crew members, as well as questionnaire data on their water consumption and symptoms aboard the cutter. At 10-weeks PD and/or at 28-weeks PD, additional serum specimens were collected. Intensitometric data from enzyme-linked immunoelectrotransfer blot (EITB) were obtained on IgA responses to a 17-kD antigen group, IgM responses to a 27-kD antigen group, and IgG responses to 27-, 17-, and 15-kD antigen groups extracted from oocysts. In addition, IgG responses to crude oocyst antigens were obtained by ELISA. Based on reported symptoms, EITB results, and stool examination, the crew members were classified as confirmed (10), probable (10), suspected (22), and noncases (16). Of the 10 confirmed cases (all symptomatic) and the 10 probable cases (eight symptomatic) whose stools were positive and negative, respectively, for Cryptosporidium oocysts by microscopy, all showed changes in EITB intensities to the antigen groups and were considered EITB positive. The remaining 38 crew members, 22 suspected cases (all symptomatic), and 16 noncases (all asymptomatic), if tested, had negative stool examinations and were considered EITB negative. Of the 10 confirmed cases, only four showed a significant change in IgG responses (P < 0.05) between three-weeks PD and follow-up serum specimens by ELISA. Crew members considered confirmed cases consumed significantly more water (P < or = 0.005) aboard the cutter than noncases. Crew members considered EITB positive consumed more water (P < or = 0.04) than crew members considered EITB negative while there was no significant difference in water consumption (P > or

  11. A first survey on the biochemical composition of egg yolk and lysozyme-like activity of egg envelopment in the cuttlefish Sepia officinalis from the Northern Adriatic Sea (Italy).

    PubMed

    Matozzo, Valerio; Conenna, Irene; Riedl, Verena Maria; Marin, Maria Gabriella; Marčeta, Tihana; Mazzoldi, Carlotta

    2015-08-01

    The cuttlefish Sepia officinalis is an important fishery resource in the Northern Adriatic Sea (Italy). During reproduction, fertilised eggs are released by adult females in coastal waters and embryo development can take over two months. During this period, embryos rely on nutrients and other substances, such as immune factors, provided by the female in egg yolk. In cephalopods in general, and specifically in the common cuttlefish, little information is available on yolk biochemical composition and substances included in egg envelopment. In the present study, the main biochemical components of egg yolk and the presence of antimicrobial substances in egg envelopment of S. officinalis were determined for the first time. Statistically significant differences in total egg weight and egg yolk weight were observed among batches from different females. Egg and yolk weights were positively correlated, with yolk representing the 13% (±5%) of the total egg weight. Total proteins were the main biochemical component (46%) of egg yolk, followed by total carbohydrates plus glycogen (39%) and lipids (15%). Statistically significant differences among batches were recorded in egg yolk total protein amounts, lipids, carbohydrates and glycogen, but no correlations were found between egg yolk weight and the biochemical components. The Petri dish and the quantitative spectrophotometric assays revealed the presence of lysozyme-like activity in egg gelatinous envelopment.

  12. Long-term monthly statistics of the mid-latitude ionospheric E-layer peak electron density in the Northern geographic hemisphere during geomagnetically quiet and steadily low solar activity conditions

    NASA Astrophysics Data System (ADS)

    Pavlov, Anatoli; Pavlova, Nadezhda

    2016-07-01

    Long-term hourly values of the ionospheric E-layer peak electron density, NmE, measured during the time period of 1957-2014 by 4 mid-latitude ionosondes (Wallops Island, Boulder, de l'Ebre, and Rome) in the Northern geographic hemisphere were processed to select periods of geomagnetically quiet and low solar activity conditions to calculate several descriptive statistics of NmE close to noon for each month in a year, including the mathematical expectation of NmE, the standard deviations of NmE from the mathematically expected NmE, and the NmE variation coefficient. The month-to-month variability of these descriptors allowed us to identify months of a year when they reach their extremes (maxima, minima). We found that the most probable NmE cannot be considered as the best statistical parameter among the most probable NmE and the mathematically expected NmE in statistical studies of month-to-month variations of NmE. Depending on a choice of an ionosonde and a month, the calculated NmE variation coefficient changes from 5 to 12 %.

  13. Impacts of human activity and climate on the hydrology and ecosystem services of forests in the Guianas on the northern rim of Amazonia

    NASA Astrophysics Data System (ADS)

    Parkin, G.; Wagner, T.; Bovolo, I.; Pereira, R.

    2010-12-01

    Tropical deforestation is one of the main areas of international concern in relation to increases in atmospheric carbon dioxide and feedbacks between land cover and climate. The Guiana Shield region, including the Guianas (Guyana, Surinam, and French Guiana), contains one of the largest remaining continuous areas of tropical forest on the Earth, and is therefore of international importance for climate stability. Its importance to the regional climate of South America is also potentially significant, as a possible conduit for vapour transport between the Caribbean and the continental interior. However, recent studies have confirmed the paucity of reliable hydrometeorological data in the forested interior in the Guianas that could help to improve understanding of these macro-scale links as well as the more local impacts of forest activities. A new hydrological monitoring programme has been established at the Iwokrama International Centre for Rainforest Conservation and Development in the interior of Guyana to provide evidence related to the impacts of forest activities and climate on ecosystem services provided by the forest. These services include water, carbon and nutrient cycling that are essential for maintaining biodiversity and other indicators necessary for long-term sustainability of the forest environment. The Iwokrama Centre aims to test the concept of a truly sustainable forest where conservation, environmental balance and economic use are mutually reinforcing. Iwokrama is uniquely placed for this research, in that it has integral involvement of the indigenous forest communities, and is geographically situated at a transition between mature intact forest and a savannah region that is likely to be sensitive to climate change, and on a larger scale at a transition between distinctive climate types in the coastal region of the Guianas influenced by the Equatorial Trough and the Amazonian interior which experiences a more continental type climate. The

  14. 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.

  15. Occurrence and potential activity of denitrifiers and methanogens in groundwater at 140 m depth in Pliocene diatomaceous mudstone of northern Japan.

    PubMed

    Katsuyama, Chie; Nashimoto, Hiroaki; Nagaosa, Kazuyo; Ishibashi, Tomotaka; Furuta, Kazuki; Kinoshita, Takeshi; Yoshikawa, Hideki; Aoki, Kazuhiro; Asano, Takahiro; Sasaki, Yoshito; Sohrin, Rumi; Komatsu, Daisuke D; Tsunogai, Urumu; Kimura, Hiroyuki; Suwa, Yuichi; Kato, Kenji

    2013-12-01

    Anaerobic microbial activity has a major influence on the subsurface environment. We investigated the denitrification and methanogenesis in anoxic groundwater at a depth of 140 m in two boreholes drilled in a sedimentary geological setting, where the redox potential fluctuated. The average maximum potential denitrification rates, measured under anaerobic conditions in the two boreholes using an (15) N tracer, were 0.060 and 0.085 nmol (30) N2  mL(-1)  h(-1) . The deduced NirS amino acid sequences obtained from in situ samples were similar to those of isolates belonging to the α-, β-, and γ-Proteobacteria, and the Firmicutes (72-100% similarity). Based on the nirS gene, the same operational taxonomic unit dominated incubated samples from each borehole. Methanogenesis candidates were detected by 16S rRNA gene analysis, but no sequence was detected using primers for the functional methanogenesis gene mcrA. Although the stable isotope signatures suggested that some of the dissolved methane was of biogenic origin, no potential for methane production was evident during the incubations. The groundwater at 140 m depth did not contain oxygen, had an Eh ranging from -144 to 6.8 mV, and was found to be a potential field for denitrification.

  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. Structural Analysis of Recent and Active Faults and Regional State of Stress in the Epicentral Area of the 1978 Thessaloniki Earthquakes (northern Greece)

    NASA Astrophysics Data System (ADS)

    Mercier, Jacques-Louis; Carey-Gailhardis, Evelyne; Mouyaris, Nicolas; Simeakis, Konstantinos; Roundoyannis, Theodora; Anghelidhis, Christos

    1983-12-01

    During spring and summer 1978, Macedonia (Greece) suffered damaging seismic activity. During the two main shocks which occurred on May 23 (Mb = 5.7) and June 20 (Mb = 6.1), seismic open cracks and faults were formed in the epicentral area, 25 km NNE from Thessaloniki. They have been mapped and their kinematics analyzed. These studies have shown that they result from the reactivation of existing faults of Pleistocene, Recent, and historical age and that the crust is stretching along a N-S to NNE-SSW direction in that part of the North Aegean. Principal stress directions have been computed from the slip vectors measured on recent neotectonic faults and seismic cracks and faults, this computation utilizing a simple mechanical model (homogeneous deformation through the addition of small and independent displacements of rigid blocks in a highly fractured body). A striking feature of this analysis is that the tensional directions σ3 deduced from seismic faulting and from recent neotectonic faulting are nearly the same. They are compatible with the regional tension axis deduced from focal mechanisms of North Aegean and with the focal mechanisms of the 1978 Thessaloniki main shocks. They are also compatible with in situ stress measurements in this region. Studies of superficial seismic faulting strongly support previous assumptions proposed from aftershock distribution: the 1978 Thessaloniki earthquakes resulted from reactivation of several different faults situated within the same NE dipping fault zone of Upper Eocene-Lower Oligocene age (and older) separating the Serbo-Macedonian Massif from the Peonias (East Vardar) zone.

  18. The Northern Ireland Framework for Peace: Terrorism and its Aftermath

    DTIC Science & Technology

    2012-04-10

    still a risk of Republican destabilization and inflammation of the dissident cause. 22 This was an important factor. The church, the media , and the...use of their criminal enterprises in the day, with involvement in both smuggling and the illegal narcotics market , and their terrorist activity in...Irish taxpayer is effectively setting the level of the Northern Ireland property market .76 The assumption is that all of this is manageable. Northern

  19. Report on Ontario's Northern Schools.

    ERIC Educational Resources Information Center

    2003

    Ontario's funding formula fails to recognize the unique needs of northern school boards, which cover immense geographic areas, have many small schools, and enroll a high proportion of Aboriginal students. This report examines school size, enrollment, and staffing in northern Ontario schools, drawing on 2002-03 tracking reports of provincial…

  20. Active high-resolution seismic tomography of compressional wave velocity and attenuation structure at Medicine Lake Volcano, Northern California Cascade Range

    NASA Astrophysics Data System (ADS)

    Evans, John R.; Zucca, John J.

    1988-12-01

    We determine compressional wave velocity and attenuation structures for the upper crust beneath Medicine Lake volcano in northeast California using a high-resolution active source seismic tomography method. Medicine Lake volcano is a basalt through rhyolite shield volcano of the Cascade Range, lying east of the range axis. The Pg wave from eight explosive sources which has traveled upward through the target volume to a dense array of 140 seismographs provides 1- to 2-km resolution in the upper 5 to 7 km of the crust beneath the volcano. The experiment tests the hypothesis that Cascade Range volcanoes of this type are underlain only by small silicic magma chambers. We image a low-velocity low-Q region not larger than a few tens of cubic kilometers in volume beneath the summit caldera, supporting the hypothesis. A shallower high-velocity high-density feature, previously known to be present, is imaged for the first time in full plan view; it is east-west elongate, paralleling a topographic lineament between Medicine Lake volcano and Mount Shasta. This lineament is interpreted to be the result of an old crustal weakness now affecting the emplacement of magma, both on direct ascent from the lower crust and mantle and in migration from the shallow silicic chamber to summit vents. Differences between this high-velocity feature and the equivalent feature at Newbeny volcano, a volcano in central Oregon resembling Medicine Lake volcano, may partly explain the scarcity of surface hydrothermal features at Medicine Lake volcano. A major low-velocity low-Q feature beneath the southeast flank of the volcano, in an area with no Holocene vents, is interpreted as tephra, flows, and sediments from the volcano deeply ponded on the downthrown side of the Gillem fault, a normal fault mapped at the surface north of the volcano. A high-Q normal-velocity feature beneath the north rim of the summit caldera may be a small, possibly hot, subsolidus intrusion. A high-velocity low-Q region

  1. Early Pleistocene to Holocene glacial activity along the southern Alaska continental shelf inferred from the sedimentary record in the northern Gulf of Alaska - preliminary results

    NASA Astrophysics Data System (ADS)

    Forwick, M.; Cowan, E. A.; Bahlburg, H.; Childress, L. B.; Jaeger, J. M.; Moy, C. M.; Müller, J.; Ribeiro, F.; Ridgway, K. D.; Gulick, S. P.; Worthington, L. L.; Reece, R.

    2013-12-01

    abrupt occurrence of silt laminae and sand beds at the transition from Unit II to Unit I is suggested to reflect a major reorganization of the sediment delivery system to Site U1418. We suggest that these deposits reflect increased turbidity-flow activity related to the first expansion of grounded ice to the shelf break and/or the initiation of ice streams during the Late Pleistocene, transporting large amounts of sediment across the shelf. Slope failures most probably resulted from oversteepening and/or seismicity related to either glacio-isostatic adjustments or tectonic processes. Occasionally occurring different-colored silt laminae indicate that sediments probably originated from multiple sources.

  2. Northern Sinus Meridiani Stereo

    NASA Technical Reports Server (NTRS)

    2003-01-01

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

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

  3. Glyphosate in northern ecosystems.

    PubMed

    Helander, Marjo; Saloniemi, Irma; Saikkonen, Kari

    2012-10-01

    Glyphosate is the main nonselective, systemic herbicide used against a wide range of weeds. Its worldwide use has expanded because of extensive use of certain agricultural practices such as no-till cropping, and widespread application of glyphosate-resistant genetically modified crops. Glyphosate has a reputation of being nontoxic to animals and rapidly inactivated in soils. However, recent evidence has cast doubts on its safety. Glyphosate may be retained and transported in soils, and there may be cascading effects on nontarget organisms. These processes may be especially detrimental in northern ecosystems because they are characterized by long biologically inactive winters and short growing seasons. In this opinion article, we discuss the potential ecological, environmental and agricultural risks of intensive glyphosate use in boreal regions.

  4. Large-scale thrusting along the northern margin of the Tibetan Plateau and the southwest Tarim basin: 230 km long active Hotian thrust sheet

    NASA Astrophysics Data System (ADS)

    Wang, Xin; Suppe, John; Liang, Hang; He, Dengfa

    2014-05-01

    We present the geometry, kinematics and mechanics of large-scale thrusting in the West Kunlun Shan and the southwest Tarim Basin, which is associated with the northward motion of Tibet. The great frontal structure is the ~230km long intact bedding parallel Hotian thrust sheet composed of strata of the Tarim Basin lying above a regional gypsum horizon at the base of the Cenozoic sequence. The toe of the Hotian thrust sheet steps steeply to the surface two thirds of the way across the basin forming the Selibuya-Mazartag hills in the sand desert. The Hotan thrust constitutes one of the longest active intact thrust sheets in the world, showing little internal deformation, however at its back it steps down to a Cambrian detachment at the base of the Paleozoic below a belt of complex high-amplitude anticlines near the front of the West Kunlun Shan, which display break-forward imbricate and wedge structure. More interior, steep reverse faults such as the Tieklik thrust bring older strata to the surface, including Paleozoic basement. The Cambrian detachment also extends northward under the Tarim basin with minor hanging-wall deformation that warps the Hotian Thrust sheet locally, causing the development of growth strata in the Hotian thrust sheet that providesa quantitative record of its motion over these warps. Seismic profiles in the southwest Tarim basin reveal widespread growth strata that record much of the structural history beginning in the early Pliocene Atushi Formation. Ages of seismic reflectors are calibrated to a surface magnetostratigraphic sequence(from Zheng et al., 2000)and traced throughout the seismic grid. The bottom of the growth strata is dated at 3.6 Ma indicating a Pliocene and younger age of thrusting and folding in the southwest Tarim Basin. Structural restoration suggests minimum shortening greater than 35km. The Tieklik thrust consumed at least 10 km in early Pliocene. The fold-and-thrust belts of the southwest Tarim basin shortened >25km in

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

    PubMed

    Miller, Liane

    2016-01-01

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

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

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

  7. Wildfires threaten mercury stocks in northern soils

    USGS Publications Warehouse

    Turetsky, M.R.; Harden, J.W.; Friedli, H.R.; Flannigan, M.; Payne, N.; Crock, J.; Radke, L.

    2006-01-01

    With climate change rapidly affecting northern forests and wetlands, mercury reserves once protected in cold, wet soils are being exposed to burning, likely triggering large releases of mercury to the atmosphere. We quantify organic soil mercury stocks and burn areas across western, boreal Canada for use in fire emission models that explore controls of burn area, consumption severity, and fuel loading on atmospheric mercury emissions. Though renowned as hotspots for the accumulation of mercury and its transformation to the toxic methylmercury, boreal wetlands might soon transition to hotspots for atmospheric mercury emissions. Estimates of circumboreal mercury emissions from this study are 15-fold greater than estimates that do not account for mercury stored in peat soils. Ongoing and projected increases in boreal wildfire activity due to climate change will increase atmospheric mercury emissions, contributing to the anthropogenic alteration of the global mercury cycle and exacerbating mercury toxicities for northern food chains. Copyright 2006 by the American Geophysical Union.

  8. 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

  9. Northern California Supplementary Education Center. Evaluation Report [1970].

    ERIC Educational Resources Information Center

    Chico State Coll., CA. Northern California PACE Center.

    In this evaluation of the Title III (Elementary and Secondary Education Act) Northern California PACE Center program, the center's 1969-70 activities in relation to stated objectives were examined through review of objectives and activities in terms of success in meeting performance requirements. The following objectives were deemed to have met or…

  10. Identification of a novel gene expressed in activated natural killer cells and T cells

    SciTech Connect

    Dahl, C.A.; Schall, R.P.; He, H.; Cairns, J.S. )

    1992-01-15

    The authors have isolated a cDNA clone from a human activated NK cell-derived cDNA library that identifies a transcript [NK4] that is selectively expressed in lymphocytes. The expression of this transcript is increased after activation of T cells by mitogens or activation of NK cells by IL-2 (lymphokine-activated killer cells). The transcript levels demonstrated by Northern blot analysis increase by 12 h after activation, remain high for at least 48 h, and require protein synthesis for expression. Southern blot analysis of B lymphoblastoid lines derived from 18 unrelated individuals reveal variable banding patterns suggestive of polymorphism within the NK4 gene. No homology was found between the sequence of the coding region of this transcript and any sequences in the GenBank data base. Sequence homology to the U1 small nuclear RNA was found within the 3[prime] untranslated region immediately upstream of the site of polyadenylation, suggesting a possible role for U1 in the polyadenylation process. Sequence analysis indicates the transcript would encode a protein having a mass of 27 kDa. The presence of a signal sequence and lack of a transmembrane region suggests that the protein is secreted. In addition, the protein contains an RGD sequence that may be involved in cellular adhesion. This transcript appears to encode a novel product common to the activation pathways of both NK cells and T cells. 50 refs., 8 figs.

  11. Wildfires in Northern Siberian Larch Dominated Communities

    NASA Technical Reports Server (NTRS)

    Khaurk, Viacheslav I.; Ranson, Kenneth J.; Dvinskaya, Maria L.; Im, Sergey T.

    2011-01-01

    The fire history of the northern larch forests within the permafrost zone in a portion of northern Siberia (approx 66 deg N, 100 deg E) was studied. Since there is little to no human activities in this area fires within the study area were mostly caused by lightning. Fire return intervals (FRI) were estimated based on burn marks on tree stems and dates of tree natality. FRI values varied from 130 yr to 350 yr with 200 +/- 50 yr mean. In southerly larch dominated communities FRI was found to be shorter (77 +/- 20 yr at approx 61 deg. N, and 82 +/- 7 at 64 deg N), and longer at the northern boundary (approx 71 deg) of larch stands (320 +/- 50 yr). During the Little Ice Age period in the 16th to 18th centuries FRI was approximately twice as long as recorded in this study. Fire caused changes in the soil including increases in soil drainage and permafrost thawing depth and a radial growth increase of about 2 times (with more than 6 times observed). This effect may simulate the predicted warming impact on the larch growth in the permafrost zone.

  12. MISR Views Northern Australia

    NASA Technical Reports Server (NTRS)

    2000-01-01

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

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

  13. Tornadoes Strike Northern Wisconsin

    NASA Technical Reports Server (NTRS)

    2007-01-01

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

  14. Isolated Northern Dunes

    NASA Technical Reports Server (NTRS)

    2005-01-01

    [figure removed for brevity, see original site]

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

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

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

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

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

  15. Northern Sand Sea

    NASA Technical Reports Server (NTRS)

    2005-01-01

    [figure removed for brevity, see original site]

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

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

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

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

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

  16. Dual infection of chickens with pox and infectious laryngotracheitis (ILT) confirmed with specific pox and ILT DNA dot-blot hybridization assays.

    PubMed

    Fatunmbi, O O; Reed, W M; Schwartz, D L; Tripathy, D N

    1995-01-01

    Dual infection with fowl pox (FP) and infectious laryngotracheitis (ILT) was diagnosed as the cause of acute mortality in a flock of three age groups of Hy-Line leghorn layers. The affected chickens had not been previously vaccinated against either FP or ILT. The diagnosis was confirmed by virus isolation, histopathology, and the use of specific pox and ILT genomic DNA probes in a dot-blot hybridization assay. FP and ILT vaccinations were recommended to control mortality. The use of FP- and ILT-specific DNA dot-blot hybridization may be used as a routine diagnostic tool to differentiate between the two diseases, especially in atypical cases of either infection or to confirm the existence of the two diseases as a mixed infection in a flock of chickens.

  17. 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

  18. Breeding colonies of least terns (Sternula antillarum) in northern Sonora, Mexico, 2006-2008

    USGS Publications Warehouse

    Rosemartin, Alyssa; van Riper, Charles

    2012-01-01

    We document distribution of breeding least terns (Sternula antillarum) in northern Sonora, Mexico, 2006-2008. We report breeding activity at six sites with active colonies, including three previously undocumented colonies.

  19. 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

  20. Stimulation of transforming activity of DJ-1 by Abstrakt, a DJ-1-binding protein.

    PubMed

    Sekito, Aya; Taira, Takahiro; Niki, Takeshi; Iguchi-Ariga, Sanae M M; Ariga, Hiroyoshi

    2005-03-01

    DJ-1 was identified by us as a novel oncogene in cooperation with activated ras. Although over-expression of DJ-1 has been reported in several cancer cells, including cells in breast cancer, lung cancer and prostate cancer, the precise mechanism underlying transformation has not been clarified. In this study, we screened proteins by a yeast two-hybrid method and identified Abstrakt as a DJ-1-binding protein. Abstrakt is an RNA helicase, but it has not yet been characterized. Northern blot analysis showed that human Abstrakt was expressed ubiquitously in all tissues. Abstrakt was then found to bind to and to be colocalized in the nucleus with DJ-1 in human cells. Furthermore, Abstrakt was found to stimulate transforming activity of DJ-1 in rat 3Y1 cells transfected with DJ-1 with activated ras. These findings suggest that Abstrakt is a positive regulator for DJ-1.

  1. Estimating the frequency of volcanic ash clouds over northern Europe

    NASA Astrophysics Data System (ADS)

    Watson, E. J.; Swindles, G. T.; Savov, I. P.; Lawson, I. T.; Connor, C. B.; Wilson, J. A.

    2017-02-01

    Fine ash produced during explosive volcanic eruptions can be dispersed over a vast area, where it poses a threat to aviation, human health and infrastructure. Here, we focus on northern Europe, which lies in the principal transport direction for volcanic ash from Iceland, one of the most active volcanic regions in the world. We interrogate existing and newly produced geological and written records of past ash fallout over northern Europe in the last 1000 years and estimate the mean return (repose) interval of a volcanic ash cloud over the region to be 44 ± 7 years. We compare tephra records from mainland northern Europe, Great Britain, Ireland and the Faroe Islands, with records of proximal Icelandic volcanism and suggest that an Icelandic eruption with a Volcanic Explosivity Index rating (VEI) ≥ 4 and a silicic magma composition presents the greatest risk of producing volcanic ash that can reach northern Europe. None of the ash clouds in the European record which have a known source eruption are linked to a source eruption with VEI < 4. Our results suggest that ash clouds are more common over northern Europe than previously proposed and indicate the continued threat of ash deposition across northern Europe from eruptions of both Icelandic and North American volcanoes.

  2. From the Northern Eurasia Earth Science Partnership Initiative (NEESPI) towards the Northern Eurasia Future Initiative (NEFI)

    NASA Astrophysics Data System (ADS)

    Groisman, Pavel; Gutman, Garik; Gulev, Sergey; Maksyutov, Shamil

    2015-04-01

    Northern Eurasia Earth Science Partnership Initiative (NEESPI, http://neespi.org) was launched in 2004. With its multidisciplinary focus, the internationally funded NEESPI (more than 170 individual international projects during the past decade) has challenged participants to research climate-ecosystem interactions, societal impacts from extreme events in Northern Eurasia, and the feedbacks of these interactions and impacts to the global Earth system. NEESPI was endorsed by major ESSP Programs and Projects and the cornerstone support for the NEESPI studies was provided by the NASA Land Cover and Land Use Change Program and the Russian Academy of Sciences. Now it is the time to synthesis the main achievements of the NEESPI researchers and to re-assess its Science Questions and Objectives of the regional research within the new Future Earth Program paradigm with the focus on interdisciplinary solution-oriented approach that will allow effective policy-making in environment management and control. At the sequence of Workshops (the last of them will be in Prague one week prior to this Assembly) we formulated a major Science Question of the new Northern Eurasia Future Initiative (NEFI): "What will the changes in the regional ecosystems dynamics and interactions mean for the societal well-being, activities, health, and strategic planning in Northern Eurasia?" The major NEFI challenge will be the services aimed on providing in Northern Eurasia a sustainable societal development in changing climate, ecosystems, and societies. At this presentation we shall brief the audience about the main results of the NEESPI researchers, and lay down the plans for the future NEFI studies. At the side event of the Meeting, we shall initiate preparation of the book which will synthesize major NEESPI achievements.

  3. Family Medicine/Northern Medical Services Involvement in Northern Saskatchewan

    PubMed Central

    Irvine, James

    1988-01-01

    To address the problems of recruitment and retention of family physicians in various remote locations in northern Saskatchewan, the University of Saskatchewan became involved through Northern Medical Services, a division of the Department of Family Medicine. The University's involvement consists of the provision of resident family-physician services, visiting consultant services, family-practise resident training, research, and a consulting role of the Medical Health Officer. This paper reviews the context in which this program was created, its role in health care in the area, and its involvement with the communities in health promotion and research. PMID:21253032

  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. 77 FR 50526 - Proposed Safe Harbor Agreement for the Northern Spotted Owl, Skamania, Klickitat, and Yakima...

    Federal Register 2010, 2011, 2012, 2013, 2014

    2012-08-21

    ... Fish and Wildlife Service Proposed Safe Harbor Agreement for the Northern Spotted Owl, Skamania... take of the threatened northern spotted owl (Strix occidentalis caurina) at a level that enables the... would also authorize incidental take of the spotted owl as a result of management activities during...

  6. Discovery of novel hematopoietic cell adhesion molecules from human bone marrow stromal cell membrane protein extracts by a new cell-blotting technique.

    PubMed

    Seshi, B

    1994-05-01

    In an attempt to define the role of cell adhesion molecules (CAMs) within the bone marrow (BM) microenvironment in normal hematopoiesis and in leukemia development, a novel cell-blotting technique that involved cell adhesion to protein bands after separation by lithium dodecyl sulfate-polyacrylamide gel electrophoresis (LDS-PAGE) and blotting onto polyvinylidene difluoride (PVDF) membrane has been developed. Human BM stromal cell membrane fractions have been prepared from Dexter-type cultures after cell lysis by sonification and differential centrifugations of the sonification contents. The 20,000 g pellets representing membrane fractions have been solubilized by 2% Triton X-100, 0.575% LDS, and 8 mol/L urea in sequential order. The protein extracts are fractionated by LDS-PAGE and screened for CAMs by the new cell-blotting technique. This led to identification of nine protein bands in lanes containing LDS extracts showing adhesion of KG1a (CD34+ progenitor myeloid) cells. Evidence that the BM proteins exhibiting KG1a cell adhesion are novel CAMs is based on the observations that these proteins, in comparison with known CAMs, specifically VCAM-1, CD54, and CD44, show (1) contrasting detergent-solubility properties, (2) different temperature requirement for mediating cell adhesion function, and (3) markedly distinct electrophoretic mobilities. The various cell types tested, notably KG1a, NALM-6, WIL-2, Ramos, HS-Sultan, K562, JY B lymphoblastoid cells, and T lymphoblasts, showed distinctive patterns of binding to different subsets of BM CAMs. These results demonstrate a new approach to studies of molecular mechanisms that may determine specificity of hematopoietic cellular localization within BM microenvironment and may play an important role in controlling hematopoiesis.

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

    PubMed Central

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

    2000-01-01

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

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

    PubMed Central

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

    2015-01-01

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

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

    NASA Technical Reports Server (NTRS)

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

    2012-01-01

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

  10. Titan's stratospheric condensibles at high northern latitudes during northern winter

    NASA Astrophysics Data System (ADS)

    Anderson, C.; Samuelson, R.; Achterberg, R.

    2012-04-01

    The Infrared Interferometer Spectrometer (IRIS) instrument on board Voyager 1 caught the first glimpse of an unidentified particulate feature in Titan’s stratosphere that spectrally peaks at 221 cm-1. Until recently, this feature that we have termed ‘the haystack,’ has been seen persistently at high northern latitudes with the Composite Infrared Spectrometer (CIRS) instrument onboard Cassini. The strength of the haystack emission feature diminishes rapidly with season, becoming drastically reduced at high northern latitudes, as Titan transitions from northern winter into spring. In contrast to IRIS whose shortest wavenumber was 200 cm-1, CIRS extends down to 10 cm-1, thus revealing an entirely unexplored spectral region in which nitrile ices have numerous broad lattice vibration features. Unlike the haystack, which is only found at high northern latitudes during northern winter/early northern spring, this geometrically thin nitrile cloud pervades Titan’s lower stratosphere, spectrally peaking at 160 cm-1, and is almost global in extent spanning latitudes 85°N to 60°S. The inference of nitrile ices are consistent with the highly restricted altitude ranges over which these features are observed, and appear to be dominated by a mixture of HCN and HC3N. The narrow range in altitude over which the nitrile ices extend is unlike the haystack, whose vertical distribution is significantly broader, spanning roughly 70 km in altitude in Titan’s lower stratosphere. The nitrile clouds that CIRS observes are located in a dynamically stable region of Titan’s atmosphere, whereas CH4 clouds, which ordinarily form in the troposphere, form in a more dynamically unstable region, where convective cloud systems tend to occur. In the unusual situation where Titan’s tropopause cools significantly from the HASI 70.5K temperature minimum, CH4 should condense in Titan’s lower stratosphere, just like the aforementioned nitrile clouds, although in significantly larger

  11. Opsin gene repertoires in northern archaic hominids.

    PubMed

    Taylor, John S; Reimchen, Thomas E

    2016-08-01

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

  12. Grazing effects on carbon fluxes in a northern China grassland

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Grazing is a widespread use of grasslands in northern China, but if stocking rate exceeds grassland carrying capacity, degradation and desertification can occur. As a result, grazing management is critical and can play a significant role in driving C sink and source activity in grassland ecosystems...

  13. TORC1 activity is partially reduced under nitrogen starvation conditions in sake yeast Kyokai no. 7, Saccharomyces cerevisiae.

    PubMed

    Nakazawa, Nobushige; Sato, Aya; Hosaka, Masahiro

    2016-03-01

    Industrial yeasts are generally unable to sporulate but treatment with the immunosuppressive drug rapamycin restores this ability in a sake yeast strain Kyokai no. 7 (K7), Saccharomyces cerevisiae. This finding suggests that TORC1 is active under sporulation conditions. Here, using a reporter gene assay, Northern and Western blots, we tried to gain insight into how TORC1 function under nitrogen starvation conditions in K7 cells. Similarly to a laboratory strain, RPS26A transcription was repressed and Npr1 was dephosphorylated in K7 cells, indicative of the expected loss of TORC1 function under nitrogen starvation. The expression of nitrogen catabolite repression-sensitive genes, however, was not induced, the level of Cln3 remained constant, and autophagy was more slowly induced than in a laboratory strain, all suggestive of active TORC1. We conclude that TORC1 activity is partially reduced under nitrogen starvation conditions in K7 cells.

  14. Primary Science in Northern Ireland

    ERIC Educational Resources Information Center

    McAllister, Peter

    2005-01-01

    Since 1990 the science curriculum in Northern Ireland has gone through three major changes. In the beginning, fifteen attainment targets were introduced to an unsuspecting and largely unprepared teaching population: these were eventually reduced to five in 1993 and then to the present two in 1996. Unlike in England, technology has never stood as…

  15. "Subtractive" Bilingualism in Northern Belize.

    ERIC Educational Resources Information Center

    Rubinstein, Robert A.

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

  16. Motivating Learners in Northern Communities.

    ERIC Educational Resources Information Center

    Swanson, Sharon

    2003-01-01

    A teacher at a northern Ontario adult Native literacy program describes how she cultivates student motivation. The teacher-student relationship is the most influential factor for motivating students. By focusing on cultural awareness, cultural teaching practices, and a sense of community, teachers can help students be successful. Program…

  17. [RELATIONSHIPS IN THE "NORTHERN FULMAR (FULMAR US GLACIALIS)--TETRABOTHRIUS MINOR (CESTODA: TETRABOTHRIIDAE)" SYSTEM: PHYSIOLOGICAL ASPECTS].

    PubMed

    Kuklina, M M

    2015-01-01

    Relationships between the northern fulmar (Fulmarus glacialis L., 1761) and cestodes Tetrabothrius minor Loennberg, 1893 (Cestoda: Tetrabothriidae) were studied. The results of calculation of the number of tapeworms in different parts (proximal, medial and distal) of bird intestines are represented. Parameters of protein metabolism in the northern fulmar and cestodes T. minor were investigated. Activity of proteases in different parts of northern fulmar intestine and in the strobila of T. minor was determined. Digestion processes occurring on digestive-absorptive surfaces of the intestine of the northern fulmar and of cestodes were studied. Biochemical indices of blood plasma of the northern fulmar were analyzed in relation to the intensity of invasion and the stage parasite maturation. The highest indices of the invasion of the northern fulmar with T. minor were recorded in the proximal part if the intestine. It was shown that the preferred localization of tapeworms in the proximal department of the intestine was determined by abundance of food and high activity of digestive enzymes in this place. Active hydrolysis of proteins in the intestine of the northern fulmar and on tegument surfaces of T. minor occurred mainly during the process of cavernous digestion. To a greater extent, the physiological state of the northern fulmar depended on the intensity of invasion and on the maturation stage of tapeworms.

  18. Evaporite karst of northern lower Michigan

    USGS Publications Warehouse

    Black, T.J.

    1997-01-01

    Michigan has three main zones of evaporite karst: collapse breccia in Late Silurian deposits of the Mackinac Straits region; breccia, collapse sinks, and mega-block collapse in Middle Devonian deposits of Northern Lower Michigan, which overlaps the preceding area; and areas of soil swallows in sinks of Mississippian deposits between Turner and Alabaster in Arenac and Iosco counties, and near Grand Rapids in Kent County. The author has focused his study on evaporite karst of the Middle Devonian deposits. The Middle Devonian depos its are the Detroit River Group: a series consisting of limestone, dolomite, shale, salt, gypsum, and anhydrite. The group occurs from subcrop, near the surface, to nearly 1400 feet deep from the northern tip of the Southern Peninsula to the south edge of the "solution front" Glacial drift is from zero to 350 feet thick. Oil and gas exploration has encountered some significant lost-circulation zones throughout the area. Drilling without fluid returns, casing-seal failures, and lost holes are strong risks in some parts of the region. Lost fluid returns near the top of the group in nearby areas indicate some karst development shortly after deposition. Large and irregular lost-circulation zones, linear and patch trends of large sink holes, and 0.25 mile wide blocks of down-dropped land in the northern Lower Peninsula of Michigan were caused by surface- and ground-water movement along faults into the Detroit River Group. Glaciation has removed some evidence of the karst area at the surface. Sinkhole development, collapse valleys, and swallows developed since retreat of the glacier reveal an active solution front in the Detroit River Group.

  19. Northern Border Pipeline Company NPDES Permit

    EPA Pesticide Factsheets

    Under NPDES permit MT-0030791, the Northern Border Pipeline Company is authorized to discharge from locations along the Northern Border Gas Transmission Pipeline located within the exterior boundaries of the Fort Peck Indian Reservation, Montana.

  20. 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.

  1. 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.

  2. 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.

  3. Polymerase chain reaction and Southern blot-based analysis of the C9orf72 hexanucleotide repeat in different motor neuron diseases.

    PubMed

    Hübers, Annemarie; Marroquin, Nicolai; Schmoll, Birgit; Vielhaber, Stefan; Just, Marlies; Mayer, Benjamin; Högel, Josef; Dorst, Johannes; Mertens, Thomas; Just, Walter; Aulitzky, Anna; Wais, Verena; Ludolph, Albert C; Kubisch, Christian; Weishaupt, Jochen H; Volk, Alexander E