WorldWideScience

Sample records for western blot method

  1. Western blotting.

    Science.gov (United States)

    Kurien, Biji T; Scofield, R Hal

    2006-04-01

    Western blotting (protein blotting or immunoblotting) is a powerful and important procedure for the immunodetection of proteins post-electrophoresis, particularly proteins that are of low abundance. Since the inception of the protocol for protein transfer from an electrophoresed gel to a membrane in 1979, protein blotting has evolved greatly. The scientific community is now confronted with a variety of ways and means to carry out this transfer. This review describes the various procedures that have been used to transfer proteins from a gel to a membrane based on the principles of simple diffusion, vacuum-assisted solvent flow and electrophoretic elution. Finally, a brief description of methods generally used to detect antigens on blots is also described.

  2. [Better performance of Western blotting: quick vs slow protein transfer, blotting membranes and the visualization methods].

    Science.gov (United States)

    Kong, Ling-Quan; Pu, Ying-Hui; Ma, Shi-Kun

    2008-01-01

    To study how the choices of the quick vs slow protein transfer, the blotting membranes and the visualization methods influence the performance of Western blotting. The cellular proteins were abstracted from human breast cell line MDA-MB-231 for analysis with Western blotting using quick (2 h) and slow (overnight) protein transfer, different blotting membranes (nitrocellulose, PVDF and nylon membranes) and different visualization methods (ECL and DAB). In Western blotting with slow and quick protein transfer, the prestained marker presented more distinct bands on nitrocellulose membrane than on the nylon and PVDF membranes, and the latter also showed clear bands on the back of the membrane to very likely cause confusion, which did not occur with nitrocellulose membrane. PVDF membrane allowed slightly clearer visualization of the proteins with DAB method as compared with nitrocellulose and nylon membranes, and on the latter two membranes, quick protein transfer was likely to result in somehow irregular bands in comparison with slow protein transfer. With slow protein transfer and chemiluminescence for visualization, all the 3 membranes showed clear background, while with quick protein transfer, nylon membrane gave rise to obvious background noise but the other two membranes did not. Different membranes should be selected for immunoblotting according to the actual needs of the experiment. Slow transfer of the proteins onto the membranes often has better effect than quick transfer, and enhanced chemiluminescence is superior to DAB for protein visualization and allows highly specific and sensitive analysis of the protein expressions.

  3. Western blotting: an introduction.

    Science.gov (United States)

    Kurien, Biji T; Scofield, R Hal

    2015-01-01

    Western blotting is an important procedure for the immunodetection of proteins, particularly proteins that are of low abundance. This process involves the transfer of protein patterns from gel to microporous membrane. Electrophoretic as well as non-electrophoretic transfer of proteins to membranes was first described in 1979. Protein blotting has evolved greatly since the inception of this protocol, allowing protein transfer to be accomplished in a variety of ways.

  4. Western Blotting using Capillary Electrophoresis

    OpenAIRE

    Anderson, Gwendolyn J.; Cipolla, Cynthia; Kennedy, Robert T.

    2011-01-01

    A microscale Western blotting system based on separating sodium-dodecyl sulfate protein complexes by capillary gel electrophoresis followed by deposition onto a blotting membrane for immunoassay is described. In the system, the separation capillary is grounded through a sheath capillary to a mobile X-Y translation stage which moves a blotting membrane past the capillary outlet for protein deposition. The blotting membrane is moistened with a methanol and buffer mixture to facilitate protein a...

  5. Western blotting using capillary electrophoresis.

    Science.gov (United States)

    Anderson, Gwendolyn J; M Cipolla, Cynthia; Kennedy, Robert T

    2011-02-15

    A microscale Western blotting system based on separating sodium-dodecyl sulfate protein complexes by capillary gel electrophoresis followed by deposition onto a blotting membrane for immunoassay is described. In the system, the separation capillary is grounded through a sheath capillary to a mobile X-Y translation stage which moves a blotting membrane past the capillary outlet for protein deposition. The blotting membrane is moistened with a methanol and buffer mixture to facilitate protein adsorption. Although discrete protein zones could be detected, bands were broadened by ∼1.7-fold by transfer to membrane. A complete Western blot for lysozyme was completed in about one hour with 50 pg mass detection limit from low microgram per milliliter samples. These results demonstrate substantial reduction in time requirements and improvement in mass sensitivity compared to conventional Western blots. Western blotting using capillary electrophoresis shows promise to analyze low volume samples with reduced reagents and time, while retaining the information content of a typical Western blot.

  6. Western blotting using chemiluminescent substrates.

    Science.gov (United States)

    Alegria-Schaffer, Alice

    2014-01-01

    Western blotting is a powerful and commonly used tool to identify and quantify a specific protein in a complex mixture (Towbin et al., 1979). The technique enables indirect detection of protein samples immobilized on a nitrocellulose or polyvinylidene fluoride (PVDF) membrane. Copyright © 2014 Elsevier Inc. All rights reserved.

  7. Multiplexed Western Blotting Using Microchip Electrophoresis.

    Science.gov (United States)

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

    2016-07-05

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

  8. Western Blotting of the Endocannabinoid System.

    Science.gov (United States)

    Wager-Miller, Jim; Mackie, Ken

    2016-01-01

    Measuring expression levels of G protein-coupled receptors (GPCRs) is an important step for understanding the distribution, function, and regulation of these receptors. A common approach for detecting proteins from complex biological systems is Western blotting. In this chapter, we describe a general approach to Western blotting protein components of the endocannabinoid system using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and nitrocellulose membranes, with a focus on detecting type 1 cannabinoid (CB1) receptors. When this technique is carefully used, specifically with validation of the primary antibodies, it can provide quantitative information on protein expression levels. Additional information can also be inferred from Western blotting such as potential posttranslational modifications that can be further evaluated by specific analytical techniques.

  9. The Fastest Western in Town: A Contemporary Twist on the Classic Western Blot Analysis

    OpenAIRE

    Silva, Jillian M.; McMahon, Martin

    2014-01-01

    The Western blot techniques that were originally established in the late 1970s are still actively utilized today. However, this traditional method of Western blotting has several drawbacks that include low quality resolution, spurious bands, decreased sensitivity, and poor protein integrity. Recent advances have drastically improved numerous aspects of the standard Western blot protocol to produce higher qualitative and quantitative data. The Bis-Tris gel system, an alternative to the convent...

  10. Western Blot of Stained Proteins from Dried Polyacrylamide Gels

    Science.gov (United States)

    Gruber, Claudia; Stan-Lotter, Helga

    1996-01-01

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

  11. The Design of a Quantitative Western Blot Experiment

    Directory of Open Access Journals (Sweden)

    Sean C. Taylor

    2014-01-01

    Full Text Available Western blotting is a technique that has been in practice for more than three decades that began as a means of detecting a protein target in a complex sample. Although there have been significant advances in both the imaging and reagent technologies to improve sensitivity, dynamic range of detection, and the applicability of multiplexed target detection, the basic technique has remained essentially unchanged. In the past, western blotting was used simply to detect a specific target protein in a complex mixture, but now journal editors and reviewers are requesting the quantitative interpretation of western blot data in terms of fold changes in protein expression between samples. The calculations are based on the differential densitometry of the associated chemiluminescent and/or fluorescent signals from the blots and this now requires a fundamental shift in the experimental methodology, acquisition, and interpretation of the data. We have recently published an updated approach to produce quantitative densitometric data from western blots (Taylor et al., 2013 and here we summarize the complete western blot workflow with a focus on sample preparation and data analysis for quantitative western blotting.

  12. Multistrip Western blotting to increase quantitative data output

    OpenAIRE

    Kiyatkin, Anatoly; Aksamitiene, Edita

    2009-01-01

    The qualitative and quantitative measurements of protein abundance and modification states are essential in understanding their functions in diverse cellular processes. Typical Western blotting, though sensitive, is prone to produce substantial errors and is not readily adapted to high-throughput technologies. Multistrip Western blotting is a modified immunoblotting procedure based on simultaneous electrophoretic transfer of proteins from multiple strips of polyacrylamide gels to a single mem...

  13. Multistrip western blotting to increase quantitative data output.

    Science.gov (United States)

    Kiyatkin, Anatoly; Aksamitiene, Edita

    2009-01-01

    The qualitative and quantitative measurements of protein abundance and modification states are essential in understanding their functions in diverse cellular processes. Typical western blotting, though sensitive, is prone to produce substantial errors and is not readily adapted to high-throughput technologies. Multistrip western blotting is a modified immunoblotting procedure based on simultaneous electrophoretic transfer of proteins from multiple strips of polyacrylamide gels to a single membrane sheet. In comparison with the conventional technique, Multistrip western blotting increases the data output per single blotting cycle up to tenfold, allows concurrent monitoring of up to nine different proteins from the same loading of the sample, and substantially improves the data accuracy by reducing immunoblotting-derived signal errors. This approach enables statistically reliable comparison of different or repeated sets of data, and therefore is beneficial to apply in biomedical diagnostics, systems biology, and cell signaling research.

  14. Determination of Diagnostic Antigens in Cattle Amphistomiasis Using Western Blotting

    Directory of Open Access Journals (Sweden)

    A Halajian

    2009-05-01

    Full Text Available "nBackground: Mixed infection with amphistomes seems common in native cattle of Iran. The aim of this study was to determine diagnostic antigens in cattle mixed amphistomiasis."nMethods: Specific antigens of Cotylophoron cotylophorum, Gastrothylax crumenifer and Paramphisto­mum cervi (mixed infection, the most common species, were collected from cattle was deter­mined. Adult trematodes were collected from the rumen of naturally infected cattle at meat inspec­tion. After their homogenization and centrifugation, somatic antigens were prepared and ana­lyzed by SDS-PAGE. Specific antigens were determinated by western blot with homologous and heterolo­gous sera. SDS-PAGE of whole worms extract was performed at different concentrations and subse­quent gels staining. Immunoblotting analysis using sera from cattle naturally infected with am­phistomes, Dicrocoelium dendriticum, Fasciola spp. and hydatid cyst was performed."nResults: Electrophorese analysis of somatic antigens revealed the presence of 10 and 21 protein bands at 4 µgr/ml and 8 µgr/ml with molecular weights ranging from 25-120 and 25-150 kDa, respectively. The best result was taken at 8 mg/ml concentration. Although western blot of these proteins demon­strate 5 major antigenic polypeptides ranging from 50 to 100 kDa which were recognized by serum of cat­tle naturally infected with mixed amphistomes.

  15. Ultrasensitive Detection of Proteins on Western Blots with Semiconducting Polymer Dots

    OpenAIRE

    Ye, Fangmao; Smith, Polina B.; Wu, Changfeng; Chiu, Daniel T.

    2013-01-01

    We demonstrate ultrasensitive fluorescence imaging of proteins on Western blots using a bright, compact, and orange-emitting semiconducting polymer dot (CN-PPV). We achieved a detection limit at the single-picogram level in dot blots; with conventional Western blotting, we detected 50 pg of transferrin and trypsin inhibitor after SDS-PAGE and transfer onto a PVDF membrane. Our method does not require any additional equipment or time compared to the conventional procedure with traditional fluo...

  16. Routine Western blot to check autophagic flux : Cautions and recommendations

    NARCIS (Netherlands)

    Gomez-Sanchez, Ruben; Pizarro-Estrella, Elisa; Yakhine-Diop, Sokhna M. S.; Rodriguez-Arribas, Mario; Bravo-San Pedro, Jose M.; Fuentes, Jose M.; Gonzalez-Polo, Rosa A.

    2015-01-01

    At present, the analysis of autophagic flux by Western blotting (WB), which measures two of the most important markers of autophagy, i.e., microtubule-associated protein 1 light chain 3 (LC3) and p62, is widely accepted in the scientific community. In this study, we addressed the possible

  17. Conditions that allow for effective transfer of membrane proteins onto nitrocellulose membrane in Western blots.

    Science.gov (United States)

    Abeyrathne, Priyanka D; Lam, Joseph S

    2007-04-01

    A major hurdle in characterizing bacterial membrane proteins by Western blotting is the ineffectiveness of transferring these proteins from sodium dodecyl sulfate -- polyacrylamide gel electrophoresis (SDS-PAGE) gel onto nitrocellulose membrane, using standard Western blot buffers and electrophoretic conditions. In this study, we compared a number of modified Western blotting buffers and arrived at a composition designated as the SDS-PAGE-Urea Lysis buffer. The use of this buffer and specific conditions allowed the reproducible transfer of highly hydrophobic bacterial membrane proteins with 2-12 transmembrane-spanning segments as well as soluble proteins onto nitrocellulose membranes. This method should be broadly applicable for immunochemical studies of other membrane proteins.

  18. Multi-strip Western blotting to increase quantitative data output

    OpenAIRE

    Aksamitiene, Edita; Hoek, Jan B.; Kholodenko, Boris; Kiyatkin, Anatoly

    2007-01-01

    The qualitative and quantitative measurement of protein abundance and protein modification states are essential in understanding their role in diverse cellular processes. Traditional Western blotting technique, though sensitive, is prone to produce substantial errors and is not readily adapted to high-throughput technologies. We propose a modified immunoblotting procedure, which is based on simultaneous transfer of proteins from multiple gel-strips onto the same membrane, and is compatible wi...

  19. Cy5 total protein normalization in Western blot analysis.

    Science.gov (United States)

    Hagner-McWhirter, Åsa; Laurin, Ylva; Larsson, Anita; Bjerneld, Erik J; Rönn, Ola

    2015-10-01

    Western blotting is a widely used method for analyzing specific target proteins in complex protein samples. Housekeeping proteins are often used for normalization to correct for uneven sample loads, but these require careful validation since expression levels may vary with cell type and treatment. We present a new, more reliable method for normalization using Cy5-prelabeled total protein as a loading control. We used a prelabeling protocol based on Cy5 N-hydroxysuccinimide ester labeling that produces a linear signal response. We obtained a low coefficient of variation (CV) of 7% between the ratio of extracellular signal-regulated kinase (ERK1/2) target to Cy5 total protein control signals over the whole loading range from 2.5 to 20.0μg of Chinese hamster ovary cell lysate protein. Corresponding experiments using actin or tubulin as controls for normalization resulted in CVs of 13 and 18%, respectively. Glyceraldehyde-3-phosphate dehydrogenase did not produce a proportional signal and was not suitable for normalization in these cells. A comparison of ERK1/2 signals from labeled and unlabeled samples showed that Cy5 prelabeling did not affect antibody binding. By using total protein normalization we analyzed PP2A and Smad2/3 levels with high confidence. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. Acid-Urea Gel Electrophoresis and Western Blotting of Histones.

    Science.gov (United States)

    Hazzalin, Catherine A; Mahadevan, Louis C

    2017-01-01

    Acid-urea gel electrophoresis offers significant advantages over SDS-PAGE for analysis of post-translational protein modifications, being capable of resolving proteins of similar size but varying in charge. Hence, it can be used to separate protein variants with small charge-altering differences in primary sequence, and is particularly useful in the analysis of histones whose charge variation arises from post-translational modification, such as phosphorylation or acetylation. On acid-urea gels, histones that carry multiple modifications, each with a characteristic charge, are resolved into distinct bands, the so-called "histone ladder." Thus, the extent and distribution of different modification states of histones can be visualized. Here, we describe the analysis of histone H3 by acid-urea gel electrophoresis and western blotting.

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

    Science.gov (United States)

    Kurien, Biji T; Scofield, R Hal

    2009-01-01

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

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

    Science.gov (United States)

    Kurien, Biji T; Scofield, R Hal

    2015-01-01

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

  3. Fluorescence-based Western blotting for quantitation of protein biomarkers in clinical samples.

    Science.gov (United States)

    Zellner, Maria; Babeluk, Rita; Diestinger, Michael; Pirchegger, Petra; Skeledzic, Senada; Oehler, Rudolf

    2008-09-01

    Since most high throughput techniques used in biomarker discovery are very time and cost intensive, highly specific and quantitative analytical alternative application methods are needed for the routine analysis. Conventional Western blotting allows detection of specific proteins to the level of single isotypes while its quantitative accuracy is rather limited. We report a novel and improved quantitative Western blotting method. The use of fluorescently labelled secondary antibodies strongly extends the dynamic range of the quantitation and improves the correlation with the protein amount (r=0.997). By an additional fluorescent staining of all proteins immediately after their transfer to the blot membrane, it is possible to visualise simultaneously the antibody binding and the total protein profile. This allows for an accurate correction for protein load. Applying this normalisation it could be demonstrated that fluorescence-based Western blotting is able to reproduce a quantitative analysis of two specific proteins in blood platelet samples from 44 subjects with different diseases as initially conducted by 2D-DIGE. These results show that the proposed fluorescence-based Western blotting is an adequate application technique for biomarker quantitation and suggest possibilities of employment that go far beyond.

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

    Science.gov (United States)

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

    2016-01-01

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

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

    Science.gov (United States)

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

    2016-09-01

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

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

    Science.gov (United States)

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

    2012-01-01

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

  7. Western Blotting Using the Invitrogen NuPage Novex Bis Tris MiniGels

    OpenAIRE

    Penna, Aubin; Cahalan, Michael

    2007-01-01

    Western Blotting (or immunoblotting) is a standard laboratory procedure allowing investigators to verify the expression of a protein, determine the relative amount of the protein present in different samples, and analyze the results of co-immunoprecipitation experiments. In this method, a target protein is detected with a specific primary antibody in a given sample of tissue homogenate or extract. Protein separation according to molecular weight is achieved using denaturing SDS-PAGE. After tr...

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

    Science.gov (United States)

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

    2012-01-01

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

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

    Science.gov (United States)

    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…

  10. Banding pattern indicative of echinococcosis in a commercial cysticercosis western blot

    Directory of Open Access Journals (Sweden)

    Tappe D

    2009-09-01

    Full Text Available Abstract Objective A commercial cysticercosis Western blot was evaluated for serological cross-reactivity of sera from patients with alveolar (AE and cystic echinococcosis (CE. Methods A total of 161 sera were examined, including 31 sera from AE-patients, 11 sera from CE-patients, 9 sera from patients with other parasitic diseases and 109 sera from patients with unrelated medical conditions. All AE-and CE-sera were also examined by the echinococcosis Western blot. Results More sera from patients with AE than with CE showed cross-reactivity in the form of ladder-like patterns ("Mikado aspect" and untypical bands at 6-8 kDa (71% and 77.4% versus 27.3% and 45.5%, respectively. In contrast, triplets of bands in the area above 50 kDa and between 24 and 39-42 kDa were more frequent in CE than in AE sera. The fuzzy band at 50-55 kDa typical for cysticercosis was absent in all AE and CE sera. Conclusions Atypical banding patterns in the cysticercosis Western blot should raise the suspicion of a metacestode infection different from Taenia solium, i.e. Echinococcus multilocularis or E. granulosus, especially when the Mikado aspect and an altered 6-8 kDa band is visible in the absence of a fuzzy 50-55 kDa band.

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

    Science.gov (United States)

    García-Solaesa, Virginia; Abad, Sara Ciria

    2016-01-01

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

  12. Phenylglyoxal-Based Visualization of Citrullinated Proteins on Western Blots

    Directory of Open Access Journals (Sweden)

    Sanne M. M. Hensen

    2015-04-01

    Full Text Available Citrullination is the conversion of peptidylarginine to peptidylcitrulline, which is catalyzed by peptidylarginine deiminases. This conversion is involved in different physiological processes and is associated with several diseases, including cancer and rheumatoid arthritis. A common method to detect citrullinated proteins relies on anti-modified citrulline antibodies directed to a specific chemical modification of the citrulline side chain. Here, we describe a versatile, antibody-independent method for the detection of citrullinated proteins on a membrane, based on the selective reaction of phenylglyoxal with the ureido group of citrulline under highly acidic conditions. The method makes use of 4-azidophenylglyoxal, which, after reaction with citrullinated proteins, can be visualized with alkyne-conjugated probes. The sensitivity of this procedure, using an alkyne-biotin probe, appeared to be comparable to the antibody-based detection method and independent of the sequence surrounding the citrulline.

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

    Directory of Open Access Journals (Sweden)

    D.V. Pilonetto

    2009-03-01

    Full Text Available 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.

  14. Multistrip Western blotting: a tool for comparative quantitative analysis of multiple proteins.

    Science.gov (United States)

    Aksamitiene, Edita; Hoek, Jan B; Kiyatkin, Anatoly

    2015-01-01

    The qualitative and quantitative measurements of protein abundance and modification states are essential in understanding their functions in diverse cellular processes. Typical Western blotting, though sensitive, is prone to produce substantial errors and is not readily adapted to high-throughput technologies. Multistrip Western blotting is a modified immunoblotting procedure based on simultaneous electrophoretic transfer of proteins from multiple strips of polyacrylamide gels to a single membrane sheet. In comparison with the conventional technique, Multistrip Western blotting increases data output per single blotting cycle up to tenfold; allows concurrent measurement of up to nine different total and/or posttranslationally modified protein expression obtained from the same loading of the sample; and substantially improves the data accuracy by reducing immunoblotting-derived signal errors. This approach enables statistically reliable comparison of different or repeated sets of data and therefore is advantageous to apply in biomedical diagnostics, systems biology, and cell signaling research.

  15. Checking transfer efficiency and equal loading via qualitative optical way in western blotting.

    Science.gov (United States)

    Gong, Jun-Hua; Gong, Jian-Ping; Zheng, Kai-Wen

    2017-11-01

    The ability to determine that successful transfer and equal loading occur prior to using primary antibodies is important. And total protein staining is commonly used to check transfer efficiency and normalization, which play a crucial role in western blotting. Ponceau S and coomassie blue are commonly used, but there are disadvantages reported in recent years. Therefore, we are interested in finding another method, which is cheap, easy and fast. As we know, protein binding region of PVDF membrane is still hydrophilic when carbinol volatilizes, however, the non-protein binding region of PVDF membrane became hydrophobic again. And this different wettability between non-protein binding region and protein binding region of Polyvinylidene difluoride membrane may be used to check transfer efficiency and equal loading in western blotting. Based on the principle above, we describe an optical approach where an experimenter can observe that the proteins have been transferred to the membrane without any staining within minutes. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. An alternative method for processing northern blots after capillary transfer.

    Science.gov (United States)

    Nilsen, Timothy W

    2015-03-02

    Different laboratories use different methods for the prehybridization, hybridization, and washing steps of the northern blotting procedure. In this protocol, a northern blot is pretreated with Church and Gilbert hybridization buffer to block nonspecific probe-binding sites. The immobilized RNA is then hybridized to a DNA probe specific for the RNA of interest. Finally, the membrane is washed and subjected to autoradiography or phosphorimaging. The solutions and conditions described here may be ideal for those who prefer to use fewer ingredients in their solutions. This protocol is designed to achieve the same goals as other northern blotting approaches. It minimizes background (nonspecific adherence of probe to membrane and nonspecific hybridization) and maximizes specific hybridization to RNAs immobilized on a membrane. © 2015 Cold Spring Harbor Laboratory Press.

  17. Should we ignore western blots when selecting antibodies for other applications?

    DEFF Research Database (Denmark)

    Uhlén, Mathias

    2017-01-01

    .In the Human Protein Atlas (HPA) program, we have validated more than 24,000 in-house-generated antibodies directed to 17,000 human target proteins2. Although there is often a correlation between performance in different applications, we have observed many examples of antibodies that show strong support...... applications and that this influences the epitopes exposed on the target protein, which might have profound consequences for the ability of a given antibody to bind specifically to its target. As an example, proteins that are analyzed by immunohistochemistry (IHC) are normally first cross-linked with formalin.......In conclusion, western blot and protein array analyses can indeed be useful tools when selecting specific antibodies for other applications. The use of these methods is encouraged both for antibody providers and users, and antibodies with signs of cross-reactivity in these applications should be treated...

  18. Improvement of western blot test specificity for detecting equine serum antibodies to Sarcocystis neurona.

    Science.gov (United States)

    Rossano, M G; Mansfield, L S; Kaneene, J B; Murphy, A J; Brown, C M; Schott, H C; Fox, J C

    2000-01-01

    Equine protozoal myeloencephalitis (EPM) is a neurological disease of horses and ponies caused by the apicomplexan protozoan parasite Sarcocystis neurona. The purposes of this study were to develop the most stringent criteria possible for a positive test result, to estimate the sensitivity and specificity of the EPM Western blot antibody test, and to assess the ability of bovine antibodies to Sarcocystis cruzi to act as a blocking agent to minimize false-positive results in the western blot test for S. neurona. Sarcocystis neurona merozoites harvested from equine dermal cell culture were heat denatured, and the proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis in a 12-20% linear gradient gel. Separated proteins were electrophoretically transferred to polyvinylidene fluoride membranes and blocked in 1% bovine serum albumin and 0.5% Tween-Tris-buffered saline. Serum samples from 6 horses with S. neurona infections (confirmed by culture from neural tissue) and 57 horses without infections (horses from the Eastern Hemisphere, where S. neurona does not exist) were tested by Western blot. Horses from both groups had reactivity to the 62-, 30-, 16-, 13-, 11-, 10.5-, and 10-kD bands. Testing was repeated with another step. Blots were treated with bovine S. cruzi antibodies prior to loading the equine samples. After this modification of the Western blot test, positive infection status was significantly associated with reactivity to the 30- and 16-kD bands (Pblot had a sample sensitivity of 100% and sample specificity of 98%. It is concluded that the specificity of the Western blot test is improved by blocking proteins not specific to S. neurona and using reactivity to the 30- and 16-kD bands as the criterion for a positive test.

  19. TSE strain differentiation in mice by immunohistochemical PrP(Sc) profiles and triplex Western blot.

    Science.gov (United States)

    van Keulen, Lucien J M; Langeveld, Jan P M; Dolstra, Corry H; Jacobs, Jorg; Bossers, Alex; van Zijderveld, Fred G

    2015-10-01

    TSE strains are routinely identified by their incubation period and vacuolation profile in the brain after intracerebral inoculation and serial passaging in inbred mouse lines. There are some major drawbacks to this method that are related to the variation in vacuolation that exists in the brains of mice infected with the same TSE strain and to variation between observers and laboratories in scoring vacuolation and determining the final incubation period. We investigated the potential of PrP(Sc) immunohistochemistry and triplex Western blotting as possible alternative methods to differentiate between TSE strains. TSE reference strains ME7, 87A/87V, 22A/22C, 79A/79V and 301C/301V were intracerebrally inoculated in RIII or VM inbred mice that differ in their PrP genotype. Immunohistochemical PrP(Sc) profiles were drawn up by scanning light microscopy both on coronal and sagittal sections. On the basis of the localization of PrP(Sc) in the cerebral cortex, hippocampus, and cerebellar cortex and the overall type of PrP(Sc) staining, all TSE strains could be well differentiated from each other through their typical strain dependent characteristics. In addition, Western blot showed that the combination of glycosylation profile and 12B2 epitope content of PrP(Sc) allowed to distinguish between all reference strains except for ME7 and 22A in VM mice. TSE strains in mice can be identified on the basis of their PrP(Sc) profile alone. The potential to identify TSE strains in ruminants with these PrP(Sc) profiles after a single primary passage in mice will be the topic of future studies. © 2014 British Neuropathological Society.

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

    Science.gov (United States)

    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...

  1. [Clinical manifestation of Lyme borreliosis in children with positive and negatiwe western blot results].

    Science.gov (United States)

    Ołdak, Elzbieta; Rozkiewicz, Doroto; Sulik, Artur

    2008-01-01

    In the afforested area of North-Eastern Poland the risk of Borrelia burgdorferi infection seems to be higher compared to the other regions. Because of unspecific clinical manifestation of Lyme borreliosis in children the positive ELISA IgM results should be confirmed with Western blot IgM tests. Retrospective analysis of clinical signs and symptoms of Lyme borreliosis in children with positive ELISA IgM and positive Western blot IgM results and in children with positive ELISA IgM and negative Western blot IgM results. The study included 20 children reactive with ELISA IgM (Bellco Biomedica, Austria), hospitalized in Pediatric Infectious Diseases Clinic in 2007 due to probable diagnosis of Lyme disease. All children were tested with B. burgdorferi Western blot IgM and/or IgG assay (DRG, Diagnostics, Germany) as a second-step diagnosis. In 10 (50% females, 50% males) out of 20 children the results were positive (borreliosis) and in other 10 (80% females, 20% males) the results were negative (controls). In both groups of patients the retrospective analysis of signs and symptoms was done. The most often clinical manifestation of Lyme borreliosis in children was neuroborreliosis. Children presented Lyme meningitis (30%), facial nerve palsy (10%) and chronic or recurrent headaches (40%), associated with vertigo (20%), weakness (30%), fever (40%), and fatigue syndrome (30%). One patient presented Lyme arthritis. Children of control group presented with unspecific symptoms like isolated headaches (40%), arthralgias (70%), myalgias (10%) and abdomen pain (20%) (1) The most frequent clinical presentation of Lyme borreliosis in analyzed children was neuroborreliosis; (2) Isolated arthralgias in children reactive with B. burgdorferi ELISA IgM need to be confirmed with Western blot assay before implementing the antibiotic therapy.

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

    Science.gov (United States)

    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. © 2015 The International Union of Biochemistry and Molecular Biology.

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

    OpenAIRE

    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, geographi...

  4. Positive IgG Western Blot for Borrelia burgdorferi in Colombia

    Directory of Open Access Journals (Sweden)

    Palacios Ricardo

    1999-01-01

    Full Text Available In order to evaluate the presence of specific IgG antibodies to Borrelia burgdorferi in patients with clinical manifestations associated with Lyme borreliosis in Cali, Colombia, 20 serum samples from patients with dermatologic signs, one cerebrospinal fluid (CSF sample from a patient with chronic neurologic and arthritic manifestations, and twelve serum samples from individuals without clinical signs associated with Lyme borreliosis were analyzed by IgG Western blot. The results were interpreted following the recommendations of the Centers for Diseases Control and Prevention (CDC for IgG Western blots. Four samples fulfilled the CDC criteria: two serum specimens from patients with morphea (localized scleroderma, the CSF from the patient with neurologic and arthritic manifestations, and one of the controls. Interpretation of positive serology for Lyme disease in non-endemic countries must be cautious. However these results suggest that the putative "Lyme-like" disease may correlate with positivity on Western blots, thus raising the possibility that a spirochete genospecies distinct from B. burgdorferi sensu stricto, or a Borrelia species other than B. burgdorferi sensu lato is the causative agent. Future work will focus on a survey of the local tick and rodent population for evidence of spirochete species that could be incriminated as the etiologic agent.

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

    Directory of Open Access Journals (Sweden)

    de Souza Marcilia MA

    2011-01-01

    Full Text Available Abstract 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.

  6. Investigation of the effects of experimental autolysis on the detection of abnormal prion protein in lymphoid and central nervous system tissues from elk and sheep using the Western blotting method.

    Science.gov (United States)

    Huang, Hongsheng; Soutyrine, Andrei; Rendulich, Jasmine; O'Rourke, Katherine; Balachandran, Aru

    2011-01-01

    Tissues unsuitable for standard immunohistochemical and histopathological examinations for chronic wasting disease (CWD) in cervids and for scrapie in sheep are frequently submitted for testing. This study investigated the effects of experimental autolysis on the detection of abnormal prion protein (PrPsc) in lymphoid and central nervous system (CNS) tissues from elk and sheep. The PrPsc was detected using a Western blotting (WB) test following PrPsc enrichment using sodium phosphotungstic acid (PTA) precipitation (PTA-WB). A commercial enzyme-linked immunosorbent assay (ELISA) was used as a reference test for quantitative measurement. This study showed that the amount of PrPsc in lymphoid and CNS tssues from elk and sheep decreased gradually as a result of autolysis, but PrPsc was still detectable after 5 and 15 d incubation at 37°C by PTA-WB for all lymphoid and CNS samples. The results of the ELISA supported those of PTA-WB, particularly for CNS tissues. In conclusion, autolysis at 37°C for 15 d would not significantly affect the detection of PrPsc in lymphoid and CNS tissues by WB and ELISA and, particularly, PTA-WB is a valuable and alternative confirmatory test to detect PrPsc in autolyzed lymphoid and CNS samples.

  7. A single-step simultaneous protein staining procedure for polyacrylamide gels and nitrocellulose membranes by Alta during western blot analysis.

    Science.gov (United States)

    Pal, Jayanta K; Berwal, Sunil K; Soni, Rupali N

    2012-01-01

    A simple method for staining of proteins simultaneously on sodium dodecyl sulfate (SDS) polyacrylamide gels and nitrocellulose membranes by Alta during western blot analysis is described. A 5% solution of Alta, a commercially available cosmetic preparation, is added in the upper tank buffer during electrophoresis. On completion of electrophoresis, the gel is washed in distilled water and viewed on a white light plate and a transilluminator to photograph the protein profiles. The gel is processed for western blot transfer of proteins onto a nitrocellulose membrane, and upon completion, the protein profiles on the membrane are viewed and photographed as stated above. The membrane can then be processed for immunostaining as per the standard procedure. Thus, the staining procedure using Alta is simple, rapid (without any need of destaining), and cost-effective.

  8. Evaluation of two commercial systems for automated processing, reading, and interpretation of Lyme borreliosis Western blots.

    Science.gov (United States)

    Binnicker, M J; Jespersen, D J; Harring, J A; Rollins, L O; Bryant, S C; Beito, E M

    2008-07-01

    The diagnosis of Lyme borreliosis (LB) is commonly made by serologic testing with Western blot (WB) analysis serving as an important supplemental assay. Although specific, the interpretation of WBs for diagnosis of LB (i.e., Lyme WBs) is subjective, with considerable variability in results. In addition, the processing, reading, and interpretation of Lyme WBs are laborious and time-consuming procedures. With the need for rapid processing and more objective interpretation of Lyme WBs, we evaluated the performances of two automated interpretive systems, TrinBlot/BLOTrix (Trinity Biotech, Carlsbad, CA) and BeeBlot/ViraScan (Viramed Biotech AG, Munich, Germany), using 518 serum specimens submitted to our laboratory for Lyme WB analysis. The results of routine testing with visual interpretation were compared to those obtained by BLOTrix analysis of MarBlot immunoglobulin M (IgM) and IgG and by ViraScan analysis of ViraBlot and ViraStripe IgM and IgG assays. BLOTrix analysis demonstrated an agreement of 84.7% for IgM and 87.3% for IgG compared to visual reading and interpretation. ViraScan analysis of the ViraBlot assays demonstrated agreements of 85.7% for IgM and 94.2% for IgG, while ViraScan analysis of the ViraStripe IgM and IgG assays showed agreements of 87.1 and 93.1%, respectively. Testing by the automated systems yielded an average time savings of 64 min/run compared to processing, reading, and interpretation by our current procedure. Our findings demonstrated that automated processing and interpretive systems yield results comparable to those of visual interpretation, while reducing the subjectivity and time required for Lyme WB analysis.

  9. Proteínas inmunodominantes de Brucella Melitensis evaluadas por Western Blot

    Directory of Open Access Journals (Sweden)

    Elizabeth Anaya

    1997-01-01

    Full Text Available Se separaron extractos de proteínas totales de Brucella melitensis en gel 15% SDS-PAGE. Su seroreactividad fue analizada por Western Blot con resultados satisfactorios. Para éste propósito sueros controles negativos (n=03, sueros de pacientes con brucelosis (n=34, cólera (n=12, tifoidea (n=02 y tuberculosis (n=02 fueron usados. Esta prueba inmunodiagnóstica detectó bandas seroreactivas altamente específicas (100% correspondientes a 8,14,18, un complejo de 25-48 y 58kDa. La sensibilidad del test fue del 90% usando los sueros antes mencionados.

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

    Science.gov (United States)

    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.

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

    Directory of Open Access Journals (Sweden)

    Nicholas H Ogden

    Full Text Available 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.

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

    Science.gov (United States)

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

    2000-01-01

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

  13. Total protein analysis as a reliable loading control for quantitative fluorescent Western blotting.

    Directory of Open Access Journals (Sweden)

    Samantha L Eaton

    Full Text Available Western blotting has been a key technique for determining the relative expression of proteins within complex biological samples since the first publications in 1979. Recent developments in sensitive fluorescent labels, with truly quantifiable linear ranges and greater limits of detection, have allowed biologists to probe tissue specific pathways and processes with higher resolution than ever before. However, the application of quantitative Western blotting (QWB to a range of healthy tissues and those from degenerative models has highlighted a problem with significant consequences for quantitative protein analysis: how can researchers conduct comparative expression analyses when many of the commonly used reference proteins (e.g. loading controls are differentially expressed? Here we demonstrate that common controls, including actin and tubulin, are differentially expressed in tissues from a wide range of animal models of neurodegeneration. We highlight the prevalence of such alterations through examination of published "-omics" data, and demonstrate similar responses in sensitive QWB experiments. For example, QWB analysis of spinal cord from a murine model of Spinal Muscular Atrophy using an Odyssey scanner revealed that beta-actin expression was decreased by 19.3±2% compared to healthy littermate controls. Thus, normalising QWB data to β-actin in these circumstances could result in 'skewing' of all data by ∼20%. We further demonstrate that differential expression of commonly used loading controls was not restricted to the nervous system, but was also detectable across multiple tissues, including bone, fat and internal organs. Moreover, expression of these "control" proteins was not consistent between different portions of the same tissue, highlighting the importance of careful and consistent tissue sampling for QWB experiments. Finally, having illustrated the problem of selecting appropriate single protein loading controls, we demonstrate

  14. La técnica de Western Blot como criterio de identidad para la vacuna antimeningocócica Men B Western Blot technique as an identity criterion for Men B antimeningococcal vaccine

    Directory of Open Access Journals (Sweden)

    R. Rosario Diéguez Castro

    2009-12-01

    Full Text Available Se desarrolló y validó la técnica de Western Blot aplicada a la vacuna antimeningocócica Men B producida en el Instituto Finlay con el objetivo de demostrar un criterio de identidad. En el estudio de las proteínas antigénicas de la vacuna, P1.15 y P1.4 en vesícula de membrana externa,monograneles y producto final se emplearon en la identificación anticuerpos monoclonales específicos para estas proteínas. Los parámetros desarrollados en la validación de la técnica fueron: especificidad, límite de detección, repetibilidad, precisión intermedia, reproducibilidad y robustez. El método cumplió con los parámetros señalados, por lo que se consideró validado.Western Blot technique was developed and validated, applied to Men B meningococcal vaccine produced in "Carlos J, Finlay" Institute to demonstrate an identity criterion. In study of antigenic proteins of the vaccine, we used P1.15 y P1.4 in vesicle of external membrane, monogranels, and end product to identify the monoclonal antibodies specific of these proteins. Parameters developed in technique validation included: specificity, detection limit, repetition, average accuracy, reproduction, and strength. Method fulfilled with specified parameters, thus considering its validation.

  15. Profiling EGFR activity in head and neck squamous cell carcinoma by using a novel layered membrane Western blot technology.

    Science.gov (United States)

    Patel, Vyomesh; Ramesh, Arun; Traicoff, June L; Baibakov, Galina; Emmert-Buck, Michael R; Gutkind, J Silvio; Knezevic, Vladimir

    2005-05-01

    Given the role of epidermal growth factor receptor (EGFR) in head and neck squamous cell carcinomas (HNSCC), several rational approaches have now been utilized to abrogate tyrosine kinase activity and its disengagement from downstream signal transducers. Monitoring the activity of these molecules could potentially be useful to determine not only drug efficacy but also to identify HNSCC patients most likely to benefit from this type of therapy. In this study we have used a novel high throughput multi-layered Western blotting (MLWestern) method that allows the detection of multiple proteins from a single experiment in order to characterize key components in the EGFR signaling pathway in HNSCC cells. Total and activated forms of EGFR and the downstream effectors, Erk and Akt were readily detected in HNSCC cells, where in the control cells (HaCaT) these proteins could only be detected in EGF stimulated cells. Results from conventional Western blot and MLWestern were comparable. Clustering analysis of protein expression revealed similarities in cellular response between some of the cell lines indicative of similarities in their biological response. The data indicate that MLWestern can be potentially applied to identify molecular targets that could be used for rational therapeutic intervention strategies.

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

    Energy Technology Data Exchange (ETDEWEB)

    Kale, Sonia [Agharkar Research Institute (India); Kale, Anup [University of Alabama, Center for Materials for Information Technology (United States); Gholap, Haribhau; Rana, Abhimanyu [National Chemical Laboratory, Physical and Materials Chemistry Division (India); Desai, Rama [National Centre for Cell Science (India); Banpurkar, Arun [University of Pune, Department of Physics (India); Ogale, Satishchandra, E-mail: sb.ogale@ncl.res.in [National Chemical Laboratory, Physical and Materials Chemistry Division (India); Shastry, Padma, E-mail: padma@nccs.res.in [National Centre for Cell Science (India)

    2012-03-15

    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 {beta} 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. Comparison of Multispot EIA with Western blot for confirmatory serodiagnosis of HIV.

    Science.gov (United States)

    Torian, Lucia V; Forgione, Lisa A; Punsalang, Amado E; Pirillo, Robert E; Oleszko, William R

    2011-12-01

    Recent improvements in the sensitivity of immunoassays (IA) used for HIV screening, coupled with increasing recognition of the importance of rapid point-of-care testing, have led to proposals to adjust the algorithm for serodiagnosis of HIV so that screening and confirmation can be performed using a dual or triple IA sequence that does not require Western blotting for confirmation. One IA that has been proposed as a second or confirmatory test is the Bio-Rad Multispot(®) Rapid HIV-1/HIV-2 Test. This test would have the added advantage of differentiating between HIV-1 and HIV-2 antibodies. To compare the sensitivity and type-specificity of an algorithm combining a 3rd generation enzyme immunoassay (EIA) followed by a confirmatory Multispot with the conventional algorithm that combines a 3rd generation EIA (Bio-Rad GS HIV-1/HIV-2 Plus O EIA) followed by confirmatory Western blot (Bio-Rad GS HIV-1 WB). 8760 serum specimens submitted for HIV testing to the New York City Public Health Laboratory between May 22, 2007, and April 30, 2010, tested repeatedly positive on 3rd generation HIV-1-2+O EIA screening and received parallel confirmatory testing by WB and Multispot (MS). 8678/8760 (99.1%) specimens tested WB-positive; 82 (0.9%) tested WB-negative or indeterminate (IND). 8690/8760 specimens (99.2%) tested MS-positive, of which 14 (17.1%) had been classified as negative or IND by WB. Among the HIV-1 WB-positive specimens, MS classified 26 (0.29%) as HIV-2. Among the HIV-1 WB negative and IND, MS detected 12 HIV-2. MS detected an additional 14 HIV-1 infections among WB negative or IND specimens, differentiated 26 HIV-1 WB positives as HIV-2, and detected 12 additional HIV-2 infections among WB negative/IND. A dual 3rd generation EIA algorithm incorporating MS had equivalent HIV-1 sensitivity to the 3rd generation EIA-WB algorithm and had the added advantage of detecting 12 HIV-2 specimens that were not HIV-1 WB cross-reactors. In this series an algorithm using EIA

  18. Identification of toxocara canis antigens by Western blot in experimentally infected rabbits

    Directory of Open Access Journals (Sweden)

    MORALES Olga Lucía

    2002-01-01

    Full Text Available Toxocariasis is a frequent helminthiasis that can cause visceral and ocular damage in humans specially in children. The identification of specific antigens of Toxocara canis is important in order to develop better diagnostic techniques. Ten rabbits were infected orally with a dose of 5000 Toxocara canis embryonated eggs. Rabbits were bled periodically and an ELISA assay was performed to determine levels of specific Toxocara IgG antibodies. ELISA detected antibodies at day 15 after infection. Western blot (WB assay was performed using excretory/secretory antigens (E/S of T. canis second stage larvae. Different antigen concentrations were evaluated: 150, 200, 250 and 300 µg/mL. The concentration of 250 µg/mL was retained for analysis. Rabbit sera were diluted 1:100. Secondary antibody was used at a dilution of 1:1000. Results of WB indicated that in the first month after infection specific antibodies against the 200 KDa, 116 KDa, 92 KDa and 35 KDa antigens were detected; antibodies against the 92 KDa, 80 KDa, 66 KDa, 45 KDa, 31 KDa and 28 KDa antigens appeared later. All positive sera in the ELISA test were also positive in WB. Two antigen bands, 92 KDa and 35 KDa, were identified since the beginning and throughout the course of infection. These antigens merit further evaluation as candidates for use in diagnosis.

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

    Science.gov (United States)

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

    1996-05-01

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

  20. Indeterminate human immunodeficiency virus western blot results in Iranian patients with discordant screening assay results

    International Nuclear Information System (INIS)

    Ravanshad, M.; Sabahi, F.; Mahboudi, F.; Sabahi, F.

    2006-01-01

    The Western blot (WB) assay is the most widely accepted confirmatory assay for the detection and confirmation of antibodies to human immunodeficiency virus type 1 (HIV-1) and 2 (HIV-2). However, indeterminate WB reactivity to HIV-1 and HIV-2 proteins may occur in individuals who do not appear to be infected with HIV. In this study, we describe the results of indeterminate WB reactivity in Iranian patients with discordant screening assays. The samples were obtained from Iranian Blood Transfusion Center, Tehran, Iran and evaluated in the Biotechnology Process Development Center, Pasteur Institute of Iran, Tehran, Iran between 2003 and 2004. A total of 4707 were tested for the presence of HIV-1 antibodies. Six hundred and four (12.8%) patients tested for HIV were positive for HIV-1 antibody. Nine (1.49%) have discordant results among screening assays and indeterminate WB results as interpreted by Centers for Disease Control and Prevention (CDC) criteria. Most (66.7%) of these indeterminate WB results were due to p24 reactivity. However, 2(22.2%) display reactivity to both gp41 and gp120 proteins [Positive by World Health Organization (WHO) criteria]. Of 9 WB assays initially indeterminate by the CDC criteria and with follow-up samples 8(88.8%) became negative when retested subsequently while one (11.1%) remained indeterminate for more than a year and were thus considered negative. In addition all the indeterminate samples were negative when assessed by polymerase chain reaction assay. In general, there were was an 88.8% concordance between the CDC and WHO criteria for an indeterminate WB result. The CDC II criteria for an indeterminate WB result. The CDC II criteria best met the specified objectives for diagnosis in our setting. (author)

  1. [Western Blot diagnostic yield for simultaneous antibody-detection in patients with human cysticercosis, hydatidosis, and human fascioliasis].

    Science.gov (United States)

    Davelois, Kelly; Escalante, Hermes; Jara, César

    2016-01-01

    . To determine the diagnostic yield using western blotting to simultaneously detect antibodies in patients with human cysticercosis, hydatidosis, and human fascioliasis. Materials and methods . Cross-sectional study of diagnostic yield assessment. Excretory/secretory antigens were obtained from Taenia solium larvae, Echinococcus granulosus cysts, and the adult flukes of Fasciola hepática, which were then separated using the polyacrylamide gel electrophoresis technique, transferred, and attached to a nitrocellulose membrane to be probed with sera from the patient infected with the three parasites. The sensitivity of the technique was assessed using 300 individual serum samples, 60 pools of two parasites, and 20 pools of three parasites with 75 sera from patients with other parasites, 10 from patients with other diseases, and 15 from patients without parasites. Results . The technique revealed 13 glycoproteins (GP): GP 35, 31, 24, 23, 18, 17, 14, and 13 kDa for cysticercosis; GP 8, 16, and 21 kDa for hydatidosis; and GP 17 and 23 kDa for fascioliasis. The test detected the presence of antibodies with a sensitivity of 96% (95% confidence interval [CI] = 94.62-98.54%) in the detection of one or the thirteen bands, a specificity of 100% (95% CI = 99.50-100.00%); individually, there was a sensitivity for cysticercosis of 97% (95% CI = 93.16-100.00%), for hydatidosis of 94% (95% CI = 88.85-99.15%) and for fascioliasis of 96% (95% CI = 91.66-100.00%). Conclusions . Western blotting is effective in the simultaneous detection of antibodies in patients with human cysticercosis, hydatidosis, and fascioliasis, and it can be used as a diagnostic test to either rule out or confirm the presence of antibodies in endemic areas.

  2. In Situ Blotting : A Novel Method for Direct Transfer of Native Proteins from Sectioned Tissue to Blotting Membrane

    NARCIS (Netherlands)

    Okabe, Masashi; Nyakas, Csaba; Buwalda, Bauke; Luiten, Paul G.M.

    1993-01-01

    We describe a novel technique for direct transfer of native proteins from unfixed frozen tissue sections to an immobilizing matrix, e.g., nitrocellulose, polyvinyliden difluoride, or positively charged nylon membranes. Proteins are directly blotted onto the membrane, providing optimal accessibility

  3. The combination of quantitative PCR and western blot detecting CP4-EPSPS component in Roundup Ready soy plant tissues and commercial soy-related foodstuffs.

    Science.gov (United States)

    Xiao, Xiao; Wu, Honghong; Zhou, Xinghu; Xu, Sheng; He, Jian; Shen, Wenbiao; Zhou, Guanghong; Huang, Ming

    2012-06-01

    With the widespread use of Roundup Ready soy (event 40-3-2) (RRS), the comprehensive detection of genetically modified component in foodstuffs is of significant interest, but few protein-based approaches have been found useful in processed foods. In this report, the combination of quantitative PCR (qPCR) and western blot was used to detect cp4-epsps gene and its protein product in different RRS plant tissues and commercial soy-containing foodstuffs. The foods included those of plant origin produced by different processing procedures and also some products containing both meat and plant protein concentrates. The validity of the 2 methods was confirmed first. We also showed that the CP4-EPSPS protein existed in different RRS plant tissues. In certain cases, the results from the western blot and the qPCR were not consistent. To be specific, at least 2 degraded fragments of CP4-EPSPS protein (35.5 and 24.6 kDa) were observed. For dried bean curd crust and deep-fried bean curd, a degraded protein fragment with the size of 24.6 kDa appeared, while cp4-epsps gene could not be traced by qPCR. In contrast, we found a signal of cp4-epsps DNA in 3 foodstuffs, including soy-containing ham cutlet product, meat ball, and sausage by qPCR, while CP4-EPSPS protein could not be detected by western blot in such samples. Our study therefore concluded that the combination of DNA- and protein-based methods would compensate each other, thus resulting in a more comprehensive detection from nucleic acid and protein levels. The combination of quantitative PCR (qPCR) and western blot was used to detect cp4-epsps gene and its protein product in different Roundup Ready soy (event 40-3-2) plant tissues and commercial soy-containing foodstuffs. The foods included those of plant origin produced by different processing procedures and also some products containing a combination of both meat and plant protein concentrates. This study indicated that the combination of DNA- and protein-based methods

  4. Western blot banding pattern in early Lyme borreliosis among patients from an endemic region of north-eastern Poland.

    Science.gov (United States)

    Flisiak, R; Wierzbicka, I; Prokopowicz, D

    1998-01-01

    Aim of this study was evaluation of Western blot banding patterns in different clinical forms of early Lyme borreliosis diagnosed in patients from north-eastern Poland, recognized as endemic for tick-borne diseases. Study was performed on serum samples of 48 patients with Lyme borreliosis and 26 healthy volunteers, as controls. Samples tested routinely for total antibody with enzyme immunoassay were subsequently analysed for specific antibodies with Western blot based on antigen extract of European strain of Borrelia burgdorferi. In patients, IgM antibodies were the most frequently directed against 41 kDa and 58 kDa antigens, whereas in control group only antibodies against 45 kDa and 58 kDa were present. Similar response was observed in respect to IgG antibodies. Evaluation of banding pattern in respect to clinical form of the disease revealed the highest prevalence of IgM and IgG anti-41 kDa antibodies in patients with erythema migrans and Lyme arthritis, and anti-58 kDa in neuroborreliosis patients, who had no anti-21 kDa antibodies. Relatively high frequency of IgG antibodies against 21, 30 and 93 kDa antigens was typical for neuroborreliosis. Bands count was significantly higher in different clinical forms of the disease than in controls, and it was the highest in neuroborreliosis. Combined analysis of Western blot results (IgM/IgG) enabled to achieve higher sensitivity (84%) and specificity (100%) than available with the most recommended EIA kits.

  5. Detection of Potentially Diagnostic Leishmania Antigens with Western Blot Analysis of Sera from Patients with Cutaneous and Visceral Leishmaniases

    Directory of Open Access Journals (Sweden)

    Seyyed Javad SEYYEDTABAEI

    2017-06-01

    Full Text Available Background: Visceral leishmaniasis (VL and cutaneous leishmaniasis (CL are important public health problems in Iran. We aimed to evaluate the diagnostic potential of Western blot (WB compared with indirect immunofluorescence test (IFAT to serodiagnosis of leishmaniasis.Methods: This study was performed from 2010-2014 and participants were different parts of Iran. Serum samples were obtained from 43 patients with proven CL, 33 patients with proven VL, 39 patients with other parasitic diseases and 23 healthy individuals. Results: WB sensitivity for CL and VL was 100% and 91%, compared to IFA 4.6% and 87.8%, respectively. Sera from patients with CL and VL recognized numerous antigens with molecular weights ranging from 14 to 68 kDa and 12 to 94 kDa, respectively. The most sensitive antigens were 14 and 16 kDa for CL recognized by 100% of the sera from patients with proven CL and 12, 14 and 16 kDa for VL, recognized by 63.6%, 100% and 63.6% of the sera from patients with proven VL respectively. WB analysis is more sensitive than IFAT for the diagnosis of leishmaniasis particularly in cases of cutaneous leishmaniasis. The 12, 14 and 16 kDa can be valuable diagnostic molecules for serodiagnosis of leishmaniasis because at least two immunogenic molecules were simultaneously detected by all patient sera, as well as produced antibodies against these antigens have no cross-reactivity with other control groups.Conclusion: WB could be useful for screening and serodiagnosis of CL and VL in epidemiologic studies in endemic areas.

  6. Southern blotting.

    Science.gov (United States)

    Brown, T

    2001-05-01

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

  7. Glycophospholipid Formulation with NADH and CoQ10 Significantly Reduces Intractable Fatigue in Western Blot-Positive ‘Chronic Lyme Disease’ Patients: Preliminary Report

    Directory of Open Access Journals (Sweden)

    Garth L. Nicolson

    2012-03-01

    Full Text Available Background: An open label 8-week preliminary study was conducted in a small number of patients to determine if a combination oral supplement containing a mixture of phosphoglycolipids, coenzyme Q10 and microencapsulated NADH and other nutrients could affect fatigue levels in long-term, Western blot-positive, multi-symptom ‘chronic Lyme disease’ patients (also called ‘post-treatment Lyme disease’ or ‘post Lyme syndrome’ with intractable fatigue. Methods: The subjects in this study were 6 males (mean age = 45.1 ± 12.4 years and 10 females (mean age = 54.6 ± 7.4 years with ‘chronic Lyme disease’ (determined by multiple symptoms and positive Western blot analysis that had been symptomatic with chronic fatigue for an average of 12.7 ± 6.6 years. They had been seen by multiple physicians (13.3 ± 7.6 and had used many other remedies, supplements and drugs (14.4 ± 7.4 without fatigue relief. Fatigue was monitored at 0, 7, 30 and 60 days using a validated instrument, the Piper Fatigue Scale.Results: Patients in this preliminary study responded to the combination test supplement, showing a 26% reduction in overall fatigue by the end of the 8-week trial (p< 0.0003. Analysis of subcategories of fatigue indicated that there were significant improvements in the ability to complete tasks and activities as well as significant improvements in mood and cognitive abilities. Regression analysis of the data indicated that reductions in fatigue were consistent and occurred with a high degree of confidence (R2= 0.998. Functional Foods in Health and Disease 2012, 2(3:35-47 Conclusions: The combination supplement was a safe and effective method to significantly reduce intractable fatigue in long-term patients with Western blot-positive ‘chronic Lyme disease.’

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

    Science.gov (United States)

    Kurien, Biji T; Scofield, R Hal

    2015-01-01

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

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

    Science.gov (United States)

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

    2016-06-14

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

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

    Directory of Open Access Journals (Sweden)

    Luiz Claudio Pereira Ribeiro

    2013-09-01

    Full Text Available 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.

  11. Diagnostic potential of Western blot analysis of sera from dogs with leishmaniasis in endemic areas and significance of the pattern.

    Science.gov (United States)

    Aisa, M J; Castillejo, S; Gallego, M; Fisa, R; Riera, M C; de Colmenares, M; Torras, S; Roura, X; Sentis, J; Portus, M

    1998-02-01

    Serum samples collected from 237 dogs in Catalonia (northeastern Spain) were screened by Western blot analysis to detect the presence of antibodies specific to different Leishmania infantum polypeptide fractions. Leishmaniasis was confirmed in 72 of these dogs by direct examination and/or culture. Another 165 animals from the Priorat region were studied periodically for 2-8 years between 1987 and 1995, giving a total of 565 determinations. A control group of 93 dogs from nonendemic areas was also studied. Sera from dogs with leishmaniasis recognized antigens with molecular weights ranging from 12 to 85 kD. The most sensitive antigens were those of 70, 65, 46, 30, 28, 14, and 12 kD, which were recognized by 75%, 75%, 78%, 75%, 81%, 79%, and 75%, respectively, of the sera from dogs with positive parasitologic examination results. Antigens of 70 and 65 kD were also recognized by two dogs from nonendemic areas. Antigens of 14 and 12 kD were the first to be recognized by sera of asymptomatic dogs with titers less than the cut-off value of the dot-ELISA that increased during the longitudinal study, and the presence of antibodies specific for these fractions was observed for up to six years before seroconversion observed by dot-ELISA. These antibodies were also the first to disappear in dogs in which the disease was self-limited. The study corroborates the high sensitivity and specificity of Western blots in the diagnosis of canine leishmaniasis when the bands of low molecular weight (less than 46 kD) are considered, and indicates that fractions of 14 and 12 kD are useful in detecting early forms of the disease.

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

    Directory of Open Access Journals (Sweden)

    Marzieh ASHRAFMANSOURI

    2015-12-01

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

  13. Standardization of micro-enzyme-linked immunosorbent assay (ELISA) and Western blot for detection of Trypanosoma cruzi antibodies using extracts from Mexican strains as antigens.

    Science.gov (United States)

    Sánchez, B; Monteón, V; Reyes, P A; Espinoza, B

    2001-01-01

    This report describes two assays for the detection of anti-Trypanosoma cruzi antibodies using Mexican strains of the parasite and the concordance with two assays previously evaluated at the Instituto Nacional de Cardiología Ignacio Chávez in Mexico City. Micro-enzyme-linked immunosorbent assay (ELISA) and Western blot were used for the detection of T. cruzi antibodies with a total extract of epimastigote from Ninoa and Queretaro, which are Mexican strains of T. cruzi. To standardize these methods, a total of 246 serum samples was used. In addition, sera from six confirmed Mexican chronic individuals in the asymptomatic phase were also used for comparison with the Argentinean antigen. ELISA was 100% specific in that no false positive results were found with sera of both healthy individuals and non-Chagasic cardiopaths. Sera from individuals infected with Leishmania sp. showed approximately 16% of cross-reaction with ELISA. The test showed a positive predictive value of 90% and a negative predictive value of 100%. Western blot was also a highly sensitive test for detecting chronic Chagasic symptomatic patients from Mexico because no false negative results were obtained. Furthermore, it was possible to use Western blot to detect seven immunodominant antigens of approximately 30, 32, 40, 42, 65, 70, and 83 kDa. Concordance with two previous standardized tests at the Instituto Nacional de Cardiología showed a Kappa index of 0.96, indicating high concordance between the results obtained at these two laboratories. Finally, ELISA using Ninoa antigen extract was more sensitive than ELISA with an Argentinean extract, which failed to detect individuals in the chronic asymptomatic phase (undetermined phase) of infection. This study indicates that ELISA and Western blot using Ninoa and/or Queretaro extracts of T. cruzi as antigens are useful tools in the detection of individuals who have been exposed to T. cruzi both in the undetermined/asymptomatic and symptomatic phases

  14. The early days of blotting.

    Science.gov (United States)

    Southern, Edwin

    2015-01-01

    The history of the development of DNA blotting is described in this chapter. DNA blotting, involving the transfer of electrophoretically separated DNA fragments to a membrane support through capillary action, is also known as Southern blotting. This procedure enables the detection of a specific DNA sequence by hybridization with probes. The term Southern blotting led to a "geographic" naming tradition, with RNA blotting bearing the name Northern blotting and protein transfer to membranes becoming known as Western blotting.

  15. Demonstration of monoclonal anti-carcinoembryonic antigen (CEA) antibody internalization by electron microscopy, western blotting and radioimmunoassay.

    Science.gov (United States)

    Tsaltas, G; Ford, C H; Gallant, M

    1992-01-01

    One of the important factors affecting the action of monoclonal antibodies (Mabs) or immunoconjugates on tumour sites depends on whether the Mab is internalized by the cancer cells in question. The underexplored subject of internalization is discussed in this paper, and a number of in vitro techniques for investigating internalization are evaluated, using a model which consists of a well characterized anti-carcinoembryonic antigen (anti-CEA) Mab and a number of CEA expressing human cancer cell lines. Employing two alternative radiolabeling assays, evidence for internalization of the anti-CEA Mab by a CEA-positive colorectal cancer cell line (LS174T) was obtained throughout the time intervals examined (5 min to 150 min). Electronmicroscopy employing horseradish-peroxidase labeled anti-CEA Mab and control antibody permitted direct visualization of anti-CEA Mab-related staining in intracellular compartments of a high CEA-expressor human colorectal cell line (SKCO1). Finally Western blots of samples derived from cytosolic and membrane components of solubilized cells from lung and colonic cancer cell lines provided evidence for internalized anti-CEA Mab throughout seven half hour intervals, starting at 5 minutes. Internalized anti-CEA was detected in all CEA expressing cell lines (LS174T, SKCO1, BENN) but not in the case of a very low CEA expressor line (COLO 320).

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

    Science.gov (United States)

    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. Copyright © 2011 Elsevier B.V. All rights reserved.

  17. Cross-Reactions between Toxocara canis and Ascaris suum in the diagnosis of visceral larva migrans by western blotting technique

    Directory of Open Access Journals (Sweden)

    NUNES Cáris Maroni

    1997-01-01

    Full Text Available Visceral larva migrans (VLM is a clinical syndrome caused by infection of man by Toxocara spp, the common roundworm of dogs and cats. Tissue migration of larval stages causes illness specially in children. Because larvae are difficult to detect in tissues, diagnosis is mostly based on serology. After the introduction of the enzyme-linked immunosorbent assay (ELISA using the larval excretory-secretory antigen of T. canis (TES, the diagnosis specificity was greatly improved although cross-reactivity with other helminths are still being reported. In Brazil, diagnosis is routinely made after absorption of serum samples with Ascaris suum antigens, a nematode antigenicaly related with Ascaris lumbricoides which is a common intestinal nematode of children. In order to identify T. canis antigens that cross react to A. suum antigens we analyzed TES antigen by SDS-PAGE and Western blotting techniques. When we used serum samples from patients suspected of VLM and positive result by ELISA as well as a reference serum sample numerous bands were seen (molecular weight of 210-200 kDa, 116-97 kDa, 55-50 kDa and 35-29 kDa. Among these there is at least one band with molecular weight around 55-66 kDa that seem to be responsible for the cross-reactivity between T. canis e A. suum once it disappears when previous absorption of serum samples with A. suum antigens is performed

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

    International Nuclear Information System (INIS)

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

    1982-01-01

    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

  19. Detection of proteins on blot transfer membranes.

    Science.gov (United States)

    Sasse, Joachim; Gallagher, Sean R

    2003-11-01

    In the basic and alternate protocols of this unit, proteins are stained after electroblotting from polyacrylamide gels to blot transfer membranes. If the samples of interest are electrophoresed in duplicate and transferred to a blot transfer membrane, half of the membrane can be stained to determine the efficiency of transfer to the membrane and the other half can be used for immunoblotting (i.e., western blotting). Detection limits of each staining method are given along with a list of compatible blot transfer membranes and gels. A support protocol describes a method for alkali treatment that enhances subsequent staining of bound proteins.

  20. Characterization of Sm14 related components in different helminths by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting analysis

    Directory of Open Access Journals (Sweden)

    Nilton Thaumaturgo

    2002-10-01

    Full Text Available Sm14 was the first fatty acid-binding protein homologue identified in helminths. Thereafter, members of the same family were identified in several helminth species, with high aminoacid sequence homology between them. In addition, immune crossprotection was also reported against Fasciola hepatica infection, in animals previously immunized with the Schistosoma mansoni vaccine candidate, r-Sm14. In the present study, data on preliminary sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting analysis of nine different helminth extracts focusing the identification of Sm14 related proteins, is reported. Out of these, three extracts - Ascaris suum (males and females, Echinostoma paraensei, and Taenia saginata - presented components that comigrated with Sm14 in SDS-PAGE, and that were recognized by anti-rSm14 policlonal serum, in Western blotting tests.

  1. Development of EMab-51, a Sensitive and Specific Anti-Epidermal Growth Factor Receptor Monoclonal Antibody in Flow Cytometry, Western Blot, and Immunohistochemistry.

    Science.gov (United States)

    Itai, Shunsuke; Kaneko, Mika K; Fujii, Yuki; Yamada, Shinji; Nakamura, Takuro; Yanaka, Miyuki; Saidoh, Noriko; Handa, Saori; Chang, Yao-Wen; Suzuki, Hiroyoshi; Harada, Hiroyuki; Kato, Yukinari

    2017-10-01

    The epidermal growth factor receptor (EGFR) is a member of the human epidermal growth factor receptor (HER) family of receptor tyrosine kinases and is involved in cell growth and differentiation. EGFR homodimers or heterodimers with other HER members, such as HER2 and HER3, activate downstream signaling cascades in many cancers. In this study, we developed novel anti-EGFR monoclonal antibodies (mAbs) and characterized their efficacy in flow cytometry, Western blot, and immunohistochemical analyses. First, we expressed the full-length or ectodomain of EGFR in LN229 glioblastoma cells and then immunized mice with LN229/EGFR or ectodomain of EGFR, and performed the first screening using enzyme-linked immunosorbent assays. Subsequently, we selected mAbs according to their efficacy in flow cytometry (second screening), Western blot (third screening), and immunohistochemical (fourth screening) analyses. Among 100 mAbs, only one clone EMab-51 (IgG 1 , kappa) reacted with EGFR in Western blot analysis. Finally, immunohistochemical analyses with EMab-51 showed sensitive and specific reactions against oral cancer cells, warranting the use of EMab-51 to detect EGFR in pathological analyses of EGFR-expressing cancers.

  2. Estandarización de la técnica de Western blot para el diagnóstico de la fasciolosis humana utilizando antígenos de excreción-secreción de Fasciola hepática Western blot technique standardization of the diagnosis of human fasciolosis using Fasciola hepatica excreted-secreted antigens

    Directory of Open Access Journals (Sweden)

    Hermes Escalante

    2011-09-01

    Full Text Available Objetivos. Evaluar la eficacia de la técnica de electroinmunotransferencia (EITB o Western blot utilizando antígenos de excreción-secreción de las formas adultas de Fasciola hepatica (Fh E/S Ag para el diagnóstico de la fasciolosis humana. Materiales y métodos. Los antígenos fueron obtenidos a las 18 horas de incubación en medio Minimum Essential Eagle y preparados a la concentración proteica de 0,15 ug/uL; los cuales, al ser enfrentados con un pool de sueros de pacientes con fasciolosis confirmada por el hallazgo de huevos del parásito en las heces, se detectaron los antígenos de 10, 12, 17, 23, 27, 30, 36, 43, 66 y 136 KDa, con los cuales se desarrolló la técnica de Western blot. La sensibilidad se evaluó empleando sueros de 67 pacientes con fasciolosis, y la especificidad con sueros de 57 pacientes con otras parasitosis y diez sueros de personas no parasitadas. Resultados. De los 67 sueros, 64 reaccionaron con la banda de 23 KDa y 61 con la banda de 17KDa. Estas dos bandas no fueron detectadas por ninguno de los sueros de pacientes con otras parasitosis, ni de personas no parasitadas, siendo por ello consideradas como específicas y diagnósticas. Conclusiones. La sensibilidad de la prueba, utilizando las bandas de 17 y 23 KDa, fue de 95,5 % cuando se presenta reacción positiva en una o en las dos bandas, siendo la especificidad para estos dos antígenos de 100 % con un valor predictivo positivo de 100 % y un valor predictivo negativo de 95,71 %.Objectives. To evaluate the performance of the enzyme-linked immunoelectrotransfer blot assay (EITB, Western blot using excretory/secretory antigens from adult forms of Fasciola hepatica (Fh E/S Ag for the diagnosis of human fasciolosis. Materials and methods. Antigens were obtained after 18 hours of incubation in culture medium Minimum Essential Eagle, prepared at a protein concentration of 0.15 ug/uL and run against a pool of sera of patients with proven fasciolosis (confirmed by the

  3. INSITU BLOTTING - A NOVEL METHOD FOR DIRECT TRANSFER OF NATIVE PROTEINS FROM SECTIONED TISSUE TO BLOTTING MEMBRANE - PROCEDURE AND SOME APPLICATIONS

    NARCIS (Netherlands)

    OKABE, M; NYAKAS, C; BUWALDA, B; LUITEN, PGM

    We describe a novel technique for direct transfer of native proteins from unfixed frozen tissue sections to an immobilizing matrix, e.g., nitrocellulose, polyvinyliden difluoride, or positively charged nylon membranes. Proteins are directly blotted onto the membrane, providing optimal accessibility

  4. Protein blotting protocol for beginners.

    Science.gov (United States)

    Petrasovits, Lars A

    2014-01-01

    The transfer and immobilization of biological macromolecules onto solid nitrocellulose or nylon (polyvinylidene difluoride (PVDF)) membranes subsequently followed by specific detection is referred to as blotting. DNA blots are called Southerns after the inventor of the technique, Edwin Southern. By analogy, RNA blots are referred to as northerns and protein blots as westerns (Burnette, Anal Biochem 112:195-203, 1981). With few exceptions, western blotting involves five steps, namely, sample collection, preparation, separation, immobilization, and detection. In this chapter, protocols for the entire process from sample collection to detection are described.

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

    DEFF Research Database (Denmark)

    Albers, Eliene; Sbroggiò, Mauro; Martin Gonzalez, Javier

    2017-01-01

    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...

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

    Science.gov (United States)

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

    2016-08-01

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

  7. Recombinant antigen-based immuno-slot blot method for serodiagnosis of syphilis

    Directory of Open Access Journals (Sweden)

    N.S. Sato

    2004-07-01

    Full Text Available Three recombinant antigens of Treponema pallidum Nichols strain were fused with GST, cloned and expressed in Escherichia coli, resulting in high levels of GST-rTp47 and GST-rTp17 expression, and supplementation with arginine tRNA for the AGR codon was needed to obtain GST-rTp15 overexpression. Purified fusion protein yields were 1.9, 1.7 and 5.3 mg/l of cell culture for GST-rTp47, GST-rTp17 and GST-rTp15, respectively. The identities of the antigens obtained were confirmed by automated DNA sequencing using ABI Prism 310 and peptide mapping by Finningan LC/MS. These recombinant antigens were evaluated by immuno-slot blot techniques applied to 137 serum samples from patients with a clinical and laboratory diagnosis of syphilis (61 samples, from healthy blood donors (50 samples, individuals with sexually transmitted disease other than syphilis (3 samples, and from individuals with other spirochetal diseases such as Lyme disease (20 samples and leptospirosis (3 samples. The assay had sensitivity of 95.1% (95% CI, 86.1 to 98.7% and a specificity of 94.7% (95% CI, 87.0 to 98.7%; a stronger reactivity was observed with fraction rTp17. The immunoreactivity results showed that fusion recombinant antigens based-immuno-slot blot techniques are suitable for use in diagnostic assays for syphilis.

  8. Northern blotting analysis

    DEFF Research Database (Denmark)

    Josefsen, Knud; Nielsen, Henrik

    2011-01-01

    Northern blotting analysis is a classical method for analysis of the size and steady-state level of a specific RNA in a complex sample. In short, the RNA is size-fractionated by gel electrophoresis and transferred by blotting onto a membrane to which the RNA is covalently bound. Then, the membrane...... is analysed by hybridization to one or more specific probes that are labelled for subsequent detection. Northern blotting is relatively simple to perform, inexpensive, and not plagued by artefacts. Recent developments of hybridization membranes and buffers have resulted in increased sensitivity closing...

  9. Testing UK blood donors for exposure to human parvovirus 4 using a time-resolved fluorescence immunoassay to screen sera and Western blot to confirm reactive samples.

    Science.gov (United States)

    Maple, Peter A C; Beard, Stuart; Parry, Ruth P; Brown, Kevin E

    2013-10-01

    Human parvovirus 4 (ParV4), a newly described member of the family Parvoviridae, like B19V, has been found in pooled plasma preparations. The extent, and significance, of ParV4 exposure in UK blood donors remain to be determined and reliable detection of ParV4 immunoglobulin (Ig)G, using validated methods, is needed. With ParV4 virus-like particles a ParV4 IgG time-resolved fluorescence immunoassay (TRFIA) was developed. There is no gold standard or reference assay for measuring ParV4 IgG and the utility of the TRFIA was first examined using a panel of sera from people who inject drugs (PWIDS)--a high-prevalence population for ParV4 infection. Western blotting was used to confirm the specificity of TRFIA-reactive sera. Two cohorts of UK blood donor sera comprising 452 sera collected in 1999 and 156 sera collected in 2009 were tested for ParV4 IgG. Additional testing for B19V IgG, hepatitis C virus antibodies (anti-HCV), and ParV4 DNA was also undertaken. The rate of ParV4 IgG seroprevalence in PWIDS was 20.7% and ParV4 IgG was positively associated with the presence of anti-HCV with 68.4% ParV4 IgG-positive sera testing anti-HCV-positive versus 17.1% ParV4 IgG-negative sera. Overall seropositivity for ParV4 IgG, in 608 UK blood donors was 4.76%. The ParV4 IgG seropositivity for sera collected in 1999 was 5.08%, compared to 3.84% for sera collected in 2009. No ParV4 IgG-positive blood donor sera had detectable ParV4 DNA. ParV4 IgG has been found in UK blood donors and this finding needs further investigation. © 2013 American Association of Blood Banks.

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

    Science.gov (United States)

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

    2015-08-01

    The aim of this study was to investigate the presence of canine immunoglobulins (Ig) G against Demodex proteins in the sera of healthy dogs and of dogs with juvenile generalized demodicosis (CanJGD) with or without secondary pyoderma. Demodex mites were collected from dogs with CanJGD. Protein concentration was measured and a western blot technique was performed. Pooled sera from healthy dogs reacted mainly with antigen bands ranging from 55 to 72 kDa. Pooled sera from dogs with CanJGD without secondary pyoderma reacted either with 10 kDa antigen band or 55 to 72 kDa bands. Pooled sera from dogs with CanJGD with secondary pyoderma reacted only with a 10 kDa antigen band. The results of this study suggest that both healthy dogs and dogs with CanJGD develop a humoral response against different proteins of Demodex canis. Copyright © 2015 Elsevier Ltd. All rights reserved.

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

    Science.gov (United States)

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

    2016-01-01

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

  12. Rendimiento diagnóstico del Western Blot para detectar simultáneamente anticuerpos en pacientes con cisticercosis, hidatidosis y fascioliasis humana

    Directory of Open Access Journals (Sweden)

    Kelly Davelois

    Full Text Available Objetivo. Determinar el rendimiento diagnóstico de la técnica de Western Blot para detectar simultáneamente anticuerpos en pacientes con cisticercosis, hidatidosis y fascioliasis humana. Materiales y métodos. Estudio transversal de evaluación de prueba diagnóstica. Se obtuvieron los antígenos de excreción-secreción de las larvas de Taenia solium, quistes de Echinococcus granulosus; y la forma adulta de Fasciola hepática; que luego fueron separados electroforéticamente en geles de poliacrilamida individuales, transferidos y fijados a una membrana de nitrocelulosa para ser enfrentados con sueros de pacientes con las tres parasitosis. La sensibilidad de la técnica se evaluó empleando 300 sueros individuales, 60 pools de dos parasitosis y 20 pools de tres parasitosis y la especificidad con 75 sueros de pacientes con otras parasitosis, 10 de pacientes con otras enfermedades y 15 sueros de personas no parasitadas. Resultados. La técnica reconoció trece glicoproteínas (GP: GP 35, 31, 24, 23, 18, 17, 14 y 13 kDa para cisticercosis, GP 8,16 y 21 kDa para hidatidosis y GP: 17 y 23 kDa para fascioliasis. La prueba detectó la presencia de anticuerpos alcanzando una sensibilidad de 96% (IC95%: 94,62-98,54% en la detección de una o las trece bandas, una especificidad de 100% (IC95%: 99,50 - 100,00%; individualmente, se tuvo una sensibilidad para cisticercosis de 97% (IC95%: 93,16-100%, para hidatidosis de 94% (IC95%: 88,85-99,15% y para fascioliasis de 96% (IC95%: 91,66-100%. Conclusiones. La prueba de Western blot es eficaz en la detección, simultanea de anticuerpos en pacientes con cisticercosis, hidatidosis y fascioliasis humana, y puede ser utilizada como prueba de descarte o confirmatoria en zonas endémicas.

  13. Western blot seroindeterminate individuals for Human T-lymphotropic Virus 1/2 (HTLV-1/2 in Fortaleza (Brazil: a serological and molecular diagnostic and epidemiological approach

    Directory of Open Access Journals (Sweden)

    Santos Terezinha de Jesus Teixeira

    2003-01-01

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

  14. Western blot seroindeterminate individuals for Human T-lymphotropic Virus 1/2 (HTLV-1/2 in Fortaleza (Brazil: a serological and molecular diagnostic and epidemiological approach

    Directory of Open Access Journals (Sweden)

    Terezinha de Jesus Teixeira Santos

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

  15. Direct visualization of epidermal growth factor receptors (EGFR) in A431 and placental cell membrane by western blot with 125I-EGF

    International Nuclear Information System (INIS)

    Lin, P.H.; Selinfreund, R.; Wharton, W.

    1986-01-01

    Using the western blot technique, they have devised a new procedure that allowed the direct visualization of both the 150KD and the 170KD forms of EGFR by its natural ligand, 125 I-EGF. A431, and placental plasmalemma were purified and solubilized in either SDS-PAGE buffer (without DTT, EDTA) or Triton X-100 (0.5%), resolved on PAGE and electrophoretically transferred onto nitrocellulose (NC) paper. In the absence of boiling, SDS did not denature the EGFR. Although EGER band can be detected after hybridization with 125 I-EGF, the receptor signal was considerably improved with the addition of 0.1% Tween-20. The binding of 125 I-EGF to the both the 150KD and the 170KD bands of the EGFR was specific, reversible and increased with the amount of membrane protein present. The direct visualization of the EGFR using its natural ligand eliminated the necessity for the time consuming antibody preparation. Presently, they are using this technique to identify specific receptors for other ligands

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

    Science.gov (United States)

    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.

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

    Science.gov (United States)

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

    2005-12-01

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

  18. Myostatin inhibitors in sports drug testing: Detection of myostatin-neutralizing antibodies in plasma/serum by affinity purification and Western blotting.

    Science.gov (United States)

    Walpurgis, Katja; Thomas, Andreas; Schänzer, Wilhelm; Thevis, Mario

    2016-02-01

    Myostatin is a key regulator of skeletal muscle growth and inhibition of its signaling pathway results in an increased muscle mass and function. The aim of this study was to develop a qualitative detection assay for myostatin-neutralizing antibodies for doping control purposes by using immunological approaches. To detect different types of myostatin-neutralizing antibodies irrespective of their amino acid sequence, an immunological assay specific for antibodies directed against myostatin and having a human Fc domain was established. Affinity purification and Western blotting strategies were combined to allow extracting and identifying relevant analytes from 200 μL of plasma/serum in a non-targeted approach. The assay was characterized regarding specificity, linearity, precision, robustness, and recovery. The assay was found to be highly specific, robust, and linear from 0.1 to 1 μg/mL. The precision was successfully specified at three different concentrations and the recovery of the affinity purification was 58%. Within this study, an immunological detection assay for myostatin-neutralizing antibodies present in plasma/serum specimens was developed and successfully characterized. The presented approach can easily be modified to include other therapeutic antibodies and serves as proof-of-concept for the detection of antibody-based myostatin inhibitors in doping control samples. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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

    Science.gov (United States)

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

    2013-08-01

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

  20. Evaluación de la técnica Western blot para la detección de antígenos de Hymenolepis nana

    Directory of Open Access Journals (Sweden)

    Flora Chávez-Salas

    2013-04-01

    Full Text Available Este trabajo tuvo como objetivo evaluar la técnica de inmunoelectrotransferencia (Western Blot para detectar los antígenos específicos de excreción/secreción de Hymenolepis nana en sueros de pacientes con himenolepiosis y con otras helmintiosis confirmadas. Se utilizó a Mesocricetus auratus “hamster” para obtener ejemplares adultos de H. nana. Los antígenos de excreción/secreción fueron obtenidos en el medio MEM (Minimum Essential Medium Eagle, y enfrentados con un grupo de sueros de pacientes con himenolepiosis confirmada para evaluar su calidad inmunológica y con sueros individuales de pacientes con himenolepiosis y con otras helmintiosis confirmadas para detectar mediante la técnica de “Western Blot”, los antígenos específicos de este cestode. El grupo de sueros de pacientes con himenolepiosis confirmada reconoció las bandas antigénicas de 50,1; 42,6; 38,9; 32,9; 26,3; 22,4 y 18,6 kDa; sin embargo, los sueros individuales reconocieron diferente número de bandas, siendo la de 50,1 KDa la que fue reconocida por todos ellos. Los sueros de pacientes con helmintiosis confirmadas no reconocieron la banda de 50,1 kDa; sin embargo, dieron reacción cruzada con algunas de las demás bandas, a excepción de los sueros de pacientes con cisticercosis que no reconocieron a ninguna de las bandas de estos antígenos. Se concluye que el antígeno de excreción/secreción de H. nana de 50,1 kDa es específico de este cestode por ser reconocido por todos los sueros de pacientes con himenolepiosis confirmada y no por sueros de pacientes con otras helmintiosis utilizando la técnica de “Western Blot”.

  1. Western blot data using two distinct anti-O-GlcNAc monoclonal antibodies showing unique glycosylation status on cellular proteins under 2-deoxy-d-glucose treatment

    Directory of Open Access Journals (Sweden)

    Tetsuya Okuda

    2017-02-01

    Full Text Available Protein modification by O-linked N-acetylglucosamine (O-GlcNAcylation is one of the post transcriptional modifications occurring on cellular proteins. This paper provides a data set relating to the O-GlcNAcylation of cellular proteins detected by RL2 and CTD110.6 antibodies, which are commonly used for detection of protein O-GlcNAcylation, in 2-deoxy-d-glucose (2DG-treated human teratocarcinoma NCCIT cells in support of the research article entitled “A novel, promoter-based, target-specific assay identifies 2-deoxy-d-glucose as an inhibitor of globotriaosylceramide biosynthesis” (Okuda et al., 2009 [1]. The main article described a suppressive effect of 2DG on an Sp1 target gene in NCCIT cells and discussed the relationship between the effect of 2DG and O-GlcNAcylation status of Sp1. The data in this paper complements this relationship by Western blotting and clearly showed that the 2DG treatment increased O-GlcNAcylation of cellular proteins in NCCIT cells, whereas the RL2 and CTD110.6 epitopes were detected in a different manner. The RL2 epitope was detected on Sp1 during 2DG treatment, and the level was transiently increased at 24 h. In contrast, the CTD110.6 epitope became detectable on Sp1 over 72 h after 2DG treatment, and then the other proteins containing CTD110.6 epitopes also appeared in the cell lysates and the anti-Sp1 antibody precipitates.

  2. HIV-1/2 indeterminate Western blot results: follow-up of asymptomatic blood donors in Belo Horizonte, Minas Gerais, Brazil

    Directory of Open Access Journals (Sweden)

    CARNEIRO-PROIETTI A.B.F.

    1999-01-01

    Full Text Available The clinical and public health importance of indeterminate results in HIV-1/2 testing is still difficult to evaluate in volunteer blood donors. At Fundação Hemominas, HIV-1/2 ELISA is used as the screening test and, if reactive, is followed by Western blot (WB. We have evaluated 84 blood donors who had repeatedly reactive ELISA tests for HIV-1/2, but indeterminate WB results. Sixteen of the 84 donors (19.0% had history of sexually transmitted diseases; 18/84 (21.4% informed receiving or paying for sex; 3/84 (3.6% had homosexual contact; 2/26 women (7.6% had past history of multiple illegal abortions and 3/84 (3.6% had been previously transfused. Four out of 62 donors (6.5% had positive anti-nuclear factor (Hep2, with titles up to 1:640. Parasitological examination of the stool revealed eggs of S. mansoni in 4/62 (6.4% donors and other parasites in 8/62 (12.9%. Five (5.9% of the subjects presented overt seroconversion for HIV-1/2, 43/84 (51.2% had negative results on the last visit, while 36/84 (42.9% remained WB indeterminate. Although some conditions could be found associated with the HIV-1/2 indeterminate WB results and many donors had past of risky behavior, the significance of the majority of the results remains to be determined.

  3. Discordant human T-lymphotropic virus screening with Western blot confirmation: evaluation of the dual-test algorithm for US blood donations.

    Science.gov (United States)

    Stramer, Susan L; Townsend, Rebecca L; Foster, Gregory A; Johnson, Ramona; Weixlmann, Barbara; Dodd, Roger Y

    2018-03-01

    Human T-lymphotropic virus (HTLV) blood donation screening has used a dual-testing algorithm beginning with either a chemiluminescent immunoassay or enzyme-linked immunosorbent screening assay (ELISA). Before the availability of a licensed HTLV supplemental assay, repeat-reactive (RR) samples on a first assay (Assay 1) were retested with a second screening assay (Assay 2). Donors with RR results by Assay 2 were deferred from blood donation and further tested using an unlicensed supplemental test to confirm reactivity while nonreactive (NR) donors remained eligible for donation until RR on a subsequent donation. This "dual-test" algorithm was replaced in May 2016 with the requirement that all RRs by Assay 1 be further tested by a licensed HTLV supplemental test (Western blot [WB]). In this study, we have requalified the dual-test algorithm using the available licensed HTLV WB. We tested 100 randomly selected HTLV RRs on screening Assay 1 (Abbott PRISM chemiluminescent immunoassay) but NR on screening Assay 2 (Avioq ELISA) by a Food and Drug Administration-licensed WB (MP Biomedicals) to ensure that no confirmed positives were among those that were RR by Assay 1 but NR by Assay 2. Of the 100 samples evaluated, 79 of 100 were WB seronegative, 21 of 100 indeterminate, and 0 of 100 seropositive. Of the 79 of 100 seronegative specimens, 73 of 79 did not express any bands on WB. We demonstrated that none of the 100 samples RR on Assay 1 but NR on Assay 2 were confirmed positive. This algorithm prevents such donors from requiring further testing and from being deferred. © 2018 AABB.

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

    Directory of Open Access Journals (Sweden)

    Camila Cánepa

    2015-10-01

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

  5. TLC blot (far-eastern blot) and its applications.

    Science.gov (United States)

    Taki, Takao; Gonzalez, Tania Valdes; Goto-Inoue, Naoko; Hayasaka, Takahiro; Setou, Mitsutoshi

    2009-01-01

    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, called TLC blot (far-eastern blot), is presented. Lipids separated on a HPTLC plate are blotted quantitatively. This procedure made it possible to purify individual lipids from a blotted membrane in a short time. Binding study, immunodetection, and mass spectrometric analysis are available for PVDF membrane. Furthermore, the world of molecular species imaging is opened by a scanning analysis with a combination of TLC blot and matrix-assisted laser desorption/ionization quadrupole ion trap time-of-flight mass spectrometer (TLC-Blot/MALDI-TOF MS).

  6. A comparison of the immune parameters of dogs infected with visceral leishmaniasis using Western blot and neutralization techniques Comparação dos parâmetros imunológicos de cães infectados com leishmaniose visceral usando as técnicas de Western blot e neutralização

    Directory of Open Access Journals (Sweden)

    Yeda L. Nogueira

    2007-12-01

    Full Text Available The Western blot technique was used to demonstrate the presence of antibodies in the blood of dogs that presented canine visceral leishmaniasis. This technique was used against some specific molecules present in the lysate of the promastigote form of Leshmania chagasi.Through the association of the results of the Western blot technique with the morphological alterations seen as a result of the serum neutralization technique performed in McCoy cells (which mimetizes the macrophage it was possible to observe the role of some molecules of great relevance in determining the disease in symptomatic dogs as well as that of some other molecules associated with asymptomatic infected dogs that may become transmitters as well as differentiating them as asymptomatic resistant dogs. In the sera analyses carried out during the immunobloting a variation of 9 to 27 immunoreacting bands was observed, which were then compared using Dice's similarity coefficient. In the dendrogram constructed on the basis of the coefficient, 50% similarity was observed among the total number of reagent bands with the promastigote lysate, thus creating five groups. The main difference observed related to the clinical condition of the dogs: symptomatic and asymptomatic dogs were found in separate groups. The asymptomatic group of dogs was distributed in two different places in the dendrogram because they presented two different behavior patterns regarding the cellular morphology in the serum neutralization reaction: the presence or absence of cellular lysis. According to this analysis it is possible to evaluate the immune status and associate it with specific markers observed in the reaction found in the Western blot strips.A técnica de Western blot foi utilizada para demonstrar a presença de anticorpos do soro de cães, que apresentavam leishmaniose visceral canina, contra algumas moléculas específicas no lisado da forma promastigota de Leshmania chagasi.Através da associa

  7. Delineation of pulmonary airway fluid protein fractions with HRPO binding-avidity by far-Western ligand blot and mass spectrometry analyses: a model methodology for detecting mannose-binding protein expression profiles.

    Science.gov (United States)

    Coyne, Cody P; Rashmir-Raven, Ann; Jones, Toni; Mochal, Cathleen; Linford, Robert L; Brashier, Michael; Eddy, Alison

    2009-01-01

    Limited research to date has characterized the potential for HRPO to function as a primary molecular probe. Pulmonary airway fluid was developed by non-reducing far-Western (ligand) blot analyses utilizing conjugated HRPO-strepavidin or non-conjugated HRPO without the presence of primary immunoglobulin. Endogenous esterase-like biochemical activity of fractions within pulmonary airway fluid was inactivated to determine if they were capable of biochemically converting HRPO chemiluminescent substrate. Complementary analyses modified pulmonary fluid and HRPO with beta-galactosidase and alpha-mannosidase respectively, in addition to determining the influence of mannose and maltose competitive binding on HRPO far-Western (ligand) blot analyses. Identification of pulmonary fluid fractions detected by HRPO far-Western blot analyses was determined by mass spectrometry. Modification of pulmonary fluid with beta-galactosidase, and HRPO with alpha-mannosidase in concert with maltose and mannose competitive binding analyses altered the intensity and spectrum of pulmonary fluid fractions detected by HRPO far-Western blot analysis. Identity of pulmonary airway fluid fractions detected by HRPO far-Western (ligand) blot analysis were transferrin, dynein, albumin precursor, and two 156 kDa equine peptide fragments. HRPO can function as a partially-selective primary molecular probe when applied in either a conjugated or non-conjugated form. Some protein fractions can form complexes with HRPO through molecular mechanisms that involve physical interactions at the terminal alpha-mannose-rich regions of HRPO glycan side-chains. Based on its known molecular composition and structure, HRPO provides an opportunity for the development of diagnostics methodologies relevant to disease biomarkers that possess mannose-binding avidity.

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

    Science.gov (United States)

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

    2007-01-01

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

  9. ANALYSIS OF Treponema pallidum RECOMBINANT ANTIGENS FOR DIAGNOSIS OF SYPHILIS BY WESTERN BLOTTING TECHNIQUE Análise de antígenos recombinantes de Treponema pallidum no diagnóstico da sífilis utilizando a técnica de Western Blotting

    Directory of Open Access Journals (Sweden)

    Neuza Satomi SATO

    1999-03-01

    Full Text Available Three GST fusion recombinant antigen of Treponema pallidum, described as GST-rTp47, GST-rTp17 and GST-rTp15 were analyzed by Western blotting techniques. We have tested 53 serum samples: 25 from patients at different clinical stages of syphilis, all of them presenting anti-treponemal antibody, 25 from healthy blood donors and three from patients with sexually transmitted disease (STD other than syphilis. Almost all samples from patients with syphilis presented a strong reactivity with GST-rTp17 antigen. Some samples were non-reactive or showed a weak reaction with GST-rTp47 and/or GST-rTp15, and apparently there was no correlation with the stage of disease. There was no seropositivity among blood donors. No sample reacted with purified GST. We concluded that due to their specificity these recombinant antigens can be used as GST fusion protein for development of syphilis diagnostic assays.Os antígenos recombinantes de Treponema pallidum GST-rTp47, GST-rTp17 e GST-rTp15, produzidos em fusão com glutationa S-transferase (GST em E. coli, foram analisados quanto ao potencial diagnóstico da sífilis pela técnica de Western blotting. Foram testadas 53 amostras, sendo 25 de pacientes em diferentes estágios clínicos da sífilis, com resultados positivos no teste treponêmico clássico; 25 amostras procedentes de doadores de banco de sangue, com sorologia negativa e 3 de pacientes com doença sexualmente transmissível não relacionado à sífilis. Todas as amostras de pacientes com sífilis apresentaram alta reatividade com o antígeno GST-rTp17. Quanto aos antígenos GST-rTp47 e GST-Tp15 verificou-se uma variação na presença ou na intensidade da reação em diferentes amostras de pacientes com sífilis, sem mostrar correlação com o estágio da doença. Nenhuma reatividade contra quaisquer desses antígenos foi observada com as amostras do grupo controle. Nenhuma das amostras testadas apresentaram reatividade com a GST purificada. A

  10. Use of a sensitive EnVision +-based detection system for Western blotting: avoidance of streptavidin binding to endogenous biotin and biotin-containing proteins in kidney and other tissues.

    Science.gov (United States)

    Banks, Rosamonde E; Craven, Rachel A; Harnden, Patricia A; Selby, Peter J

    2003-04-01

    Western blotting remains a central technique in confirming identities of proteins, their quantitation and analysis of various isoforms. The biotin-avidin/streptavidin system is often used as an amplification step to increase sensitivity but in some tissues such as kidney, "nonspecific" interactions may be a problem due to high levels of endogenous biotin-containing proteins. The EnVision system, developed for immunohistochemical applications, relies on binding of a polymeric conjugate consisting of up to 100 peroxidase molecules and 20 secondary antibody molecules linked directly to an activated dextran backbone, to the primary antibody. This study demonstrates that it is also a viable and sensitive alternative detection system in Western blotting applications.

  11. Introduction to protein blotting.

    Science.gov (United States)

    Kurien, Biji T; Scofield, R Hal

    2009-01-01

    Protein blotting is a powerful and important procedure for the immunodetection of proteins following electrophoresis, particularly proteins that are of low abundance. Since the inception of the protocol for protein transfer from an electrophoresed gel to a membrane in 1979, protein blotting has evolved greatly. The scientific community is now confronted with a variety of ways and means to carry out this transfer.

  12. Lectin-Array Blotting.

    Science.gov (United States)

    Pazos, Raquel; Echevarria, Juan; Hernandez, Alvaro; Reichardt, Niels-Christian

    2017-09-01

    Aberrant protein glycosylation is a hallmark of cancer, infectious diseases, and autoimmune or neurodegenerative disorders. Unlocking the potential of glycans as disease markers will require rapid and unbiased glycoproteomics methods for glycan biomarker discovery. The present method is a facile and rapid protocol for qualitative analysis of protein glycosylation in complex biological mixtures. While traditional lectin arrays only provide an average signal for the glycans in the mixture, which is usually dominated by the most abundant proteins, our method provides individual lectin binding profiles for all proteins separated in the gel electrophoresis step. Proteins do not have to be excised from the gel for subsequent analysis via the lectin array but are transferred by contact diffusion from the gel to a glass slide presenting multiple copies of printed lectin arrays. Fluorescently marked glycoproteins are trapped by the printed lectins via specific carbohydrate-lectin interactions and after a washing step their binding profile with up to 20 lectin probes is analyzed with a fluorescent scanner. The method produces the equivalent of 20 lectin blots in a single experiment, giving detailed insight into the binding epitopes present in the fractionated proteins. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

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

    Science.gov (United States)

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

    2007-12-01

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

  14. Screening for simian foamy virus infection by using a combined antigen Western blot assay: evidence for a wide distribution among Old World primates and identification of four new divergent viruses

    International Nuclear Information System (INIS)

    Hussain, Althaf I.; Shanmugam, Vedapuri; Bhullar, Vinod B.; Beer, Brigitte E.; Vallet, Dominique; Gautier-Hion, Annie; Wolfe, Nathan D.; Karesh, William B.; Kilbourn, Annelisa M.; Tooze, Zeena; Heneine, Walid; Switzer, William M.

    2003-01-01

    Simian foamy viruses (SFVs) belong to a genetically and antigenically diverse class of retroviruses that naturally infect a wide range of nonhuman primates (NHPs) and can also be transmitted to humans occupationally exposed to NHPs. Current serologic detection of SFV infection requires separate Western blot (WB) testing by using two different SFV antigens [SFV AGM (African green monkey) and SFV CPZ (chimpanzee)]. However, this method is labor intensive and validation is limited to only small numbers of NHPs. To facilitate serologic SFV testing, we developed a WB assay that combines antigens from both SFV AGM and SFV CPZ . The combined-antigen WB (CA-WB) assay was validated with 145 serum samples from 129 NHPs (32 African and Asian species) and 16 humans, all with known SFV infection status determined by PCR. Concordant CA-WB results were obtained for all 145 PCR-positive or -negative primate and human specimens, giving the assay a 100% sensitivity and specificity. In addition, no reactivity was observed in sera from persons positive for human immunodeficiency virus or human T cell lymphotropic virus (HIV/HTLV) (n = 25) or HIV/HTLV-negative U.S. blood donors (n = 100). Using the CA-WB assay, we screened 360 sera from 43 Old World primate species and found an SFV prevalence of about 68% in both African and Asian primates. We also isolated SFV from the blood of four seropositive primates (Allenopithecus nigroviridis, Trachypithecus francoisi, Hylobates pileatus, and H. leucogenys) not previously known to be infected with SFV. Phylogenetic analysis of integrase sequences from these isolates confirmed that all four SFVs represent new, distinct, and highly divergent lineages. These results demonstrate the ability of the CA-WB assay to detect infection in a large number of NHP species, including previously uncharacterized infections with divergent SFVs

  15. Detection of Babesia and Theileria species infection in cattle from Portugal using a reverse line blotting method.

    Science.gov (United States)

    Silva, M G; Marques, P X; Oliva, A

    2010-12-15

    Babesiosis and Theileriosis are tick-borne diseases widespread in tropical and sub-tropical regions with high economic impact worldwide. In Portugal there are at least 4 tick vectors known to be competent for the transmission of Babesia and Theileria sp. identified: Rhipicephalus bursa, Rhipicephalus (Boophilus) annulatus, Ixodes ricinus and Haemaphysalis punctata. All these potential Babesia and Theileria tick vectors are widely distributed in Portugal, although they are predominant in the Southern region. In this study, 1104 cattle blood samples were randomly collected from Central and Southern regions of Portugal and analyzed by PCR-reverse line blotting (RLB) for the detection of Babesia and Theileria sp. Testing indicated that 74.7% of the bovines tested were positive for either Babesia and/or Theileria sp. In addition, five different apicomplexan species, namely, Theileria buffeli, Theileria annulata, Babesia divergens, Babesia bovis, and Babesia bigemina were detected by RLB among the bovines tested. T. buffeli was the most frequently found species, being present in 69.9% of the positive samples either as single infections (52.4%), or as mixed infections (17.5%). The Babesia specie most frequently found was B. divergens, detected in 4.2% of the infected bovines. Overall, infected bovines were found in all regions tested; however the highest number of infected bovines was observed in Évora district (96.2%) and in cattle from Limousin breeds (81.7%). The results indicate widespread Babesia and Theileria infections in Portuguese bovines, suggesting the need for improved control of ticks and tick-borne diseases. Copyright © 2010 Elsevier B.V. All rights reserved.

  16. Northern blotting analysis

    DEFF Research Database (Denmark)

    Josefsen, Knud; Nielsen, Henrik

    2011-01-01

    is analysed by hybridization to one or more specific probes that are labelled for subsequent detection. Northern blotting is relatively simple to perform, inexpensive, and not plagued by artefacts. Recent developments of hybridization membranes and buffers have resulted in increased sensitivity closing...

  17. Detection of KatG Gen Mutation on Mycobacterium Tuberculosis by Means of PCR-Dot Blot Hybridization with 32P Labeled Oligonucleotide Probe Methods

    International Nuclear Information System (INIS)

    Maria Lina R; Budiman Bela; Andi Yasmon

    2009-01-01

    Handling and controlling of tuberculosis, a disease caused by Mycobacterium tuberculosis (MTB), is now complicated since there are many MTBs that are resistant against anti-tuberculosis drugs such as isoniazid. The drug resistance could occurred due to the inadequate and un-regular drug utilization that cause gene mutation of the drug target such as katG gene for isoniazid. The molecular biology techniques such as the PCR- dot blot hybridization with radioisotope ( 32 P) labeled oligonucleotide probe, has been reported as a technique that is more sensitive and rapid for detection of gene mutations related with drug resistances. Hence, the aim of this study was to apply the PCR- dot blot hybridization technique using 32 P labeled oligonucleotide probe for detection of single mutation at codon 315 of katG gene of MTBs that rise the isoniazid resistance. In this study, we used 89 sputum specimens and a standard MTB (MTB H 37 RV) as a control. DNA extractions were performed by the BOOM method and the phenol chloroform for sputum samples and standard MTB, respectively. Primers used for PCR technique were Pt8 and Pt9 and RTB59 and RTB36 for detecting tuberculosis causing Mycobacterium and the existence of katG gene, respectively. Both of the primers are specific for IS6110 region and katG gene, respectively. PCR products were detected by an agarose gel electrophoresis technique. Dot blot hybridization with 32 P-oligonucleotide probe 315mu was performed to detect mutation at codon 315 of tested samples. Results of the PCR using primer Pt8 and Pt9 showed that all sputum specimens had positive results. Mutation detection by PCR- dot blot hybridization with 32 P-oligonucleotide probe 315mu, revealed that 11 of 89 tested samples had a mutation at their codon 315 of katG gene. Based upon these results, it is concluded that PCR-dot blot hybridization with 32 P-oligonucleotide probe is a technique that is rapid and highly specific and sensitive for detection of mutation at codon

  18. A Western-blot assay for the detection of antibodies against pathogenic Leptospira serogroups with recombinant outer membrane protein LipL32

    OpenAIRE

    Hong-yuan DUAN; Zhi-guo LIU; Shao-fu QIU; Bin HE; Hai ZHAO; Li-hua SONG; Hong ZHU; Qing DUAN

    2011-01-01

    Objective To provide a possible antigen for rapid serodiagnosis of leptospirosis,the present study focused on the activity of immune-reaction and cross-reaction between outer membrane protein LipL32 and multi-serogroup anti-pathogenic Leptospira antibodies.Methods Based on the given sequence of LipL32 gene of Leptospira icterohaemorrhagiae strain 56601,the primer pair was designed and the DNA fragment was amplified by PCR.The amplified product was inserted into vector pET-28a-(c) to construct...

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

    DEFF Research Database (Denmark)

    Vigelsø Hansen, Andreas; Dybboe, Rie; Hansen, Christina Neigaard

    2015-01-01

    SF and RP was measured in relation to ageing, muscle atrophy, and different muscle fiber type composition, respectively. A stronger linearity of SF and β-actin compared with GAPDH and α-tubulin was observed. The methodological variation was relatively low in all four methods (4-11%). Protein level...... [β-actin, glyceraldehyde 3-phosphate dehydrogenase (GAPDH), and α-tubulin], as well as TP loaded measured by Stain-Free technology (SF) as normalization tool were tested. This was done using skeletal muscle samples from men subjected to physiological conditions often investigated in applied...... physiology where the intervention has been suggested to impede normalization (ageing, muscle atrophy, and different muscle fiber type composition). The linearity of signal and the methodological variation coefficient was obtained. Furthermore, the inter- and intraindividual variation in signals obtained from...

  20. Immunocytochemical electron microscopic study and western blot analysis of paramyosin in different invertebrate muscle cell types of the fruit fly Drosophila melanogaster, the earthworm Eisenia foetida, and the snail Helix aspersa.

    Science.gov (United States)

    Royuela, M; García-Anchuelo, R; Arenas, M I; Cervera, M; Fraile, B; Paniagua, R

    1996-04-01

    The presence and distribution pattern of paramyosin have been examined in different invertebrate muscle cell types by means of Western blot analysis and electron microscopy immunogold labelling. The muscles studied were: transversely striated muscle with continuous Z lines (flight muscle from Drosophila melanogaster), transversely striated muscle with discontinuous Z lines (heart muscle from the snail Helix aspersa), obliquely striated body wall muscle from the earthworm Eisenia foetida, and smooth muscles (retractor muscle from the snail and pseudoheart outer muscular layer from the earthworm). Paramyosin-like immunoreactivity was localized in thick filaments of all muscles studied. Immunogold particle density was similar along the whole thick filament length in insect flight muscle but it predominated in filament tips of fusiform thick filaments in both snail heart and earthworm body wall musculature when these filaments were observed in longitudinal sections. In obliquely sectioned thick filaments, immunolabelling was more abundant at the sites where filaments disappeared from the section. These results agree with the notion that paramyosin extended along the whole filament length, but that it can only be immunolabelled when it is not covered by myosin. In all muscles examined, immunolabelling density was lower in cross-sectioned myofilaments than in longitudinally sectioned myofilaments. This suggests that paramyosin does not form a continuous filament. The results of a semiquantitative analysis of paramyosin-like immunoreactivity indicated that it was more abundant in striated than in smooth muscles, and that, within striated muscles, transversely striated muscles contain more paramyosin than obliquely striated muscles.

  1. Approaches for Reverse Line Blot-Based Detection of Microbial Pathogens in Ixodes ricinus Ticks Collected in Austria and Impact of the Chosen Method.

    Science.gov (United States)

    Schötta, Anna-Margarita; Wijnveld, Michiel; Stockinger, Hannes; Stanek, Gerold

    2017-07-01

    Ticks transmit a large number of pathogens capable of causing human disease. In this study, the PCR-reverse line blot (RLB) method was used to screen for pathogens in a total of 554 Ixodes ricinus ticks collected from all provinces of Austria. These pathogens belong to the genera Borrelia , Rickettsiae , Anaplasma / Ehrlichia (including " Candidatus Neoehrlichia"), Babesia , and Coxiella The pathogens with the highest detected prevalence were spirochetes of the Borrelia burgdorferi sensu lato complex, in 142 ticks (25.6%). Borrelia afzelii (80/142) was the most frequently detected species, followed by Borrelia burgdorferi sensu stricto (38/142) and Borrelia valaisiana (36/142). Borrelia garinii/Borrelia bavariensis , Borrelia lusitaniae , and Borrelia spielmanii were found in 28 ticks, 5 ticks, and 1 tick, respectively. Rickettsia spp. were detected in 93 ticks (16.8%): R. helvetica (39/93), R. raoultii (38/93), R. monacensis (2/93), and R. slovaca (1/93). Thirteen Rickettsia samples remain uncharacterized. " Candidatus Neoehrlichia mikurensis," Babesia spp. ( B. venatorum , B. divergens , B. microti ), and Anaplasma phagocytophilum were found in 4.5%, 2.7%, and 0.7%, respectively. Coxiella burnetii was not detected. Multiple microorganisms were detected in 40 ticks (7.2%), and the cooccurrence of Babesia spp. and " Candidatus Neoehrlichia mikurensis" showed a significant positive correlation. We also compared different PCR-RLBs for detection of Borrelia burgdorferi sensu lato and Rickettsia spp. and showed that different detection approaches provide highly diverse results, indicating that analysis of environmental samples remains challenging. IMPORTANCE This study determined the wide spectrum of tick-borne bacterial and protozoal pathogens that can be encountered in Austria. Surveillance of (putative) pathogenic microorganisms occurring in the environment is of medical importance, especially when those agents can be transmitted by ticks and cause disease. The

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

    Science.gov (United States)

    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.

  3. A combined RT-PCR and dot-blot hybridization method reveals the coexistence of SJNNV and RGNNV betanodavirus genotypes in wild meagre (Argyrosomus regius).

    Science.gov (United States)

    Lopez-Jimena, B; Cherif, N; Garcia-Rosado, E; Infante, C; Cano, I; Castro, D; Hammami, S; Borrego, J J; Alonso, M C

    2010-10-01

    To detect the possible coexistence of striped jack nervous necrosis virus (SJNNV) and red-spotted grouper nervous necrosis virus (RGNNV) genotypes in a single fish, a methodology based on the combination of PCR amplification and blot hybridization has been developed and applied in this study. The degenerate primers designed for the PCR procedure target the T4 region within the capsid gene, resulting in the amplification of both genotypes. The subsequent hybridization of these amplification products with two different specific digoxigenin-labelled probes resulted in the identification of both genotypes separately. The application of the RT-PCR protocol to analyse blood samples from asymptomatic wild meagre (Argyrosomus regius) specimens has shown a 46.87% of viral nervous necrosis virus carriers. The combination of RT-PCR and blot hybridization increases the detection rate up to 90.62%, and, in addition, it has shown the coexistence of both genotypes in 18 out of the 32 specimens analysed (56.25%). This study reports the coexistence of betanodaviruses belonging to two different genotypes (SJNNV and RGNNV) in wild fish specimens. This is the first report demonstrating the presence of SJNNV and RGNNV genotypes in the same specimen. This study also demonstrates a carrier state in this fish species for the first time. © 2010 The Authors. Journal compilation © 2010 The Society for Applied Microbiology.

  4. Adapting Western research methods to indigenous ways of knowing.

    Science.gov (United States)

    Simonds, Vanessa W; Christopher, Suzanne

    2013-12-01

    Indigenous communities have long experienced exploitation by researchers and increasingly require participatory and decolonizing research processes. We present a case study of an intervention research project to exemplify a clash between Western research methodologies and Indigenous methodologies and how we attempted reconciliation. We then provide implications for future research based on lessons learned from Native American community partners who voiced concern over methods of Western deductive qualitative analysis. Decolonizing research requires constant reflective attention and action, and there is an absence of published guidance for this process. Continued exploration is needed for implementing Indigenous methods alone or in conjunction with appropriate Western methods when conducting research in Indigenous communities. Currently, examples of Indigenous methods and theories are not widely available in academic texts or published articles, and are often not perceived as valid.

  5. Automated design of genomic Southern blot probes

    Directory of Open Access Journals (Sweden)

    Komiyama Noboru H

    2010-01-01

    Full Text Available Abstract Background Sothern blotting is a DNA analysis technique that has found widespread application in molecular biology. It has been used for gene discovery and mapping and has diagnostic and forensic applications, including mutation detection in patient samples and DNA fingerprinting in criminal investigations. Southern blotting has been employed as the definitive method for detecting transgene integration, and successful homologous recombination in gene targeting experiments. The technique employs a labeled DNA probe to detect a specific DNA sequence in a complex DNA sample that has been separated by restriction-digest and gel electrophoresis. Critically for the technique to succeed the probe must be unique to the target locus so as not to cross-hybridize to other endogenous DNA within the sample. Investigators routinely employ a manual approach to probe design. A genome browser is used to extract DNA sequence from the locus of interest, which is searched against the target genome using a BLAST-like tool. Ideally a single perfect match is obtained to the target, with little cross-reactivity caused by homologous DNA sequence present in the genome and/or repetitive and low-complexity elements in the candidate probe. This is a labor intensive process often requiring several attempts to find a suitable probe for laboratory testing. Results We have written an informatic pipeline to automatically design genomic Sothern blot probes that specifically attempts to optimize the resultant probe, employing a brute-force strategy of generating many candidate probes of acceptable length in the user-specified design window, searching all against the target genome, then scoring and ranking the candidates by uniqueness and repetitive DNA element content. Using these in silico measures we can automatically design probes that we predict to perform as well, or better, than our previous manual designs, while considerably reducing design time. We went on to

  6. The origins of Western mind-body exercise methods.

    Science.gov (United States)

    Hoffman, Jonathan; Gabel, C Philip

    2015-11-02

    Background: Over recent decades, mind-body exercise methods have gained international popularity and importance in the management of musculoskeletal disorders. Objectives: The scope of this paper was to investigate: the origins of Western mind-body methods, their philosophies, exercises, and relationship with mainstream healthcare over the last two centuries. Major findings: Within a few decades of the turn of the 20th century, a cluster of mind-body exercise methods emerged from at least six pioneering founders: Checkley, Müller, Alexander, Randell, Pilates, and Morris. Each was based upon a similar exercise philosophy and similar functional movement-harmonizing exercises. This renaissance of independent mind-body schools occurred in parallel with the demise of the 18th and 19th century gymnasium Physical Culture movement and the concurrent emergence of bodybuilding and strength training. Even though mostly forgotten today, Western mind-body exercise methods enjoyed celebrated success during the first half of the 20th century, were hailed by medical and allied health practitioners and practiced by millions from society's elite to deprived minorities. Conclusions: Rediscovering the Western mind-body exercise movement is hoped to facilitate official healthcare establishment recognition of this kind of training as an integral entity. This may widen research opportunities and consolidate approaches toward: optimal musculoskeletal rehabilitation and injury prevention, promotion of a healthy active lifestyle environment in the modern world, and enhancement of the natural pain-free human athletic look, feel, and performance.

  7. Human Immunodeficiency Virus (HIV types Western blot (WB band profiles as potential surrogate markers of HIV disease progression and predictors of vertical transmission in a cohort of infected but antiretroviral therapy naïve pregnant women in Harare, Zimbabwe

    Directory of Open Access Journals (Sweden)

    Chirenje Mike Z

    2011-01-01

    Full Text Available Abstract Background Expensive CD4 count and viral load tests have failed the intended objective of enabling access to HIV therapy in poor resource settings. It is imperative to develop simple, affordable and non-subjective disease monitoring tools to complement clinical staging efforts of inexperienced health personnel currently manning most healthcare centres because of brain drain. Besides accurately predicting HIV infection, sequential appearance of specific bands of WB test offers a window of opportunity to develop a less subjective tool for monitoring disease progression. Methods HIV type characterization was done in a cohort of infected pregnant women at 36 gestational weeks using WB test. Student-t test was used to determine maternal differences in mean full blood counts and viral load of mothers with and those without HIV gag antigen bands. Pearson Chi-square test was used to assess differences in lack of bands appearance with vertical transmission and lymphadenopathy. Results Among the 64 HIV infected pregnant women, 98.4% had pure HIV-1 infection and one woman (1.7% had dual HIV-1/HIV-2 infections. Absence of HIV pol antigen bands was associated with acute infection, p = 0.002. All women with chronic HIV-1 infection had antibody reactivity to both the HIV-1 envelope and polymerase antigens. However, antibody reactivity to gag antigens varied among the women, being 100%, 90%, 70% and 63% for p24, p17, p39 and p55, respectively. Lack of antibody reactivity to gag p39 antigen was associated with disease progression as confirmed by the presence of lymphadenopathy, anemia, higher viral load, p = 0.010, 0.025 and 0.016, respectively. Although not statistically significant, women with p39 band missing were 1.4 times more likely to transmit HIV-1 to their infants. Conclusion Absence of antibody reactivity to pol and gag p39 antigens was associated with acute infection and disease progression, respectively. Apart from its use in HIV disease

  8. Post-staining electroblotting for efficient and reliable peptide blotting.

    Science.gov (United States)

    Lee, Der-Yen; Chang, Geen-Dong

    2015-01-01

    Post-staining electroblotting has been previously described to transfer Coomassie blue-stained proteins from polyacrylamide gel onto polyvinylidene difluoride (PVDF) membranes. Actually, stained peptides can also be efficiently and reliably transferred. Because of selective staining procedures for peptides and increased retention of stained peptides on the membrane, even peptides with molecular masses less than 2 kDa such as bacitracin and granuliberin R are transferred with satisfactory results. For comparison, post-staining electroblotting is about 16-fold more sensitive than the conventional electroblotting for visualization of insulin on the membrane. Therefore, the peptide blots become practicable and more accessible to further applications, e.g., blot overlay detection or immunoblotting analysis. In addition, the efficiency of peptide transfer is favorable for N-terminal sequence analysis. With this method, peptide blotting can be normalized for further analysis such as blot overlay assay, immunoblotting, and N-terminal sequencing for identification of peptide in crude or partially purified samples.

  9. Social Capital in Western and Eastern Europe: Method of Measurement

    DEFF Research Database (Denmark)

    Hjøllund, L.; Svendsen, Gert Tinggaard

    2000-01-01

    The purpose of this paper is to suggest a standard method of measurement for social capital. Various authors have investigated the influence of social capital on economic growth but still social capital has not been measured in any satisfactory way. So far, each survey has used its own ad hoc met...

  10. [The methods of Western medicine in on ancient medicine].

    Science.gov (United States)

    Ban, Deokjin

    2010-06-30

    The treatise On Ancient Medicine attests that questions of method were being debated both in medicine and in philosophy and is important evidence of cross-discipline methodological controversy. The treatise On Ancient Medicine is the first attempt in the history of Greek thought to provide a detailed account of the development of a science from a starting point in observation and experience. The author of it criticizes philosophical physicians who attempt to systematized medicine by reducing it to the interaction of one or more of the opposites hot, cold, wet, and dry, factors. He regards the theory of his opponents as hypothesis(hypothesis). Medicine has long been in possession of both an archē and a hodos, a principle and a method, which have enabled it to make discoveries over a long period of time. As far as method is concerned, the traditional science of medicine attained the knowledge of the visible by starting from observation and experience, but it recommended the use of reasoning and analogies with familiar objects as a means of learning about the invisible. It also utilized inference from the visible to the visible(epilogismos) and inference from the visible to the invisible(analogismos). The use of analogy as a means of learning about the obscure was also part of the common heritage of early philosophy and medicine. But the author's use of the analogical method distinguishes it from Empedocles' well-known analogy comparisons of the eye to a lantern and the process of respiration to the operations of a clepsydra. According to the author, traditional science of medicine used functional analogy like wine example and cheese example to know the function of humors within the body and utilized structured analogy like a tube example and a cupping instrument example to acknowledge an organ or structure within the body. But the author didn't distinguish between the claim that medicine has a systematic method of making discoveries and very different claim that it

  11. Zinc blotting assay for detection of zinc binding prolamin in barley (Hordeum vulgare) grain

    DEFF Research Database (Denmark)

    Uddin, Mohammad Nasir; Nielsen, Ane Langkilde-Lauesen; Vincze, Eva

    2014-01-01

    In plants, zinc is commonly found bound to proteins. In barley (Hordeum vulgare), major storage proteins are alcohol-soluble prolamins known as hordeins, and some of them have the potential to bind or store zinc. 65Zn overlay and blotting techniques have been widely used for detecting zinc......-binding protein. However, to our knowledge so far this zinc blotting assay has never been applied to detect a prolamin fraction in barley grains. A radioactive zinc (65ZnCl2) blotting technique was optimized to detect zinc-binding prolamins, followed by development of an easy-to-follow nonradioactive colorimetric...... zinc blotting method with a zinc-sensing dye, dithizone. Hordeins were extracted from mature barley grain, separated by SDS-PAGE, blotted on a membrane, renatured, overlaid, and probed with zinc; subsequently, zinc-binding specificity of certain proteins was detected either by autoradiography or color...

  12. [Efficacy on chronic obstructive pulmonary disease at stable stage treated with cutting method and western medication].

    Science.gov (United States)

    Xu, Jian-hua; Xu, Bin; Deng, Yan-qing

    2014-10-01

    To compare the difference in clinical efficacy on chronic obstructive pulmonary disease (COPD) at stable stage in the patients among the combined therapy of cutting method and western medication (combined therapy), simple cutting method and simple western medication. One hundred and twenty cases of COPD were randomized into three groups, 40 cases in each one. In the cutting method group, for excessive phlegm pattern/syndrome, Feishu (BL 13), Danzhong (CV 17), Dingchuan (EX-B 1) and Yuji (LU 10) were selected as the main acupoints, and Lieque (LU 7) and Pianli (LI 6) were as the supplementary acupoints. For the pattern/syndrome of failure to consolidate kidney primary, Shenshu (BL 23), Pishu (BL 20), Guanyuan (CV 4) and Yuji (LU 10) were selected as main acupoints, and Jueyinshu (BL 14) and Zusanli (ST 36) were as the supplementary acupoint. Three acupoints were selected alternatively in each treatment and the cutting method was applied once every 10 days. Three treatments made one session. Two sessions of treatment were required. In the western medication group, salbutamol sulfate aerosol, one press (200 μg/press) was used each night, as well as salmeterol xinafoate and fluticasone propionate powder for inhalation, one inhalation each night. The treatment of 1 month made one session. Two sessions were required. In the combined therapy group, the cutting method and western medication were applied in combination. The results of clinical symptom score, lung function test, arterial blood gas analysis, degree of inflation as well as clinical efficacy were observed before and after treatment in each group. Except the degree of lung inflation, the clinical symptom score, indices of lung function test, partial pressure of arterial blood gas (PaO2) and partial pressure of carbon dioxide (PaCO2) were all obviously improved after treatment as compared with those before treatment in each group (all Psyndrome differentiation and the combined therapy with western medication

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

    Science.gov (United States)

    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. © 2016 Wiley Periodicals, Inc.

  14. Evaluation of five methods for total DNA extraction from western corn rootworm beetles.

    Directory of Open Access Journals (Sweden)

    Hong Chen

    Full Text Available BACKGROUND: DNA extraction is a routine step in many insect molecular studies. A variety of methods have been used to isolate DNA molecules from insects, and many commercial kits are available. Extraction methods need to be evaluated for their efficiency, cost, and side effects such as DNA degradation during extraction. METHODOLOGY/PRINCIPAL FINDINGS: From individual western corn rootworm beetles, Diabrotica virgifera virgifera, DNA extractions by the SDS method, CTAB method, DNAzol reagent, Puregene solutions and DNeasy column were compared in terms of DNA quantity and quality, cost of materials, and time consumed. Although all five methods resulted in acceptable DNA concentrations and absorbance ratios, the SDS and CTAB methods resulted in higher DNA yield (ng DNA vs. mg tissue at much lower cost and less degradation as revealed on agarose gels. The DNeasy kit was most time-efficient but was the costliest among the methods tested. The effects of ethanol volume, temperature and incubation time on precipitation of DNA were also investigated. The DNA samples obtained by the five methods were tested in PCR for six microsatellites located in various positions of the beetle's genome, and all samples showed successful amplifications. CONCLUSION/SIGNIFICANCE: These evaluations provide a guide for choosing methods of DNA extraction from western corn rootworm beetles based on expected DNA yield and quality, extraction time, cost, and waste control. The extraction conditions for this mid-size insect were optimized. The DNA extracted by the five methods was suitable for further molecular applications such as PCR and sequencing by synthesis.

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

    International Nuclear Information System (INIS)

    Bodkin, D.K.

    1985-01-01

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

  16. Utility of qualitative methods in a clinical setting: perinatal care in the Western Province.

    Science.gov (United States)

    Jayasuriya, V

    2012-03-01

    A peculiar paradox that has been observed in previous studies of antenatal care is where patients are satisfied with the services despite obvious lack of basic facilities. Qualitative methods were used to describe the experience of perinatal care in the Western province with the objective of demonstrating application of this method in a clinical setting. This paper used a 'naturalistic' approach of qualitative methods. In-depth interviews conducted with 20 postnatal mothers delivering in tertiary care institutions in the Western province was tape recorded, transcribed and content analysed. To ensure objectivity and validity of results, the principle investigator received only the anonymised data to prevent any prejudices or pre-conceptions affecting the results. The main themes emerging from the text demonstrated 'naïve trust' in the carer and a state of 'hero worship' where patients were distanced and therefore unable and unwilling to query the decisions made by the carers. This is similar to a state of patient-carer relationship described in a published model known as guarded alliance, where the relationship develops though four phases based on the level of trust and confidence in the relationship. This state explains not only why patients fail to recognise and report any deficiencies in the services but also the need for them to justify the behaviour of caregivers even when it amounts to incompetence and negligence. Qualitative methods allow the researcher to capture experiences in its 'natural' form rather than based on pre-determined protocols or plans, which may be limited to our own understanding and expectations and therefore unable to explain many idiosyncrasies of the programmes. This paper argues favourably for the use of qualitative methods in other clinical settings.

  17. Evaluation of two methods of estimating larval habitat productivity in western Kenya highlands

    Directory of Open Access Journals (Sweden)

    Munga Stephen

    2011-06-01

    Full Text Available Abstract Background Malaria vector intervention and control programs require reliable and accurate information about vector abundance and their seasonal distribution. The availability of reliable information on the spatial and temporal productivity of larval vector habitats can improve targeting of larval control interventions and our understanding of local malaria transmission and epidemics. The main objective of this study was to evaluate two methods of estimating larval habitat productivity in the western Kenyan highlands, the aerial sampler and the emergence trap. Methods The study was conducted during the dry and rainy seasons in 2008, 2009 and 2010. Aerial samplers and emergence traps were set up for sixty days in each season in three habitat types: drainage ditches, natural swamps, and abandoned goldmines. Aerial samplers and emergence traps were set up in eleven places in each habitat type. The success of each in estimating habitat productivity was assessed according to method, habitat type, and season. The effect of other factors including algae cover, grass cover, habitat depth and width, and habitat water volume on species productivity was analysed using stepwise logistic regression Results Habitat productivity estimates obtained by the two sampling methods differed significantly for all species except for An. implexus. For for An. gambiae s.l. and An. funestus, aerial samplers performed better, 21.5 and 14.6 folds, than emergence trap respectively, while the emergence trap was shown to be more efficient for culicine species. Seasonality had a significant influence on the productivity of all species monitored. Dry season was most productive season. Overall, drainage ditches had significantly higher productivity in all seasons compared to other habitat types. Algae cover, debris, chlorophyll-a, and habitat depth and size had significant influence with respect to species. Conclusion These findings suggest that the aerial sampler is the

  18. Blotting from PhastGel to Membranes by Ultrasound.

    Science.gov (United States)

    Kost, Joseph; Azagury, Aharon

    2015-01-01

    Ultrasound based approach for enhanced protein blotting is proposed. Three minutes of ultrasound exposure (1 MHz, 2.5 W/cm(2)) was sufficient for a clear transfer of proteins from a polyacrylamide gel (PhastGel) to nitrocellulose or Nylon 66 Biotrans membrane. The proteins evaluated were prestained sodium dodecyl sulfate-polyacrylamide standards (18,500-106,000 Da) and 14C-labeled Rainbow protein molecular weight markers (14,300-200,000 Da).

  19. Effects of the control method (Goč variety in selection forest management in Western Serbia

    Directory of Open Access Journals (Sweden)

    Medarević M.

    2010-01-01

    Full Text Available The control method, one of the most reliable methods of selection forest management, has been applied in selection forests of western Serbia in a somewhat modified form (Goč variety for fifty years. This paper analyzes the effects of the control method, i.e. its Goč variety, in the period from 1960/70 - 2000. It is based on the data of five successive complete inventories of the Forest Management Unit (FMU 'Tara', whose high selection forest of spruce, fir and beech (Piceo-Abieti-Fagetum subass. typicum trees on diluvium, brown and illimerised soil on limestone, and on limestone in formation with hornfels, are the best quality and the most spacious forests in the Management Class MC 491/1. The effects were monitored through the changes in the distribution of the number of trees and volume per diameter classes, separately for fir as the protagonist of the selection structure, and collectively at the level of a compartment, a typical representative of MC 491/1. Also, the analysis included the changes in the number of trees, volume, current volume increment, yield, and number of recruited trees per unit area (1 ha by tree species in MC 491/1, occupying an area of 2,648.78 ha. The study results show that in the study period the average volume in MC 491/1 increased by 18.8%, the percentage of conifers increased from 66.0% to 78.5%, and the bearer of the changes was fir. The volume of the mean fir tree increased by 35.9% and it attained 1.086 m3. The volume increment increased by 15.7%. The selection structure of conifers was satisfactory, but there were problems with beech regeneration, in its stable presence and in its achievement of the targeted structure. The number of trees per unit area (1 ha decreased, which in the long run could have detrimental consequences, but the sustainability in general was satisfactory. The levels of regeneration and recruitment were satisfactory. The health of the trees was improved; the stands were healthy, vital

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

    Science.gov (United States)

    1992-09-16

    William Lafferty, Departments of Medicine. Laboratori’ Medicine. Epidenrioloý k. .nd Thomas S. Inui, Pamela H. Louie, Carol A. Gates, Biostatistics ...and dentistry . Finally, the psychosoclal onsequences of lWB are considerable.16 1655;LBISOTO2 JAB 13:33 03-17-92 P0003 U3 C9439 0021 11655 JGLM Nov

  1. The effect of interpolation methods in temperature and salinity trends in the Western Mediterranean

    Directory of Open Access Journals (Sweden)

    M. VARGAS-YANEZ

    2012-04-01

    Full Text Available Temperature and salinity data in the historical record are scarce and unevenly distributed in space and time and the estimation of linear trends is sensitive to different factors. In the case of the Western Mediterranean, previous works have studied the sensitivity of these trends to the use of bathythermograph data, the averaging methods or the way in which gaps in time series are dealt with. In this work, a new factor is analysed: the effect of data interpolation. Temperature and salinity time series are generated averaging existing data over certain geographical areas and also by means of interpolation. Linear trends from both types of time series are compared. There are some differences between both estimations for some layers and geographical areas, while in other cases the results are consistent. Those results which do not depend on the use of interpolated or non-interpolated data, neither are influenced by data analysis methods can be considered as robust ones. Those results influenced by the interpolation process or the factors analysed in previous sensitivity tests are not considered as robust results.

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

    Science.gov (United States)

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

    2011-12-01

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

  3. Aquifer Characterization from Surface Geo-electrical Method, western coast of Maharashtra, India

    Science.gov (United States)

    DAS, A.; Maiti, D. S.

    2017-12-01

    assess and manage properly in western Maharashtra. The current method which include DC resistivity inversion could be applicable further in hydrological characterization in tangled coastal parts of India.

  4. Methods for monitoring land subsidence and earth fissures in the Western USA

    Directory of Open Access Journals (Sweden)

    K. C. Fergason

    2015-11-01

    Full Text Available Depletion of groundwater resources in many deep alluvial basin aquifers in the Western USA is causing land subsidence, as it does in many regions worldwide. Land subsidence can severely and adversely impact infrastructure by changing the ground elevation, ground slope (grade and through the development of ground cracks known as earth fissures that can erode into large gullies. Earth fissures have the potential to compromise the foundations of dams, levees, and other infrastructure and cause failure. Subsequent to an evaluation of the overall subsidence experienced in the vicinity of subsidence-impacted infrastructure, a detailed investigation to search for earth fissures, and design and/or mitigation of potentially effected infrastructure, a focused monitoring system should be designed and implemented. Its purpose is to provide data, and ultimately knowledge, to reduce the potential adverse impacts of land subsidence and earth fissure development to the pertinent infrastructure. This risk reduction is realized by quantifying the rate and distribution of ground deformation, and to detect ground rupture if it occurs, in the vicinity of the infrastructure. The authors have successfully designed and implemented monitoring systems capable of quantifying rates and distributions of ground subsidence and detection of ground rupture at multiple locations throughout the Western USA for several types of infrastructure including dams, levees, channels, basins, roadways, and mining facilities. Effective subsidence and earth fissure monitoring requires understanding and quantification of historic subsidence, estimation of potential future subsidence, delineation of the risk for earth fissures that could impact infrastructure, and motivation and resources to continue monitoring through time. A successful monitoring system provides the means to measure ground deformation, grade changes, displacement, and anticipate and assess the potential for earth fissuring

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

    International Nuclear Information System (INIS)

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

    1986-01-01

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

  6. Can Western quality improvement methods transform the Russian health care system?

    Science.gov (United States)

    Tillinghast, S J

    1998-05-01

    The Russian health care system largely remains the same system that was in place during the existence of the Soviet Union. It is almost entirely state owned and operated, although ownership and management have developed from the central government to the oblast (province). The ZdravReform (Health Reform) Program (ZRP) in Russia, which began in 1993, included the goal of improving the quality and cost-effectiveness of the health care system. Work on introducing continuous quality improvement (CQI), evidence-based practice guidelines, and indicators of quality was conducted in 1995-1996. INTRODUCING EVIDENCE-BASED MEDICINE: As a result of the poor quality of Russian-language medical journals and the inability to gain access to the knowledge available in Western medical literature, Russian medical practices have not kept up with the rapid evolution of evidence-based medical practice that has begun transforming Western medicine. A number of evidence-based clinical practice guidelines were translated and disseminated to Russian-speaking physicians working in facilities participating in ZRP in Russia and Central Asia. Given the limitations of existing measures of the quality of care, indicators were developed for participating ambulatory polyclinics in several oblasts in Siberia. Russian physicians responsible for quality of care for their respective oblasts formed a working group to develop the indicators. A clinical information system that would provide automated collection and analysis of the indicator data-as well as additional patient record information-was also developed. CQI activities, entailing a multidisciplinary, participatory team approach, were conducted in four oblasts in western Siberia. Projects addressed the management of community-acquired pneumonia and reduction of length of stay after myocardial infarction (MI). One of the oblasts provided an example of a home-grown evidence-based protocol for post-MI care, which was adopted in the other three oblasts

  7. Silver and gold nanoparticle coated membranes applied to protein dot blots

    International Nuclear Information System (INIS)

    Xie, F.; Drozdowicz-Tomsia, K.; Shtoyko, T.; Goldys, E. M.

    2011-01-01

    Detection and identification of low abundance biomarker proteins is frequently based on various types of membrane-based devices. Lowering of the protein detection limits is vital in commercial applications such as lateral flow assays and in Western blots widely used in proteomics. These currently suffer from insufficient detection sensitivity and low retention for small 2–5 kDa proteins. In this study, we report the deposition of two types of metal nanoparticles: gold colloids (50–95 nm diameter) and silver fractals onto a range of commonly used types of membranes including polyvinylidene fluoride (PVDF). Due to strong affinity of proteins to noble metals, such modified membranes have the potential to effectively capture trace proteins preventing their loss. The membranes modified by metal particles were characterized optically and by SEM. The membrane performance in protein dot blots was evaluated using the protein—fluorophore conjugates Deep Purple-bovine serum albumin and fluorescein—human serum albumin. We found that the metal nanoparticles increase light extinction by metals, which is balanced by increased fluorescence, so that the effective fluorescence signal is unchanged. This feature combined with the capture of proteins by the nanoparticles embedded in the membrane increases the detection limit of membrane assays.

  8. Far Western: probing membranes.

    Science.gov (United States)

    Einarson, Margret B; Pugacheva, Elena N; Orlinick, Jason R

    2007-08-01

    INTRODUCTIONThe far-Western technique described in this protocol is fundamentally similar to Western blotting. In Western blots, an antibody is used to detect a query protein on a membrane. In contrast, in a far-Western blot (also known as an overlay assay) the antibody is replaced by a recombinant GST fusion protein (produced and purified from bacteria), and the assay detects the interaction of this protein with target proteins on a membrane. The membranes are washed and blocked, incubated with probe protein, washed again, and subjected to autoradiography. The GST fusion (probe) proteins are often labeled with (32)P; alternatively, the membrane can be probed with unlabeled GST fusion protein, followed by detection using commercially available GST antibodies. The nonradioactive approach is substantially more expensive (due to the purchase of antibody and detection reagents) than using radioactively labeled proteins. In addition, care must be taken to control for nonspecific interactions with GST alone and a signal resulting from antibody cross-reactivity. In some instances, proteins on the membrane are not able to interact after transfer. This may be due to improper folding, particularly in the case of proteins expressed from a phage expression library. This protocol describes a way to overcome this by washing the membrane in denaturation buffer, which is then serially diluted to permit slow renaturation of the proteins.

  9. Utilizing harmonization and common surveillance methods to consolidate 4 cohorts: the Western Alaska Tribal Collaborative for Health (WATCH study

    Directory of Open Access Journals (Sweden)

    Kathryn R. Koller

    2013-05-01

    Full Text Available Background. According to health status reports, chronic disease prevalence appears to be rising in western Alaska Native (AN people, and accurate population-based data are needed. Four cohort studies of western AN people were conducted in the Norton Sound and Yukon-Kuskokwim regions, but none have been large enough to allow reliable estimates of rates of chronic diseases and evaluate their risk factors. Objective. In this article, the methods used to combine 4 major cohort studies of rural western AN people are described and the benefits and challenges encountered in combining data and standardizing surveillance methods for these studies are discussed. Design. Tribal permission was obtained for each cohort study and the consolidated study. Data from baseline exams were directly combined or harmonized into new variables. Common surveillance methods were developed and implemented to identify incidence and risk factors for cardiovascular disease (CVD events and type 2 diabetes. Results. A cohort of 4,569 western AN participants (2,116 men and 2,453 women, aged 18–95 years, was established to study CVD and diabetes prevalence. Prospective surveillance data over an average 6.7-year follow-up can now be used to study CVD and diabetes incidence and associated risk factors in a subset of 2,754 western AN participants (1,218 men and 1,536 women who consented to initial surveillance. Conclusions. The combined cohort provides statistical power to examine incidence rates and risk factors for CVD and diabetes and allows for analyses by geographic region. The data can be used to develop intervention programmes in these populations and others.

  10. A Simple and Sensitive Plant-Based Western Corn Rootworm Bioassay Method for Resistance Determination and Event Selection.

    Science.gov (United States)

    Wen, Zhimou; Chen, Jeng Shong

    2018-05-26

    We report here a simple and sensitive plant-based western corn rootworm, Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae), bioassay method that allows for examination of multiple parameters for both plants and insects in a single experimental setup within a short duration. For plants, injury to roots can be visually examined, fresh root weight can be measured, and expression of trait protein in plant roots can be analyzed. For insects, in addition to survival, larval growth and development can be evaluated in several aspects including body weight gain, body length, and head capsule width. We demonstrated using the method that eCry3.1Ab-expressing 5307 corn was very effective against western corn rootworm by eliciting high mortality and significantly inhibiting larval growth and development. We also validated that the method allowed determination of resistance in an eCry3.1Ab-resistant western corn rootworm strain. While data presented in this paper demonstrate the usefulness of the method for selection of events of protein traits and for determination of resistance in laboratory populations, we envision that the method can be applied in much broader applications.

  11. Application of field methods to assess isometamidium resistance of trypanosomes in cattle in western Ethiopia

    DEFF Research Database (Denmark)

    Tewelde, N.; Abebe, G.; Eisler, M.

    2004-01-01

    This study assessed the degree of isometamidium resistance of trypanosomes infecting cattle in the upper Didessa valley of western Ethiopia. An initial prevalence study was conducted to identify sites with a high risk of trypanosmosis in cattle. The trypanosome prevalence varied widely, with two...

  12. Pattern Classification of Tropical Cyclone Tracks over the Western North Pacific using a Fuzzy Clustering Method

    Science.gov (United States)

    Kim, H.; Ho, C.; Kim, J.

    2008-12-01

    This study presents the pattern classification of tropical cyclone (TC) tracks over the western North Pacific (WNP) basin during the typhoon season (June through October) for 1965-2006 (total 42 years) using a fuzzy clustering method. After the fuzzy c-mean clustering algorithm to the TC trajectory interpolated into 20 segments of equivalent length, we divided the whole tracks into 7 patterns. The optimal number of the fuzzy cluster is determined by several validity measures. The classified TC track patterns represent quite different features in the recurving latitudes, genesis locations, and geographical pathways: TCs mainly forming in east-northern part of the WNP and striking Korean and Japan (C1); mainly forming in west-southern part of the WNP, traveling long pathway, and partly striking Japan (C2); mainly striking Taiwan and East China (C3); traveling near the east coast of Japan (C4); traveling the distant ocean east of Japan (C5); moving toward South China and Vietnam straightly (C6); and forming in the South China Sea (C7). Atmospheric environments related to each cluster show physically consistent with each TC track patterns. The straight track pattern is closely linked to a developed anticyclonic circulation to the north of the TC. It implies that this ridge acts as a steering flow forcing TCs to move to the northwest with a more west-oriented track. By contrast, recurving patterns occur commonly under the influence of the strong anomalous westerlies over the TC pathway but there definitely exist characteristic anomalous circulations over the mid- latitudes by pattern. Some clusters are closely related to the well-known large-scale phenomena. The C1 and C2 are highly related to the ENSO phase: The TCs in the C1 (C2) is more active during La Niña (El Niño). The TC activity in the C3 is associated with the WNP summer monsoon. The TCs in the C4 is more (less) vigorous during the easterly (westerly) phase of the stratospheric quasi-biennial oscillation

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

    International Nuclear Information System (INIS)

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

    1985-01-01

    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

  14. SPECIFIC CHARACTERISTICS OF IMPLEMENTING THE COMBINED METHOD OF TEACHING FOREIGN LANGUAGES IN WESTERN UKRAINE AND IN RUSSIA (1918–1939

    Directory of Open Access Journals (Sweden)

    Богдана Лабінська

    2014-04-01

    Full Text Available The paper investigates the specific characteristics of implementing the combined method in teaching foreign languages in Western Ukraine and in Russia (1918–1939. Preconditions of the origin of the combined method have been ascertained. Teaching language aspects and communicative activities has been analyzed on the basis of curricula, manuals and scientific articles of those times. Comparative studies of foreign language teaching methodology in Ukraine and in Russia have been carried out. Fundamentals of the combined method of teaching foreign languages have been substantiated and peculiarities of their realization in the region under investigation have been formulated.

  15. Northern blot analysis to investigate the abundance of microorganisms

    International Nuclear Information System (INIS)

    Krause, D.O.

    2005-01-01

    areas known as hyper-variable regions which have a high degree of sequence variation. As a result of this structure, it is possible to design signature oligonucleotide probes varying in length from about 15 to 30 nucleotides that are diagnostic of microorganisms at the kingdom, domain, genus and even species level. These signature sequences can be used in a variety of applications such as PCR analysis, construction of clone libraries or direct probing of bulk rRNA. In this chapter, I provide detailed protocols for the analysis of extracted rRNA and give detailed procedures that must be followed to do northern blot analysis of bulk RNA extracted from the rumen

  16. Combining X-ray based methods to study the protohistoric bronze technology in Western Iberia

    Energy Technology Data Exchange (ETDEWEB)

    Valério, P., E-mail: pvalerio@ctn.ist.utl.pt [Centro de Ciências e Tecnologias Nucleares (C2TN), Instituto Superior Técnico, Universidade de Lisboa, Campus Tecnológico e Nuclear, Estrada Nacional 10 (km 139,7), 2695-066 Bobadela LRS (Portugal); Silva, R.J.C., E-mail: rjcs@fct.unl.pt [CENIMAT/I3N, Departamento de Ciência dos Materiais, Faculdade de Ciências e Tecnologia, FCT, Universidade Nova de Lisboa, 2829-516 Monte de Caparica (Portugal); Soares, A.M.M., E-mail: amsoares@ctn.ist.utl.pt [Centro de Ciências e Tecnologias Nucleares (C2TN), Instituto Superior Técnico, Universidade de Lisboa, Campus Tecnológico e Nuclear, Estrada Nacional 10 (km 139,7), 2695-066 Bobadela LRS (Portugal); Araújo, M.F., E-mail: faraujo@ctn.ist.utl.pt [Centro de Ciências e Tecnologias Nucleares (C2TN), Instituto Superior Técnico, Universidade de Lisboa, Campus Tecnológico e Nuclear, Estrada Nacional 10 (km 139,7), 2695-066 Bobadela LRS (Portugal); Gonçalves, A.P., E-mail: apg@ctn.ist.utl.pt [Centro de Ciências e Tecnologias Nucleares (C2TN), Instituto Superior Técnico, Universidade de Lisboa, Campus Tecnológico e Nuclear, Estrada Nacional 10 (km 139,7), 2695-066 Bobadela LRS (Portugal); Soares, R.M., E-mail: ruigusmao@hotmail.com [UNIARQ, Centro de Arqueologia da Universidade de Lisboa, Faculdade de Letras, Alameda da Universidade, 1600-214 Lisboa (Portugal)

    2015-09-01

    The Phoenician arrival at Iberian coastal regions had an actual influence on indigenous technology. A collection of coeval metallurgical remains and artefacts was studied by EDXRF, micro-EDXRF, SEM–EDS and XRD, to identify certain features of the production and utilisation of metal in protohistoric Western Iberia. The composition of artefacts indicates a prevalence of Cu–Sn alloys with low content of impurities (Pb, As, Sb and Fe) during Late Bronze and Early Iron Ages, while the composition of slags points to a smaller loss of copper in Phoenician smelting operations. Moreover, the amount of iron impurities in metal proved to be a helpful discriminator between indigenous and Phoenician-based metallurgies, showing that later alloys have higher amounts of iron. Besides, the indigenous alloys have higher tin contents that can probably be explained by the easier access to metal sources of local communities.

  17. Combining X-ray based methods to study the protohistoric bronze technology in Western Iberia

    International Nuclear Information System (INIS)

    Valério, P.; Silva, R.J.C.; Soares, A.M.M.; Araújo, M.F.; Gonçalves, A.P.; Soares, R.M.

    2015-01-01

    The Phoenician arrival at Iberian coastal regions had an actual influence on indigenous technology. A collection of coeval metallurgical remains and artefacts was studied by EDXRF, micro-EDXRF, SEM–EDS and XRD, to identify certain features of the production and utilisation of metal in protohistoric Western Iberia. The composition of artefacts indicates a prevalence of Cu–Sn alloys with low content of impurities (Pb, As, Sb and Fe) during Late Bronze and Early Iron Ages, while the composition of slags points to a smaller loss of copper in Phoenician smelting operations. Moreover, the amount of iron impurities in metal proved to be a helpful discriminator between indigenous and Phoenician-based metallurgies, showing that later alloys have higher amounts of iron. Besides, the indigenous alloys have higher tin contents that can probably be explained by the easier access to metal sources of local communities

  18. Glycosaminoglycan blotting on nitrocellulose membranes treated with cetylpyridinium chloride after agarose-gel electrophoretic separation.

    Science.gov (United States)

    Maccari, Francesca; Volpi, Nicola

    2002-09-01

    We describe a method for blotting and immobilizing several nonsulfated and sulfated complex polysaccharides on membranes made hydrophilic and positively charged by a cationic detergent after their separation by conventional agarose gel electrophoresis. Nitrocellulose membranes were derivatized with the cationic detergent cetylpyridinium chloride (CPC) and mixtures of glycosaminoglycans (GAGs) were capillary-blotted after their separation in agarose gel electrophoresis in barium acetate/1,2-diaminopropane. Single purified species of variously sulfated polysaccharides were transferred onto the derivatized membranes after electrophoresis with an efficiency of 100% and stained with alcian blue (irreversible staining) and toluidine blue (reversible staining) permitting about 0.1 nug threshold of detection. Nonsulfated polyanions, hyaluronic acid, a fructose-containing polysaccharide with a chondroitin backbone purified from Escherichia coli U1-41, and its defructosylated product, were also electrophoretically separated and transferred onto membranes. The limit of detection for desulfated GAGs was about 0.1-0.5 nug after irreversible or reversible staining. GAG extracts from bovine, lung and aorta, and human aorta and urine were separated by agarose gel electrophoresis and blotted on CPC-treated nitrocellulose membranes. The polysaccharide composition of these extracts was determined. The membrane stained with toluidine blue (reversible staining) was destained and the same lanes used for immunological detection or other applications. Reversible staining was also applied to recover single species of polysaccharides after electrophoretic separation of mixtures of GAGs and their transfer onto membranes. Single bands were released from the membrane with an efficiency of 70-100% for further biochemical characterization.

  19. Crustal Models Assessment in Western Part of Romania Employing Active Seismic and Seismologic Methods

    Science.gov (United States)

    Bala, Andrei; Toma-Danila, Dragos; Tataru, Dragos; Grecu, Bogdan

    2017-12-01

    In the years 1999 - 2000 two regional seismic refraction lines were performed within a close cooperation with German partners from University of Karlsruhe. One of these lines is Vrancea 2001, with 420 km in length, almost half of them recorded in Transylvanian Basin. The structure of the crust along the seismic line revealed a very complicated crustal structure beginning with Eastern Carpathians and continuing in the Transylvanian Basin until Medias. As a result of the development of the National Seismic Network in the last ten years, more than 100 permanent broadband stations are now continuously operating in Romania. Complementary to this national dataset, maintained and developed in the National Institute for Earth Physics, new data emerged from the temporary seismologic networks established during the joint projects with European partners in the last decades. The data gathered so far is valuable both for seismology purposes and crustal structure studies, especially for the western part of the country, where this kind of data were sparse until now. Between 2009 and 2011, a new reference model for the Earth’s crust and mantle of the European Plate was defined through the NERIES project from existing data and models. The database gathered from different kind of measurements in Transylvanian Basin and eastern Pannonian Basin were included in this NERIES model and an improved and upgraded model of the Earth crust emerged for western part of Romania. Although the dataset has its origins in several periods over the last 50 years, the results are homogeneous and they improve and strengthen our image about the depth of the principal boundaries in the crust. In the last chapter two maps regarding these boundaries are constructed, one for mid-crustal boundary and one for Moho. They were build considering all the punctual information available from different sources in active seismic and seismology which are introduced in the general maps from the NERIES project for

  20. Unusual Rebuilding Method of Historic St Mary's Cathedral in the Capital of Western Australia

    Science.gov (United States)

    Wysokowski, Adam

    2016-06-01

    St Mary's Cathedral is the Archbishop church of the Archdiocese in Perth in Western Australia. The presented sacral building was built in neo-Gothic style during the years 1863-1865. Cathedral was officially dedicated and opened for the service on 29th January, 1865. In 1973 was proclaimed the Marian Sanctuary and now represents one of the largest religious facilities in Perth. In 2005, the city authorities, together with the Archdiocese took a collective decision on the necessity of a comprehensive renovation of this sacred object. The renovation was due to the technical condition and the lack of usability of the object. The author of the paper had the opportunity to experience these problems by visiting this place several times, first time in 1989 and next years. Thus, the renovation of the present Cathedral was in its assumption not only to perform maintenance of the building and its specific architectural elements but also to increase its functional features - usable for the faithful and tourists. Reconstruction of St Mary's Cathedral in Perth can be a good example increasing the wider functionality of such facilities while keeping their antique and historical qualities. In this paper the above-mentioned issues will be more widely developed by the author.

  1. Detection of alien genetic introgressions in bread wheat using dot-blot genomic hybridisation.

    Science.gov (United States)

    Rey, María-Dolores; Prieto, Pilar

    2017-01-01

    Simple, reliable methods for the identification of alien genetic introgressions are required in plant breeding programmes. The use of genomic dot-blot hybridisation allows the detection of small Hordeum chilense genomic introgressions in the descendants of genetic crosses between wheat and H. chilense addition or substitution lines in wheat when molecular markers are difficult to use. Based on genomic in situ hybridisation, DNA samples from wheat lines carrying putatively H. chilense introgressions were immobilised on a membrane, blocked with wheat genomic DNA and hybridised with biotin-labelled H. chilense genomic DNA as a probe. This dot-blot screening reduced the number of plants necessary to be analysed by molecular markers or in situ hybridisation, saving time and money. The technique was sensitive enough to detect a minimum of 5 ng of total genomic DNA immobilised on the membrane or about 1/420 dilution of H. chilense genomic DNA in the wheat background. The robustness of the technique was verified by in situ hybridisation. In addition, the detection of other wheat relative species such as Hordeum vulgare , Secale cereale and Agropyron cristatum in the wheat background was also reported .

  2. Influence of DNA treatments on Southern blot hybridization analysis ...

    African Journals Online (AJOL)

    STORAGESEVER

    2008-06-03

    Jun 3, 2008 ... DNA samples obtained by a non-phenol/chloroform isolation method, from three races of Fusarium oxysporum f. sp. lycopersici ... Key words: Fusarium oxysporum, DIG-IGS Probe, Southern hybridization. INTRODUCTION .... Detection of Fusarium spp in plants with monoclonal antibody. Ann. Phytopathol.

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

    Directory of Open Access Journals (Sweden)

    Bowlin Gary L

    2007-10-01

    Full Text Available Abstract Background Electrospinning is a non-mechanical processing strategy that can be used to process a variety of native and synthetic polymers into highly porous materials composed of nano-scale to micron-scale diameter fibers. By nature, electrospun materials exhibit an extensive surface area and highly interconnected pore spaces. In this study we adopted a biological engineering approach to ask how the specific unique advantages of the electrospinning process might be exploited to produce a new class of research/diagnostic tools. Methods The electrospinning properties of nitrocellulose, charged nylon and blends of these materials are characterized. Results Nitrocellulose electrospun from a starting concentration of Conclusion The flexibility afforded by electrospinning process makes it possible to tailor blotting membranes to specific applications. Electrospinning has a variety of potential applications in the clinical diagnostic field of use.

  4. Standardizing operational vector sampling techniques for measuring malaria transmission intensity: evaluation of six mosquito collection methods in western Kenya.

    Science.gov (United States)

    Wong, Jacklyn; Bayoh, Nabie; Olang, George; Killeen, Gerry F; Hamel, Mary J; Vulule, John M; Gimnig, John E

    2013-04-30

    Operational vector sampling methods lack standardization, making quantitative comparisons of malaria transmission across different settings difficult. Human landing catch (HLC) is considered the research gold standard for measuring human-mosquito contact, but is unsuitable for large-scale sampling. This study assessed mosquito catch rates of CDC light trap (CDC-LT), Ifakara tent trap (ITT), window exit trap (WET), pot resting trap (PRT), and box resting trap (BRT) relative to HLC in western Kenya to 1) identify appropriate methods for operational sampling in this region, and 2) contribute to a larger, overarching project comparing standardized evaluations of vector trapping methods across multiple countries. Mosquitoes were collected from June to July 2009 in four districts: Rarieda, Kisumu West, Nyando, and Rachuonyo. In each district, all trapping methods were rotated 10 times through three houses in a 3 × 3 Latin Square design. Anophelines were identified by morphology and females classified as fed or non-fed. Anopheles gambiae s.l. were further identified as Anopheles gambiae s.s. or Anopheles arabiensis by PCR. Relative catch rates were estimated by negative binomial regression. When data were pooled across all four districts, catch rates (relative to HLC indoor) for An. gambiae s.l (95.6% An. arabiensis, 4.4% An. gambiae s.s) were high for HLC outdoor (RR = 1.01), CDC-LT (RR = 1.18), and ITT (RR = 1.39); moderate for WET (RR = 0.52) and PRT outdoor (RR = 0.32); and low for all remaining types of resting traps (PRT indoor, BRT indoor, and BRT outdoor; RR < 0.08 for all). For Anopheles funestus, relative catch rates were high for ITT (RR = 1.21); moderate for HLC outdoor (RR = 0.47), CDC-LT (RR = 0.69), and WET (RR = 0.49); and low for all resting traps (RR < 0.02 for all). At finer geographic scales, however, efficacy of each trap type varied from district to district. ITT, CDC-LT, and WET appear to be effective methods for large-scale vector sampling in

  5. Heard but not seen: Comparing bat assemblages and study methods in a mosaic landscape in the Western Ghats of India.

    Science.gov (United States)

    Wordley, Claire F R; Sankaran, Mahesh; Mudappa, Divya; Altringham, John D

    2018-04-01

    We used capture (mist-netting) and acoustic methods to compare the species richness, abundance, and composition of a bat assemblage in different habitats in the Western Ghats of India. In the tropics, catching bats has been more commonly used as a survey method than acoustic recordings. In our study, acoustic methods based on recording echolocation calls detected greater bat activity and more species than mist-netting. However, some species were detected more frequently or exclusively by capture. Ideally, the two methods should be used together to compensate for the biases in each. Using combined capture and acoustic data, we found that protected forests, forest fragments, and shade coffee plantations hosted similar and diverse species assemblages, although some species were recorded more frequently in protected forests. Tea plantations contained very few species from the overall bat assemblage. In riparian habitats, a strip of forested habitat on the river bank improved the habitat for bats compared to rivers with tea planted up to each bank. Our results show that shade coffee plantations are better bat habitat than tea plantations in biodiversity hotspots. However, if tea is to be the dominant land use, forest fragments and riparian corridors can improve the landscape considerably for bats. We encourage coffee growers to retain traditional plantations with mature native trees, rather than reverting to sun grown coffee or coffee shaded by a few species of timber trees.

  6. Developing and validating a perinatal depression screening tool in Kenya blending Western criteria with local idioms: A mixed methods study.

    Science.gov (United States)

    Green, Eric P; Tuli, Hawa; Kwobah, Edith; Menya, D; Chesire, Irene; Schmidt, Christina

    2018-03-01

    Routine screening for perinatal depression is not common in most primary health care settings. The U.S. Preventive Services Task Force only recently updated their recommendation on depression screening to specifically recommend screening during the pre- and postpartum periods. While practitioners in high-income countries can respond to this new recommendation by implementing one of several existing depression screening tools developed in Western contexts, such as the Edinburgh Postnatal Depression Scale (EPDS) or the Patient Health Questionnaire-9 (PHQ-9), these tools lack strong evidence of cross-cultural equivalence, validity for case finding, and precision in measuring response to treatment in developing countries. Thus, there is a critical need to develop and validate new screening tools for perinatal depression that can be used by lay health workers, primary health care personnel, and patients. Working in rural Kenya, we used free listing, card sorting, and item analysis methods to develop a locally-relevant screening tool that blended Western psychiatric concepts with local idioms of distress. We conducted a validation study with a random sample of 193 pregnant women and new mothers to test the diagnostic accuracy of this scale along with the EPDS and PHQ-9. The sensitivity/specificity of the EPDS and PHQ-9 was estimated to be 0.70/0.72 and 0.70/0.73, respectively. This compared to sensitivity/specificity of 0.90/0.90 for a new 9-item locally-developed tool called the Perinatal Depression Screening (PDEPS). Across these three tools, internal consistency reliability ranged from 0.77 to 0.81 and test-retest reliability ranged from 0.57 to 0.67. The prevalence of depression ranges from 5.2% to 6.2% depending on the clinical reference standard. The EPDS and PHQ-9 are valid and reliable screening tools for perinatal depression in rural Western Kenya, the PDEPS may be a more useful alternative. At less than 10%, the prevalence of depression in this region appears

  7. Investigation of the shelf break and continental slope in the Western part of the Black Sea using acoustic methods

    Science.gov (United States)

    Dutu, F.; Ion, G.; Jugaru Tiron, L.

    2009-04-01

    The Black Sea is a large marginal sea surrounded by a system of Alpine orogenic chains, including the Balkanides-Pontides, Caucasus, Crimea and North Dobrogea located to the south, northeast, north and northwest, respectively (Dinu et al., 2005). The north-western part of the Black Sea is the main depocentre for sediment supply from Central Europe via the Danube River, but also from Eastern Europe through the Ukrainian rivers Dniepr, Dniestr and Southern Bug (Popescu et al., 2004). The shelfbreak is located at water depths of 120-140 m southward of the Danube Canyon, and up to 170 m northward of the canyon possibly due to recent faulting which is very common in this area. The continental slope is dissected by numerous canyons, each of which is fed by several tributaries. The Danube Canyon (also known as Viteaz Canyon) is a large shelf-indenting canyon located in the north-western Black Sea and connected to the youngest channel-levee system of the Danube Fan (Popescu et al., 2004). The acoustic methods are a useful way for investigate the shelf break and the continental slope giving us information about landslides on the continental slope, the topography of the investigated area, the sedimentary zones affected by instability and to quantify the geometry of the underwater landslides. The measurements made on the continental slope from north-western part of the Black Sea gave us the possibility to make a digital terrain model. After processing the data the model offer information about the main access ways of the sediments through gravitational slide on the submarines canyons, with forming of turbidity currents, debris flows and also other transport/transformation phenomena of the sediments on the continental slope like submarine landslides and submarine collapse. References Dinu, C., Wong, H.K., Tambrea, D., Matenco, L., 2005. Stratigraphic and structural characteristics of the Romanian Black Sea shelf. Tectonophysics 410, 417-435. Popescu, I., Lericolais, G., Panin

  8. Depression literacy and health-seeking attitudes in the Western Pacific region: a mixed-methods study.

    Science.gov (United States)

    Ho, Grace W K; Bressington, D; Leung, S F; Lam, K K C; Leung, A Y M; Molassiotis, A; Ligot, J; Ranoco, C; Sophal, C; Valimaki, M

    2018-06-02

    Depression literacy refers to the ability to recognize depression and make informed decisions about its treatment. To date, relatively little research has been done to examine depression literacy in the Western Pacific region. Given the pervasiveness of depression and the need to enhance mental health care in this region, it is important to gain a better understanding of depression literacy and health-seeking behaviors in this part of the world. This mixed-methods study utilized a convergent parallel design to examine depression literacy and the associated health-seeking attitudes among urban adults from three countries-Cambodia, Philippines, and Fiji. A total of 455 adults completed a quantitative survey on depression knowledge, attitudes, and professional help seeking. Separately, 56 interviewees from 6 focus groups provided qualitative data on their impression and knowledge of depression and mental illness within the context of their local communities. Overall, results showed that depression knowledge was comparatively lower in this region. Controlling for differences across countries, higher knowledge was significantly associated with more positive attitudes towards mental illness (B = - 0.28, p = 0.025) and professional help seeking (B = 0.20, p < 0.001). Financial stability, such as employment, was also a salient factor for help seeking. This study was the first to provide a baseline understanding on depression literacy and highlights the need to increase public knowledge on depression in the Western Pacific. Culturally congruent recommendations on enhancing depression literacy in this region, such as anti-stigma campaigns, use of financial incentives, and family-based approach in health education, are discussed.

  9. Use of Low-Cost Methods of Soil Erosion Control In Kisii District, South Western kenya

    International Nuclear Information System (INIS)

    Nzabi, A.W; Makini, F; Onyango, M; Mureithi, J.G

    1999-01-01

    Kisii District has a topography of undulating hills and is prone to severe soil erosion. The average rainfall is 1900 mm and occurs in biomodal pattern. During a participatory appraisal survey in 1995, farmers indicated that soil erosion in the area had contributed to decline in soil fertility resulting in low crop yields. To address this problem, an on-farm trial was conducted in 1996 at Nyamonyo village to test the effectiveness of four low cost methods of controlling soil erosion. These included maize stover trash line, sweet potatoes,Penicum maximum var. Makarikari grass strip and vetiveria zizanioides (Vertiver) grass strip. A treatment without soil erosion control measure was included. The trial was planted in three farms which acted as replicates. The treatments were planted in runoff plots measuring 4 x 2 m in which had a maize crop were laid down in a randomized complete block design. Surface runoff and eroded soils were collected in 50-l buckets. The experimental site had a slope ranging from 16 to 35%. Preliminary results indicated that maize stover trash line and sweet potato strips were more effective in controlling soil erosion than the grass strips. As the season progressed the grass strips became increasingly more effective in erosion control. The trail is still continuing but results indicate that for short term soil erosion control, maize stover trash lines and sweet potatoes are more effective while Makarikari and Vertiver grass strips are promising as long term soil erosion control measure

  10. Immunological diagnosis of human hydatid cyst using Western immunoblotting technique

    Directory of Open Access Journals (Sweden)

    Mahboubeh Hadipour

    2016-01-01

    Full Text Available Background: Echinococcosis is a parasitic disease with worldwide distribution which is caused by the tapeworms Echinococcus granulosus. Diagnosis of the disease relies on imaging techniques, but the techniques are not able to differentiate the cyst from benign or malignant tumors; hence, appropriate serologic methods are required for the differential diagnosis of the infection. Materials and Methods: In this investigation, different sheep hydatid cyst antigens probed with thirty sera of patients with hydatid cyst and also thirty human normal sera using Western immunoblotting technique. Considering results of surgery as gold standard, sensitivity and specificity of Western blotting was estimated. Results: Sera of 29, 26, and 16 patients with hydatid cyst reacted with specific bands of hydatid cyst fluid (HCF, protoscolex crude antigen, and cyst wall crude antigen, respectively. However, none of the normal human sera reacted with those specific bands. Conclusion: A 20 kDa band of sheep HCF is an appropriate antigen for serodiagnosis of hydatid cyst infection.

  11. Aerosol optical depth in a western Mediterranean site: An assessment of different methods

    Science.gov (United States)

    Sanchez-Romero, A.; González, J. A.; Calbó, J.; Sanchez-Lorenzo, A.; Michalsky, J.

    2016-06-01

    Column aerosol optical properties were derived from multifilter rotating shadowing radiometer (MFRSR) observations carried out at Girona (northeast Spain) from June 2012 to June 2014. We used a technique that allows estimating simultaneously aerosol optical depth (AOD) and Ångström exponent (AE) at high time-resolution. For the period studied, mean AOD at 500 nm was 0.14, with a noticeable seasonal pattern, i.e. maximum in summer and minimum in winter. Mean AE from 500 to 870 nm was 1.2 with a strong day-to-day variation and slightly higher values in summer. So, the summer increase in AOD seems to be linked with an enhancement in the number of fine particles. A radiative closure experiment, using the SMARTS2 model, was performed to confirm that the MFRSR-retrieved aerosol optical properties appropriately represent the continuously varying atmospheric conditions in Girona. Thus, the calculated broadband values of the direct flux show a mean absolute difference of less than 5.9 W m- 2 (0.77%) and R = 0.99 when compared to the observed fluxes. The sensitivity of the achieved closure to uncertainties in AOD and AE was also examined. We use this MFRSR-based dataset as a reference for other ground-based and satellite measurements that might be used to assess the aerosol properties at this site. First, we used observations obtained from a 100 km away AERONET station; despite a general similar behavior when compared with the in-situ MFRSR observations, certain discrepancies for AOD estimates in the different channels (R < 0.84 and slope < 1) appear. Second, AOD products from MISR and MODIS satellite observations were compared with our ground-based retrievals. Reasonable agreements are found for the MISR product (R = 0.92), with somewhat poorer agreement for the MODIS product (R = 0.70). Finally, we apply all these methods to study in detail the aerosol properties during two singular aerosol events related to a forest fire and a desert dust intrusion.

  12. Modification of T-cell antigenic properties of tetanus toxoid by SDS-PAGE separation. Implications for T-cell blotting

    DEFF Research Database (Denmark)

    Christensen, C B; Theander, T G

    1997-01-01

    Using Tetanus Toxoid (TT) as a model antigen the T-cell Blotting method was evaluated. Peripheral blood mononuclear cell (PBMC) cultures were stimulated by blotted nitrocellulose-bound TT or soluble TT. SDS-Poly-Acrylamide-Gel-Electrophoresis separated TT only induced proliferation in 20% of the ......Using Tetanus Toxoid (TT) as a model antigen the T-cell Blotting method was evaluated. Peripheral blood mononuclear cell (PBMC) cultures were stimulated by blotted nitrocellulose-bound TT or soluble TT. SDS-Poly-Acrylamide-Gel-Electrophoresis separated TT only induced proliferation in 20......% of the PBMC cultures whereas proliferation was induced in 79% of the same cultures offered similar treated TT (except for the PAGE separation). When T-cell blotting was performed with TT separated in a SDS-agarose matrix, proliferation was induced in 80% of donors responding to soluble TT. The results show...... that SDS-PAGE alters the ability of TT to induce T-cell proliferation, possibly due to unpolymerized acrylamide binding to proteins during SDS-PAGE. The use of SDS-PAGE T-cell blotting in the screening for T-cell antigens must therefore be reconsidered. We suggest the use of SDS-Agarose Gel Electrophoresis...

  13. Use of nitrocellulose blotting for the study of hepatitis B surface antigen electrophoresed in agarose gels

    Energy Technology Data Exchange (ETDEWEB)

    McMichael, J C; Greisiger, L M; Millman, I [Institute for Cancer Research, Philadelphia, PA (USA). Fox Chase Cancer Center

    1981-08-28

    Nitrocellulose-protein blotting of serum electrophoresed in agarose gels has been adapted for the study of hepatitis B surface antigen (HBsAg). /sup 125/I-labeled anti-HBs was used as the antigen probe, and the electrophoretic migration was monitored by autoradiography. The method required 3 ..mu..l or less of serum and could detect as little as 1 pg of purified HBsAg. Typically, the authors observed two bands of HBsAg; a moving band which migrated about one-third the distance moved by human serum albumin and a non-migratory band which remained at the loading site. Some examples of the use of the method include: (1) empirical methods for correlating HBsAg concentration in serum to film darkness; (2) observations of mobility changes in serial sera from dialysis patients with chronic HBsAg antigenemia; and (3) detection of related antigens such as antigen from the PLC/PRF/5 hepatoma tissue culture line and the cross-reacting woodchuck hepatitis virus surface antigen (WHsAg).

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

    Directory of Open Access Journals (Sweden)

    Eikan Mishima

    Full Text Available 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.

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

    Science.gov (United States)

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

    2015-01-01

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

  16. The efficacy of permethrin-treated bed nets on child mortality and morbidity in western Kenya II. Study design and methods

    NARCIS (Netherlands)

    Phillips-Howard, Penelope A.; ter Kuile, Feiko O.; Nahlen, Bernard L.; Alaii, Jane A.; Gimnig, John E.; Kolczak, Margarette S.; Terlouw, Dianne J.; Kariuki, Simon K.; Shi, Ya Ping; Kachur, S. Patrick; Hightower, Allen W.; Vulule, John M.; Hawley, William A.

    2003-01-01

    This paper describes the study design and methods used in a large community-based, group-randomized, controlled trial of permethrin-treated bed nets (ITNs) in an area with intense, perennial malaria transmission in western Kenya conducted between 1996 and 1999. A multi-disciplinary framework was

  17. Methods to uncover an antibody epitope in the KPI domain of Alzheimer's amyloid precursor protein for immunohistochemistry in human brain.

    Science.gov (United States)

    Campbell, E; Pearson, R C; Parkinson, D

    1999-11-15

    A novel polyclonal antibody (Ab993), specific for a KPI domain epitope of APP, was characterised for use in immunoprecipitation, Western blotting and immunohistochemistry. Conditioned medium from NTera2/D1 cells was used for immunoprecipitation and Western blots. Paraffin-embedded human brain sections were used for immunohistochemistry. The antibody recognised KPI-containing APP on Western blots after standard solubilisation but immunoprecipitation of soluble APP required reduction with 2-mercaptoethanol followed by alkylation of reduced sulphydryl bonds with sodium iodoacetate. Immunohistochemical staining of human brain sections was significantly enhanced by this pre-treatment. Microwaving of sections also increased immunolabelling, by a mechanism that was additive to reduction and alkylation. Incubation in 80% formic acid did not confer any enhancement of immunoreactivity. Ab993, applied with the methods reported here, is expected to be valuable in investigations of the pathogenesis of Alzheimer's disease to determine the source of the beta-amyloid peptide.

  18. Western Sufism

    DEFF Research Database (Denmark)

    Sedgwick, Mark

    Western Sufism is sometimes dismissed as a relatively recent "new age" phenomenon, but in this book, Mark Sedgwick argues that it actually has very deep roots, both in the Muslim world and in the West. In fact, although the first significant Western Sufi organization was not established until 1915......, the first Western discussion of Sufism was printed in 1480, and Western interest in some of the ideas that are central to Sufi thought goes back to the thirteenth century. Sedgwick starts with the earliest origins of Western Sufism in late antique Neoplatonism and early Arab philosophy, and traces later......, the year in which the first Western Sufi order based not on the heritage of the European Middle Ages, Renaissance and Enlightenment, but rather on purely Islamic models, was founded. Later developments in this and other orders are also covered. Western Sufism shows the influence of these origins...

  19. Management practices associated with pain in cattle on western Canadian cow-calf operations: A mixed methods study.

    Science.gov (United States)

    Moggy, M A; Pajor, E A; Thurston, W E; Parker, S; Greter, A M; Schwartzkopf-Genswein, K S; Campbell, J R; Windeyer, M C

    2017-02-01

    The implementation of on-farm pain mitigation strategies is dependent on feasibility and importance to producers. Currently, there is a lack of information regarding adoption of management practices associated with pain in cattle within the Canadian beef industry. The objective of this mixed methods study was to describe pain-associated practices implemented on farm and producer perceptions toward pain mitigation strategies. A questionnaire about calving management and calf processing was delivered to 109 cow-calf producers in western Canada. In addition, 15 respondents were purposively selected based on questionnaire responses to participate in individual semistructured, on-farm interviews. The prevalence of pain mitigation strategies used for dystocia and cesarean section by respondents were 46 and 100%, respectively. The majority of operations reported castrating and dehorning calves before 3 mo of age (95 and 89%, respectively). The majority of operations did not use pain mitigation strategies for castration and dehorning (90 and 85%, respectively). Branding was practiced by 57% of respondents, 4% of which used pain mitigation. Thematic content analysis revealed that producers' perception of pain were influenced by what they referred to as "common sense," relatability to cattle, visual evidence of pain, and age of the animal. Factors that influenced participant rationale for the implementation of pain mitigation practices included access to information and resources, age of the animal, benefit to the operation, cost and logistics, market demands, and personal conscience. Overall, management practices were generally in compliance with published Canadian guidelines. Results of this study may provide direction for future policy making, research, and extension efforts to encourage the adoption of pain mitigation strategies.

  20. The varieties of contemplative experience: A mixed-methods study of meditation-related challenges in Western Buddhists.

    Directory of Open Access Journals (Sweden)

    Jared R Lindahl

    Full Text Available Buddhist-derived meditation practices are currently being employed as a popular form of health promotion. While meditation programs draw inspiration from Buddhist textual sources for the benefits of meditation, these sources also acknowledge a wide range of other effects beyond health-related outcomes. The Varieties of Contemplative Experience study investigates meditation-related experiences that are typically underreported, particularly experiences that are described as challenging, difficult, distressing, functionally impairing, and/or requiring additional support. A mixed-methods approach featured qualitative interviews with Western Buddhist meditation practitioners and experts in Theravāda, Zen, and Tibetan traditions. Interview questions probed meditation experiences and influencing factors, including interpretations and management strategies. A follow-up survey provided quantitative assessments of causality, impairment and other demographic and practice-related variables. The content-driven thematic analysis of interviews yielded a taxonomy of 59 meditation-related experiences across 7 domains: cognitive, perceptual, affective, somatic, conative, sense of self, and social. Even in cases where the phenomenology was similar across participants, interpretations of and responses to the experiences differed considerably. The associated valence ranged from very positive to very negative, and the associated level of distress and functional impairment ranged from minimal and transient to severe and enduring. In order to determine what factors may influence the valence, impact, and response to any given experience, the study also identified 26 categories of influencing factors across 4 domains: practitioner-level factors, practice-level factors, relationships, and health behaviors. By identifying a broader range of experiences associated with meditation, along with the factors that contribute to the presence and management of experiences reported

  1. Mammalian α-polymerase: cloning of partial complementary DNA and immunobinding of catalytic subunit in crude homogenate protein blots

    International Nuclear Information System (INIS)

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

    1987-01-01

    A new polyclonal antibody against the α-polymerase catalytic polypeptide was prepared by using homogeneous HeLa cellα-polymerase. The antibody neutralized α-polymerase activity and was strong and specific for the α-polymerase catalytic polypeptide (M/sub r/ 183,000) in Western blot analysis of crude extracts of HeLa cells. The antibody was used to screen a cDNA library of newborn rat brain poly(A+) RNA in λgt11. A positive phage was identified and plaque purified. This phage, designated λpolα1.2, also was found to be positive with an antibody against Drosophila α-polymerase. The insert in λpolα1.2 (1183 base pairs) contained a poly(A) sequence at the 3' terminus and a short in-phase open reading frame at the 5' terminus. A synthetic oligopeptide (eight amino acids) corresponding to the open reading frame was used to raise antiserum in rabbits. Antibody affinity purified from this serum was found to be immunoreactive against purified α-polymerase by enzyme-linked immunosorbent assay and was capable of immunoprecipitating α-polymerase. This indicated the λpolα1.2 insert encoded an α-polymerase epitope and suggested that the cDNA corresponded to an α-polymerase mRNA. This was confirmed in hybrid selection experiments using pUC9 containing the cDNA insert and poly(A+) RNA from newborn rat brain; the insert hybridized to mRNA capable of encoding α-polymerase catalytic polypeptides. Northern blot analysis of rat brain poly(A+) RNA revealed that this mRNA is ∼5.4 kilobases

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

    International Nuclear Information System (INIS)

    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 32 P dCTP and 32 P dATP to a specific activity greater then 1.0 x 10 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. Determinants of eating at local and western fast-food venues in an urban Asian population: a mixed methods approach.

    Science.gov (United States)

    Naidoo, Nasheen; van Dam, Rob M; Ng, Sheryl; Tan, Chuen Seng; Chen, Shiqi; Lim, Jia Yi; Chan, Mei Fen; Chew, Ling; Rebello, Salome A

    2017-05-25

    Like several Southeast Asian countries, Singapore has a complex eating-out environment and a rising eating-out prevalence. However the determinants and drivers of eating-out in urban Asian environments are poorly understood. We examined the socio-demographic characteristics of persons who frequently ate away from home in local eateries called hawker centres and Western fast-food restaurants, using data from 1647 Singaporean adults participating in the National Nutrition Survey (NNS) 2010. We also assessed the underlying drivers of eating out and evaluated if these were different for eating at local eateries compared to Western fast-food restaurants using 18 focus group discussions of women (130 women). Participants reported a high eating-out frequency with 77.3% usually eating either breakfast, lunch or dinner at eateries. Main venues for eating-out included hawker centres (61.1% usually ate at least 1 of 3 daily meals at this venue) and school/workplace canteens (20.4%). A minority of participants (1.9%) reported usually eating at Western fast-food restaurants. Younger participants and those of Chinese and Malay ethnicity compared to Indians were more likely to eat at Western fast-food restaurants. Chinese and employed persons were more likely to eat at hawker centres. The ready availability of a large variety of affordable and appealing foods appeared to be a primary driver of eating out, particularly at hawker centres. Our findings highlight the growing importance of eating-out in an urban Asian population where local eating venues play a more dominant role compared with Western fast-food chains. Interventions focusing on improving the food quality at venues for eating out are important to improve the diet of urban Asian populations.

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

    Science.gov (United States)

    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.

  5. [Analytic study of dot blotting for the detection of anti-Jo-1, anti-M2, anti-ribosomes and anti-LKM].

    Science.gov (United States)

    Huguet, S; Sghiri, R; Ballot, E; Johanet, C

    2004-01-01

    The Cyto-Dot 4 HM043 kit commercialised by BMD, has replaced the Cyto-Dot HM010 kit that allowed three auto-antibodies detection (anti-Jo-1, anti-M2 and anti-ribosomal protein). Detection of anti-LKM1 auto-antibody was added. These four auto-antibodies have in common only the intracytoplasmic localisation of their respective antigen. The aim of our study was to evaluate this new kit using 104 sera and to compare our results with reference techniques (indirect immunofluorescence IF for anti-M2, anti-ribosomal protein and anti-LKM1, double immunodiffusion ID for anti-Jo-1 and anti-LKM1, western blotting WB for anti-M2) and with Cyto-Dot HM010. The one hundred and four sera were divided into five groups: Group I (n = 12) with anti-Jo-1 detected by ID; Group II (n = 28) with 26 anti-M2 positive by IF and WB, 2 anti-M2 positive only by WB; Group III (n = 10) with anti-ribosomal protein detected by IF 5 of which precipitated by ID; Group IV (n = 32) with anti-LKM1 by IF and ID divided into 18 AIH2 and 14 HCV; Group V (n = 22) consisting of 14 healthy individuals and 8 patients with hypergammaglobulinemia. Results of this study are similar to those of Cyto-Dot HM010 for the three auto-antibodies already in use. Cyto-Dot 4 is a very good anti-LKM1 confirmation method as it is ID. Copyright John Libbey Eurotext 2003.

  6. Computer Simulation Western

    International Nuclear Information System (INIS)

    Rasmussen, H.

    1992-01-01

    Computer Simulation Western is a unit within the Department of Applied Mathematics at the University of Western Ontario. Its purpose is the development of computational and mathematical methods for practical problems in industry and engineering and the application and marketing of such methods. We describe the unit and our efforts at obtaining research and development grants. Some representative projects will be presented and future plans discussed. (author)

  7. Northern and Southern blot analysis of human RNA and DNA in autopsy material

    DEFF Research Database (Denmark)

    Larsen, S; Rygaard, K; Asnaes, S

    1992-01-01

    was obtained less than two days postmortem. Histological examination showing slight or no autolysis and the presence of ribosomal bands after gel electrophoresis were both indicative parameters of RNA preservation. DNA was appropriate for Southern blotting when the tissue was obtained less than three to five...

  8. Testing for myositis specific autoantibodies: Comparison between line blot and immunoprecipitation assays in 57 myositis sera.

    Science.gov (United States)

    Cavazzana, Ilaria; Fredi, Micaela; Ceribelli, Angela; Mordenti, Cristina; Ferrari, Fabio; Carabellese, Nice; Tincani, Angela; Satoh, Minoru; Franceschini, Franco

    2016-06-01

    To analyze the performance of a line blot assay for the identification of autoantibodies in sera of patients affected by myositis, compared with immunoprecipitation (IP) as gold standard. 66 sera of patients with myositis (23 polymyositis, 8 anti-synthetase syndromes, 29 dermatomyositis and 6 overlap syndromes) were tested by commercial LB (Euroimmun, Lubeck, Germany); 57 sera were analyzed also by IP of K562 cell extract radiolabeled with (35)S-methionine. Inter-rater agreement was calculated with Cohen's k coefficient. Myositis-specific antibodies (MSA) were detected in 36/57 sera (63%) by IP and in 39/66 sera (59%) by LB. The most frequent MSA found by LB were anti-Jo1 and anti-Mi2 found in 15% (10/66) of sera, followed by anti-NXP2 and anti-SRP detected in 106% (7/66) of sera. Anti-TIF1gamma and anti-MDA5 were found in 6 (9%) and 5 sera (7.6%), respectively. A good agreement between methods was found only for anti-TIF1γ, anti-MDA5 and anti-NXP-2 antibodies, while a moderate agreement was estimated for anti-Mi2 and anti-EJ. By contrast, a high discordance rate for the detection of anti-Jo1 antibodies was evident (k: 0.3). Multiple positivity for MSA were found in 11/66 (17%) by LB and 0/57 by IP (p: 0001). Comparing the clinical features of these 11 sera, we found total discrepancies between assays in 3 sera (27.3%), a relative discrepancy due to the occurrence of one discordant autoantibody (not confirmed by IP) in 5 cases (45.5%) and a total discrepancy between LB and IP results, but with a relative concordance with clinical features were found in other 3 sera (27.3%). The semiquantitative results do not support the interpretation of the data. The use of LB assay allowed the detection of new MSA, such as anti-MDA5, anti-MJ and anti-TIF1gamma antibodies, previously not found with routine methods. However, the high prevalence of multiple positivities and the high discondant rate of anti-Jo1 antibodies could create some misinterpretation of the results from the

  9. Contribution of outstanding teachers from Western Ukraine of late XIX – early XX century to development of theory and methods of teaching history

    Directory of Open Access Journals (Sweden)

    Zinoviia Nahachevska

    2017-02-01

    Full Text Available The article provides analysis of theses from works of prominent representatives of educational thought in Western Ukraine of late XIX – early XX century, which are related to solving problems of teaching history in elementary and secondary schools in the region. The emphasis is placed on the actualization of content and methods of teaching by V. VilshanetskaZhukovetska, A. Kopystianskyi, M. Korduba, and K. Malytska.Key words: elementary and secondary schools (gymnasia, curricula, content, principles and methods of teaching history, school textbooks, anthropologization and humanization of history, study of local lore.

  10. Westerns fra hele verden

    DEFF Research Database (Denmark)

    Bjerre, Thomas Ærvold

    2014-01-01

    Om den amerikanske western, spaghettiwesterns, kommunistiske westerns og danske westerns - i forbindelse med Kristian Levrings The Salvation (2014).......Om den amerikanske western, spaghettiwesterns, kommunistiske westerns og danske westerns - i forbindelse med Kristian Levrings The Salvation (2014)....

  11. Western lowland gorilla diet and resource availability: new evidence, cross-site comparisons, and reflections on indirect sampling methods.

    Science.gov (United States)

    Doran, Diane M; McNeilage, Alastair; Greer, David; Bocian, Carolyn; Mehlman, Patrick; Shah, Natasha

    2002-11-01

    We describe the resource availability and diet of western lowland gorillas (Gorilla gorilla gorilla) from a new study site in the Central African Republic and Republic of Congo based on 3 years of study. The results, based on 715 fecal samples and 617 days of feeding trails, were similar to those reported from three other sites, in spite of differences in herb and fruit availability. Staple foods (consumed year-round) included high-quality herbs (Haumania), swamp herbs (when present), and a minimal diversity of fruit. A variety of fruits (average of 3.5 species per day and 10 per month) were selectively consumed; gorillas ignored some common fruits and incorporated rare fruits to a degree higher than predicted based on availability. During periods of fruit abundance, fruit constituted most of the diet. When succulent fruits were unavailable, gorillas used low-quality herbs (i.e., low-protein), bark, and more fibrous fruits as fallback foods. Fibrous fruit species, such as Duboscia macrocarpa and Klainedoxa gabonensis, were particularly important to gorillas at Mondika and other sites as fallbacks. The densities of these two species are similar across sites for which data are available, in spite of major differences in forest structure, suggesting they may be key species in determining gorilla density. No sex difference in diet was detected. Such little variation in western lowland gorilla diet across sites and between sexes was unexpected and may partly reflect limitations of indirect sampling. Copyright 2002 Wiley-Liss, Inc.

  12. Effects of Tithonia diversifolia, farmyard manure and urea, and phosphate fertiliser application methods on maize yields in western Kenya

    Directory of Open Access Journals (Sweden)

    Peter Asbon Opala

    2015-05-01

    Full Text Available Maize production in western Kenya is often limited by deficiencies of nitrogen and phosphorus. We assessed the effectiveness of Tithonia diversifolia green manure (tithonia, farmyard manure (FYM and urea as sources of nitrogen (N for maize when inorganic phosphorus (P fertiliser was either broadcast (BR or spot-placed in the planting hole (SP for two consecutive seasons; October to December of 1998 and April to August of 1999 at two sites; Nyabeda and Khwisero in western Kenya. A randomised complete block design with four replications was used. Maize yields were higher at Nyabeda and responded to P application better than at Khwisero. At the same N rate, tithonia and FYM were as effective as urea in increasing maize yields at both sites. There were no significant differences in maize yields when phosphate fertiliser was either BR or SP regardless of the N source used in the first season. However, in the second season, the residual yields for the BR treatments were consistently higher than those of the SP. Our results suggest that tithonia and FYM can substitute for urea as N sources and that fertiliser P should be broadcast and incorporated together with the organic materials at the time of planting to save on labour costs.

  13. Imaging and high-sensitivity quantification of chemiluminescent labeled DNA-blots

    International Nuclear Information System (INIS)

    Dorner, G.

    1997-01-01

    The present thesis has for objective the development of both, methods of DNA labeling by chemiluminescence (via the catalytic activity of the enzyme alkaline phosphatase - AP) and an appropriate imaging system. Offering a competitive alternative to the detection of classical radio-labels in molecular-biological experiments of the blotting type, this technique should permit the realization of quantitative studies of gene expression at ultra-high sensitivity necessary in particular for differential-screening experiments. To reach our aim. we separated the project into three different parts. In a first step an imager based on a liquid-nitrogen-cooled CCD coupled to a standard optics (50 mm/fl.2) has been installed and characterized. This system offers a sensitive area of up to 625 cm 2 , a spatial resolution of 0.3-1 mm (depending on the field of view) and a sensitivity sufficient to detect 10 fg/mm 2 labeled DNA. In a second part, the chemiluminescent light-generation process in solution has been investigated to optimize the parameters temperature. pH and concentration of the substrate as well as the enzyme. The substrate offering the highest light yield (CDP-Star in addition with the enhancer EMERALD II) allows quantification of AP down to 10 -15 M within a dynamic range of 10 4 in solution. Finally. preparation, immobilization and detection of AP-labeled DNA probes (via a biotin-streptavidin-biotin-AP bridge) on nylon membranes has been optimized. A linear relation between the light intensities and the amount of DNA was observed in a range of 10 fg/mm 2 - 100 pg/mm 2 . Hybridization of the probes to bacterial cloned target-DNA has been addressed after examination of the best hybridization conditions. Our protocol includes the treatment of a proteinase, which resulted in a significantly lower background on the filter. The results of our investigations suggest that the main conditions for a reliable differential-screening experiment are fulfilled when using

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

    International Nuclear Information System (INIS)

    Kroczek, R.A.; Siebert, E.

    1990-01-01

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

  15. DISKRIMINASI KELAMIN PADA IKAN TUNA SIRIP KUNING, Yellowfin tuna MENGGUNAKAN ANALISIS DOT BLOT DAN ELISA

    Directory of Open Access Journals (Sweden)

    Gusti Ngurah Permana

    2014-08-01

    Full Text Available Pemahaman tentang penentuan jenis kelamin dalam populasi induk merupakan hal yang sangat penting bagi keberhasilan program pembenihan. Pengukuran reaksi antibodi dan aktivitas hormon testosterone, serta estradiol adalah metode dengan potensi yang secara akurat dapat menentukan jenis kelamin ikan tanpa mematikan ikan. Tujuan penelitian ini adalah untuk mengetahui akurasi metode dot blot dan ELISA dengan 11-ketotestorsterone (11-KT yang tersedia secara komersial EIA-kit untuk membedakan jenis kelamin ikan tuna sirip kuning. Hasil analisis menunjukkan bahwa metode dot blot menghasilkan ekspresi vitelogenin tampak jelas pada individu betina dan efek plasma terlihat transparan, jika dibandingkan dengan individu jantan. Interpretasi dari metode ini memerlukan pengalaman dan keahlian dalam akurasi pembacaan hasil. Aktivitas hormon 11-KT dengan sampel klip sirip dan plasma memberikan hasil yang baik dengan aktivitas hormon terlihat jelas.

  16. Reverse line blot probe design and polymerase chain reaction optimization for bloodmeal analysis of ticks from the eastern United States.

    Science.gov (United States)

    Scott, M C; Harmon, J R; Tsao, J I; Jones, C J; Hickling, G J

    2012-05-01

    Determining the host preference of vector ticks is vital to elucidating the eco-epidemiology of the diseases they spread. Detachment of ticks from captured hosts can provide evidence of feeding on those host species, but only for those species that are feasible to capture. Recently developed, highly sensitive molecular assays show great promise in allowing host selection to be determined from minute traces of host DNA that persist in recently molted ticks. Using methods developed in Europe as a starting-point, we designed 12S rDNA mitochondrial gene probes suitable for use in a reverse line blot (RLB) assay of ticks feeding on common host species in the eastern United States. This is the first study to use the 12S mitochondrial gene in a RLB bloodmeal assay in North America. The assay combines conventional PCR with a biotin-labeled primer and reverse line blots that can be stripped and rehybridized up to 20 times, making the method less expensive and more straightforward to interpret than previous methods of tick bloodmeal identification. Probes were designed that target the species, genus, genus group, family, order, or class of eight reptile, 13 birds, and 32 mammal hosts. After optimization, the RLB assay correctly identified the current hostspecies for 99% of ticks [Amblyomma americanum (L.) and eight other ixodid tick species] collected directly from known hosts. The method identified previous-host DNA for approximately half of all questing ticks assayed. Multiple bloodmeal determinations were obtained in some instances from feeding and questing ticks; this pattern is consistent with previous RLB studies but requires further investigation. Development of this probe library, suitable for eastern U.S. ecosystems, opens new avenues for eco-epidemiological investigations of this region's tick-host systems.

  17. Characterization of the structure of the erythropoietin receptor by ligand blotting

    International Nuclear Information System (INIS)

    Atkins, H.L.; Broudy, V.C.; Papayannopoulou, T.

    1991-01-01

    Erythropoietin (Epo) regulates the growth and differentiation of erythroid cells by binding to a specific receptor. We characterized the native Epo receptor on erythroleukemia cell lines by ligand blotting. Solubilized cell membrane proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred onto nitrocellulose, and probed with 125I-Epo. Specificity was demonstrated by inhibition of 125I-Epo binding by unlabeled excess Epo but not other peptide growth factors and by the cellular distribution of the Epo binding protein. A single membrane protein of 61 Kd ± 4 Kd was sufficient to bind 125I Epo in both human (OCIM2, K562) and murine (GM979, Rauscher, DA-1) cell lines. This finding is consistent with the predicted size of the Epo receptor from the murine cDNA clone. However, chemical crosslinking of 125I-Epo to its receptor has identified two Epo binding proteins of 105 Kd and 85 Kd. This difference may occur because the receptor is size fractionated before Epo binding in the ligand blot, but after Epo binding in crosslinking studies. Ligand blotting demonstrates that the native Epo receptor is composed of a single 61-Kd Epo binding protein, and suggests the presence of additional proteins of 20 to 25 Kd that associate with the receptor after Epo binding

  18. Inheritance of resistance to barley yellow dwarf virus detected by northern blot analysis

    International Nuclear Information System (INIS)

    Lorens, G.F.; Falk, B.W.; Qualset, C.O.

    1989-01-01

    Development of wheat (Triticum aestivum L.) cultivars tolerant to the barley yellow dwarf virus disease (BYD) has been limited by lack of precision in rating plants for response to infection, usually done by visual scoring of plant symptoms under field conditions. Other methodologies have been developed to study the host/pathogen relationship and to assess resistance or susceptibility. In this study northern dot blot analysis was used to determine barley yellow dwarf virus (BYDV) RNA concentrations of six wheat cultivars that differed in visual BYD symptom expression. Plants were infected with the NYPAV (PAV) isolate of BYDV in the greenhouse. At several dates after inoculation crude plant extracts were blotted on nitrocellulose and hybridized with a 32 P-labeled probe of the pPA8 cDNA clone of BYDV. The distribution of PRC for the F 2 population was compared to the distribution of BYD visual symptom scores for 403 F 2 plants of a similar F 2 population of NS 879/4 x Seri 82 under field conditions. The results were qualitatively similar, suggesting that northern dot blot analysis to measure PRC may be useful in understanding the genetics of resistance to BYD. This technique, when incorporated into breeding programs, could be important in the development of highly tolerant wheat cultivars with reduced losses to BYD

  19. Standardization of Licorice and TCM Formulations Using Eastern Blot Fingerprinting Analysis

    Directory of Open Access Journals (Sweden)

    Yukihiro Shoyama

    2013-01-01

    Full Text Available To prepare the antiglycyrrhizin (GC monoclonal antibody (MAb, GC was treated with NaIO4 resulting in aldehyde which can be combined with carrier protein. An antigen conjugate was performed by a matrix-assisted laser desorption/ionization TOF mass spectrometry to determine the hapten numbers in the conjugate. Anti-GC MAb was prepared from a hybridoma which was fixed from the spleen cells producing anti-GC MAb and the myeloma cells after immunization. The TCM and licorice extract were developed by TLC and blotted to a polyvinylidene difluoride (PVDF membrane. The membrane was treated by NaIO4 and protein, enzyme labeled secondary MAb, and finally substrate was added. Clear spot appeared on PVDF membrane identifying GC against a background containing large amount of impurities. In eastern blotting, the GC molecule was divided into two functions. The aglycone part is recognized as an epitope and the sugar moiety can be combined to membrane. The specific reactivity of sugar moiety in the GC molecule against anti-GC MAb might be modified by the NaIO4 treatment on the membrane because glycyrrhetic acid 3-O-glucuronide can be stained although the cross-reactivity is only 4.3%. Eastern blotting for GC can not only apply for the standardization of licorice and TCM, but also it can open for the other bioactive products.

  20. Dot-blot immunoassay of Fasciola gigantica infection using 27 kDa and adult worm regurge antigens in Egyptian patients.

    Science.gov (United States)

    Kamel, Hanan H; Saad, Ghada A; Sarhan, Rania M

    2013-04-01

    The purpose of the present study was to evaluate the potential role of the 27-Kilodalton (KDa) antigen versus Fasciola gigantica adult worm regurge antigens in a DOT-Blot assay and to assess this assay as a practical tool for diagnosis fascioliasis in Egyptian patients. Fasciola gigantica antigen of an approximate molecular mass 27-(KDa) was obtained from adult worms by a simple elution SDS-PAGE. A Dot-Blot was developed comparatively to adult worm regurge antigens for the detection of specific antibodies from patients infected with F. gigantica in Egypt. Control sera were obtained from patients with other parasitic infections and healthy volunteers to assess the test and compare between the antigens. The sensitivity, specificity, positive and negative predictive values of Dot-Blot using the adult worm regurge were 80%, 90%, 94.1%, and 69.2% respectively, while those using 27-KDa were 100% which confirms the diagnostic potential of this antigen. All patients infected with Fasciola were positive, with cross reactivity reported with Schistosoma mansoni serum samples. This 27-KDa Dot-Blot assay showed to be a promising test which can be used for serodiagnosis of fascioliasis in Egyptian patients especially, those presenting with hepatic disease. It is specific, sensitive and easy to perform method for the rapid diagnosis particularly when more complex laboratory tests are unavailable.

  1. Blot hybridization analysis of TCR genes of T cells for five people exposed in a radiation accident

    International Nuclear Information System (INIS)

    Min Rui; Liu Benti; Cheng Tianmin; Yang Rujun; Meng Xiangshun; Xiao Jinsong

    1996-01-01

    Human lymphocyte total DNA was prepared in agarose plug by mixing cells with low melting agarose, and two restriction endonucleases were used for digestion of the total DNA with human α and β TCR cDNA probes. The total digested DNA from five people who were whole body exposed to 2.0-2.5 Gy ionizing radiation in an accident 4.5 years ago was hybridized by Southern blot method. The results showed that no obvious difference in hybridization bands was found between controls and the five victims when hybridizations were fulfilled in the total DNA which was digested by Hind III restriction endonuclease with both α and β probes. However, when the total DNA was digested with restriction endonuclease EcoR I and was hybridized with TCR α probe, four of the five exposed people showed a different hybridizing band pattern compared with the controls. The results are also discussed

  2. A comparison of change detection measurements using object-based and pixel-based classification methods on western juniper dominated woodlands in eastern Oregon

    Directory of Open Access Journals (Sweden)

    Ryan G. Howell

    2017-03-01

    Full Text Available Encroachment of pinyon (Pinus spp and juniper (Juniperus spp. woodlands in western North America is considered detrimental due to its effects on ecohydrology, plant community structure, and soil stability. Management plans at the federal, state, and private level often include juniper removal for improving habitat of sensitive species and maintaining sustainable ecosystem processes. Remote sensing has become a useful tool in determining changes in juniper woodland structure because of its uses in comparing archived historic imagery with newly available multispectral images to provide information on changes that are no longer detectable by field measurements. Change in western juniper (J. occidentalis cover was detected following juniper removal treatments between 1995 and 2011 using panchromatic 1-meter NAIP and 4-band 1-meter NAIP imagery, respectively. Image classification was conducted using remotely sensed images taken at the Roaring Springs Ranch in southeastern Oregon. Feature Analyst for ArcGIS (object-based extraction and a supervised classification with ENVI 5.2 (pixel-based extraction were used to delineate juniper canopy cover. Image classification accuracy was calculated using an Accuracy Assessment and Kappa Statistic. Both methods showed approximately a 76% decrease in western juniper cover, although differing in total canopy cover area, with object-based classification being more accurate. Classification results for the 2011 imagery were much more accurate (0.99 Kappa statistic because of its low juniper density and the presence of an infrared band. The development of methods for detecting change in juniper cover can lead to more accurate and efficient data acquisition and subsequently improved land management and monitoring practices. These data can subsequently be used to assess and quantify juniper invasion and succession, potential ecological impacts, and plant community resilience.

  3. Projected changes over western Canada using convection-permitting regional climate model and the pseudo-global warming method

    Science.gov (United States)

    Li, Y.; Kurkute, S.; Chen, L.

    2017-12-01

    Results from the General Circulation Models (GCMs) suggest more frequent and more severe extreme rain events in a climate warmer than the present. However, current GCMs cannot accurately simulate extreme rainfall events of short duration due to their coarse model resolutions and parameterizations. This limitation makes it difficult to provide the detailed quantitative information for the development of regional adaptation and mitigation strategies. Dynamical downscaling using nested Regional Climate Models (RCMs) are able to capture key regional and local climate processes with an affordable computational cost. Recent studies have demonstrated that the downscaling of GCM results with weather-permitting mesoscale models, such as the pseudo-global warming (PGW) technique, could be a viable and economical approach of obtaining valuable climate change information on regional scales. We have conducted a regional climate 4-km Weather Research and Forecast Model (WRF) simulation with one domain covering the whole western Canada, for a historic run (2000-2015) and a 15-year future run to 2100 and beyond with the PGW forcing. The 4-km resolution allows direct use of microphysics and resolves the convection explicitly, thus providing very convincing spatial detail. With this high-resolution simulation, we are able to study the convective mechanisms, specifically the control of convections over the Prairies, the projected changes of rainfall regimes, and the shift of the convective mechanisms in a warming climate, which has never been examined before numerically at such large scale with such high resolution.

  4. Water quality assessment by pollution-index method in the coastal waters of Hebei Province in western Bohai Sea, China.

    Science.gov (United States)

    Liu, Shuguang; Lou, Sha; Kuang, Cuiping; Huang, Wenrui; Chen, Wujun; Zhang, Jianle; Zhong, Guihui

    2011-10-01

    Sources of pollution discharges and water quality samples at 27 stations in 2006 in the coastal waters of Hebei Province, western Bohai Sea, have been analyzed in this study. Pollutant loads from industrial sewages have shown stronger impact on the water environment than those from the general sewages. Analysis indicates that pollution of COD is mainly resulted from land-based point pollutant sources. For phosphate concentration, non-point source pollution from coastal ocean (fishing and harbor areas) plays an important role. To assess the water quality conditions, Organic Pollution Index and Eutrophication Index have been used to quantify the level of water pollution and eutrophication conditions. Results show that pollution was much heavier in the dry season than flood season in 2006. Based on COD and phosphate concentrations, results show that waters near Shahe River, Douhe River, Yanghe River, and Luanhe River were heavily polluted. Water quality in the Qinhuangdao area was better than those in the Tangshan and Cangzhou areas. Copyright © 2011 Elsevier Ltd. All rights reserved.

  5. Efficacy of Pitfall Trapping, Winkler and Berlese Extraction Methods for Measuring Ground-Dwelling Arthropods in Moist-Deciduous Forests in the Western Ghats

    Science.gov (United States)

    Sabu, Thomas K.; Shiju, Raj T.

    2010-01-01

    The present study provides data to decide on the most appropriate method for sampling of ground-dwelling arthropods measured in a moist-deciduous forest in the Western Ghats in South India. The abundance of ground-dwelling arthropods was compared among large numbers of samples obtained using pitfall trapping, Berlese and Winkler extraction methods. Highest abundance and frequency of most of the represented taxa indicated pitfall trapping as the ideal method for sampling of ground-dwelling arthropods. However, with possible bias towards surface-active taxa, pitfall-trapping data is inappropriate for quantitative studies, and Berlese extraction is the better alternative. Berlese extraction is the better method for quantitative measurements than the other two methods, whereas pitfall trapping would be appropriate for qualitative measurements. A comparison of the Berlese and Winkler extraction data shows that in a quantitative multigroup approach, Winkler extraction was inferior to Berlese extraction because the total number of arthropods caught was the lowest; and many of the taxa that were caught from an identical sample via Berlese extraction method were not caught. Significantly a greater frequency and higher abundance of arthropods belonging to Orthoptera, Blattaria, and Diptera occurred in pitfall-trapped samples and Psocoptera and Acariformes in Berlese-extracted samples than that were obtained in the other two methods, indicating that both methods are useful, one complementing the other, eliminating a chance for possible under-representation of taxa in quantitative studies. PMID:20673122

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

    Science.gov (United States)

    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.

  7. Secondary dentine as a sole parameter for age estimation: Comparison and reliability of qualitative and quantitative methods among North Western adult Indians

    Directory of Open Access Journals (Sweden)

    Jasbir Arora

    2016-06-01

    Full Text Available The indestructible nature of teeth against most of the environmental abuses makes its use in disaster victim identification (DVI. The present study has been undertaken to examine the reliability of Gustafson’s qualitative method and Kedici’s quantitative method of measuring secondary dentine for age estimation among North Western adult Indians. 196 (M = 85; F = 111 single rooted teeth were collected from the Department of Oral Health Sciences, PGIMER, Chandigarh. Ground sections were prepared and the amount of secondary dentine formed was scored qualitatively according to Gustafson’s (0–3 scoring system (method 1 and quantitatively following Kedici’s micrometric measurement method (method 2. Out of 196 teeth 180 samples (M = 80; F = 100 were found to be suitable for measuring secondary dentine following Kedici’s method. Absolute mean error of age was calculated by both methodologies. Results clearly showed that in pooled data, method 1 gave an error of ±10.4 years whereas method 2 exhibited an error of approximately ±13 years. A statistically significant difference was noted in absolute mean error of age between two methods of measuring secondary dentine for age estimation. Further, it was also revealed that teeth extracted for periodontal reasons severely decreased the accuracy of Kedici’s method however, the disease had no effect while estimating age by Gustafson’s method. No significant gender differences were noted in the absolute mean error of age by both methods which suggest that there is no need to separate data on the basis of gender.

  8. A rapid method to assess grape rust mites on leaves and observations from case studies in western Oregon vineyards

    Science.gov (United States)

    A rapid method for extracting eriophyoid mites was adapted from previous studies to provide growers and IPM consultants with a practical, efficient, and reliable tool to monitor for rust mites in vineyards. The rinse in bag (RIB) method allows quick extraction of mites from collected plant parts (sh...

  9. Improved Simplified Methods for Effective Seismic Analysis and Design of Isolated and Damped Bridges in Western and Eastern North America

    Science.gov (United States)

    Koval, Viacheslav

    The seismic design provisions of the CSA-S6 Canadian Highway Bridge Design Code and the AASHTO LRFD Seismic Bridge Design Specifications have been developed primarily based on historical earthquake events that have occurred along the west coast of North America. For the design of seismic isolation systems, these codes include simplified analysis and design methods. The appropriateness and range of application of these methods are investigated through extensive parametric nonlinear time history analyses in this thesis. It was found that there is a need to adjust existing design guidelines to better capture the expected nonlinear response of isolated bridges. For isolated bridges located in eastern North America, new damping coefficients are proposed. The applicability limits of the code-based simplified methods have been redefined to ensure that the modified method will lead to conservative results and that a wider range of seismically isolated bridges can be covered by this method. The possibility of further improving current simplified code methods was also examined. By transforming the quantity of allocated energy into a displacement contribution, an idealized analytical solution is proposed as a new simplified design method. This method realistically reflects the effects of ground-motion and system design parameters, including the effects of a drifted oscillation center. The proposed method is therefore more appropriate than current existing simplified methods and can be applicable to isolation systems exhibiting a wider range of properties. A multi-level-hazard performance matrix has been adopted by different seismic provisions worldwide and will be incorporated into the new edition of the Canadian CSA-S6-14 Bridge Design code. However, the combined effect and optimal use of isolation and supplemental damping devices in bridges have not been fully exploited yet to achieve enhanced performance under different levels of seismic hazard. A novel Dual-Level Seismic

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

    Science.gov (United States)

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

    2016-01-01

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

  11. Can cognitive dissonance methods developed in the West for combatting the 'thin ideal' help slow the rapidly increasing prevalence of eating disorders in non-Western cultures?

    Science.gov (United States)

    Witcomb, Gemma L; Arcelus, Jon; Chen, Jue

    2013-12-01

    Eating disorders are common, life-threatening conditions in Western countries, but until relatively recently they were regarded as uncommon in non-Western cultures. However, the prevalence of eating disorders in many of the more affluent non-Western countries is rising rapidly as community members, particularly young women, internalize the 'thin ideal' that has been widely promoted by the international media. This review discusses the factors involved in the development of eating disorders in non-Western settings with a particular emphasis on the influences of urbanization, modernization, Westernization, and the resulting changes in women's roles. The cognitive dissonance programs developed in Western countries that have proven successful in countering the negative effects of the thin idea are described and their potential application to East Asia and other non-Western countries are discussed.

  12. Evaluation of different storage methods to characterize the fecal bacterial communities of captive western lowland gorillas (Gorilla gorilla gorilla)

    Czech Academy of Sciences Publication Activity Database

    Vlčková, K.; Mrázek, Jakub; Kopečný, Jan; Petrželková, Klára Judita

    2012-01-01

    Roč. 91, č. 1 (2012), s. 45-51 ISSN 0167-7012 Institutional support: RVO:67985904 ; RVO:68081766 ; RVO:60077344 Keywords : storage methods * bacterial DNA * fecal sample Subject RIV: EE - Microbiology, Virology Impact factor: 2.161, year: 2012

  13. Estimating alcohol content of traditional brew in Western Kenya using culturally relevant methods: the case for cost over volume.

    Science.gov (United States)

    Papas, Rebecca K; Sidle, John E; Wamalwa, Emmanuel S; Okumu, Thomas O; Bryant, Kendall L; Goulet, Joseph L; Maisto, Stephen A; Braithwaite, R Scott; Justice, Amy C

    2010-08-01

    Traditional homemade brew is believed to represent the highest proportion of alcohol use in sub-Saharan Africa. In Eldoret, Kenya, two types of brew are common: chang'aa, spirits, and busaa, maize beer. Local residents refer to the amount of brew consumed by the amount of money spent, suggesting a culturally relevant estimation method. The purposes of this study were to analyze ethanol content of chang'aa and busaa; and to compare two methods of alcohol estimation: use by cost, and use by volume, the latter the current international standard. Laboratory results showed mean ethanol content was 34% (SD = 14%) for chang'aa and 4% (SD = 1%) for busaa. Standard drink unit equivalents for chang'aa and busaa, respectively, were 2 and 1.3 (US) and 3.5 and 2.3 (Great Britain). Using a computational approach, both methods demonstrated comparable results. We conclude that cost estimation of alcohol content is more culturally relevant and does not differ in accuracy from the international standard.

  14. Effect of a traditional processing method on the chemical composition of local white lupin (Lupinus albus L.) seed in North-Western Ethiopia.

    Science.gov (United States)

    Yeheyis, Likawent; Kijora, Claudia; Wink, Michael; Peters, Kurt J

    2011-01-01

    The effect of a traditional Ethiopian lupin processing method on the chemical composition of lupin seed samples was studied. Two sampling districts, namely Mecha and Sekela, representing the mid- and high-altitude areas of north-western Ethiopia, respectively, were randomly selected. Different types of traditionally processed and marketed lupin seed samples (raw, roasted, and finished) were collected in six replications from each district. Raw samples are unprocessed, and roasted samples are roasted using firewood. Finished samples are those ready for human consumption as snack. Thousand seed weight for raw and roasted samples within a study district was similar (P > 0.05), but it was lower (P seed sample from Mecha was lower (P 0.05). The crude protein and crude fat contents of finished samples within a study district were higher (P seed samples. The crude ash content of raw and roasted lupin samples within a study district was higher (P processing method of lupin seeds in Ethiopia has a positive contribution improving the crude protein and crude fat content, and lowering the alkaloid content of the finished product. The study showed the possibility of adopting the traditional processing method to process bitter white lupin for the use as protein supplement in livestock feed in Ethiopia, but further work has to be done on the processing method and animal evaluation.

  15. Evaluación de las pruebas dot blot y aglutinación de látex para el diagnóstico de cisticercosis en Perú

    Directory of Open Access Journals (Sweden)

    Eduardo Miranda-Ulloa

    Full Text Available Con el objetivo de evaluar las pruebas dot blot y aglutinación de látex para la detección de cisticercosis humana con antígeno de líquido de cisticerco de Taenia solium, se usaron 125 sueros humanos, de los cuales 60 procedían de personas con cisticercosis confirmada por Western Blot, 45 de personas con otras enfermedades parasitarias y 20 de personas aparentemente sanas. La concentración óptima del antígeno para impregnar las tiras dot blot fue de 0,01 ug/uL, y para impregnar las partículas de látex fue de 0,092 ug/uL. Para la prueba dot blot se encontró una sensibilidad del 100% y especificidad del 87,7%; para la aglutinación de látex una sensibilidad del 93,3% y especificidad del 89,2%. Ambas pruebas podrían ser de utilidad y factibles de implementar como alternativas de diagnóstico serológico en laboratorios de áreas endémicas del Perú

  16. Molecular evidence for the occurrence of beet western yellows virus on chickpea in Morocco.

    NARCIS (Netherlands)

    Fortass, M.; Wilk, van der F.; Heuvel, van de J.F.J.M.; Goldbach, R.W.

    1997-01-01

    A luteovirus isolate infecting chickpea in Morocco was experimentally transmitted by Myzus persicae to Physalis floridana, on which it produced mild symptoms. When tested in western blots against antisera to known legume luteoviruses, this isolate reacted strongly to beet western yellows virus

  17. Determinants of long acting and permanent contraceptive methods utilization among married women of reproductive age groups in western Ethiopia: a cross-sectional study.

    Science.gov (United States)

    Melka, Alemu Sufa; Tekelab, Tesfalidet; Wirtu, Desalegn

    2015-01-01

    In Ethiopia information on the level of utilization of the long term and permanent contraceptive methods and associated factorsis lacking. The aim of this study was to understand the determinant factors of long acting and permanent contraceptive methods use among married women of reproductive age in Western Ethiopia. A community based cross-sectional study design was employed. Multi stage sampling was used to select 1003 study participants. Data was collected from April 10 to April 25,2014 using a pre- tested structured questionnaire. The data were entered using Epi-info version 3.5.1 and exported to SPSS version 20 for analysis. Multivariate logistic regression analysis was done to identify predictors of long acting and permanent contraceptive methods at 95% CL. Use of long acting and permanent contraceptive methods in this study was found to be 20%. Survey results showed a significant positive association between utilization of long acting and permanent contraceptive methods and women's education (AOR=1.72, 95%CI=1.02-3.05), women's occupation (AOR=2.01, 95% CI=1.11-3.58), number of live children (AOR=2.42, 95% CI: 1.46-4.02), joint fertility related decision (AOR=6.11, 95% CI: 2.29-16.30), having radio/TV (AOR=2.31, 95% CI: 1.40-3.80), and discussion with health care provider about long acting and permanent contraceptive methods (AOR=13.72, 95% CI: 8.37-22.47). Efforts need to be aimed at women empowerment, health education, and encouraging open discussion of family planning by couples.

  18. Comparison of RT-PCR-Dot blot hybridization based on radioisotope 32P with conventional RT-PCR and commercial ELISA Assays for blood screening of HIV-1

    International Nuclear Information System (INIS)

    Maria Lina R; Andi Yasmon

    2011-01-01

    There are many commercial ELISA and rapid test kits that have been used for blood screening; however, the kits can give false positive and negative results. Therefore, RT-PCR (Reverse Transcription Polymerase Chain Reaction) - Dot Blot Hybridization based on radioisotope 32 P (RDBR) method was developed in this research, to compare the method with the conventional RT-PCR and commercial ELISA Enzyme-Linked lmmunosorbent Assay) kit. This method is efficient for screening of large blood specimens and surveillance study. Eighty seven samples were used and serum of the samples were tested by ELISA to detect HIV-1. The HIV-l RNA genome was extracted from plasma samples and tested using the RT-PCR and RDBR methods. Of 87 samples that were tested, the rates of positive testing of the RT-PCR, the RDBR, and the ELISA were 71.26%, 74.71%, and 80.46%, respectively. The RDBR (a combination of RTPCR and dot blot hybridization) was more sensitive than conventional RT-PCR by showing 3.45% in increase number of positive specimens. The results showed that of 9 samples (10.34%) were negative RDBR and positive ELISA, while 4 samples (4.60%) were negative ELISA and positive RDBR. The two methods showed slightly difference in the results but further validation is still needed. However, RDBR has high potential as an alternative method for screening of blood in large quantities when compared to method of conventional RT-PCR and ELISA. (author)

  19. A laser ablation ICP-MS based method for multiplexed immunoblot analysis

    DEFF Research Database (Denmark)

    de Bang, Thomas Christian; Petersen, Jørgen; Pedas, Pai Rosager

    2015-01-01

    developed a multiplexed antibody-based assay and analysed selected PSII subunits in barley (Hordeum vulgare L.). A selection of antibodies were labelled with specific lanthanides and immunoreacted with thylakoids exposed to Mn deficiency after western blotting. Subsequently, western blot membranes were...... analysed by laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS), which allowed selective and relative quantitative analysis via the different lanthanides. The method was evaluated against established liquid chromatography electrospray ionization tandem mass spectrometry (LC...... by more than one technique. The developed method enables a higher number of proteins to be multiplexed in comparison to existing immunoassays. Furthermore, multiplexed protein analysis by LA-ICP-MS provides an analytical platform with high throughput appropriate for screening large collections of plants....

  20. Rosette Assay: Highly Customizable Dot-Blot for SH2 Domain Screening.

    Science.gov (United States)

    Ng, Khong Y; Machida, Kazuya

    2017-01-01

    With a growing number of high-throughput studies, structural analyses, and availability of protein-protein interaction databases, it is now possible to apply web-based prediction tools to SH2 domain-interactions. However, in silico prediction is not always reliable and requires experimental validation. Rosette assay is a dot blot-based reverse-phase assay developed for the assessment of binding between SH2 domains and their ligands. It is conveniently customizable, allowing for low- to high-throughput analysis of interactions between various numbers of SH2 domains and their ligands, e.g., short peptides, purified proteins, and cell lysates. The binding assay is performed in a 96-well plate (MBA or MWA apparatus) in which a sample spotted membrane is incubated with up to 96 labeled SH2 domains. Bound domains are detected and quantified using a chemiluminescence or near-infrared fluorescence (IR) imaging system. In this chapter, we describe a practical protocol for rosette assay to assess interactions between synthesized tyrosine phosphorylated peptides and a library of GST-tagged SH2 domains. Since the methodology is not confined to assessment of SH2-pTyr interactions, rosette assay can be broadly utilized for ligand and drug screening using different protein interaction domains or antibodies.

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

    Science.gov (United States)

    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.

  2. Direct tissue blot immunoassay for detection of Xylella fastidiosa in olive trees

    Directory of Open Access Journals (Sweden)

    Khaled DJELOUAH

    2015-01-01

    Full Text Available A direct tissue blot immunoassay (DTBIA technique has been compared with ELISA and PCR for detection of Xylella fastidiosa in olive trees from Apulia (southern Italy. Fresh cross-sections of young twigs and leaf petioles were printed onto nitrocellulose membranes and analyzed in the laboratory. Analyses of a first group of 61 samples gave similar efficiency for the three diagnostic techniques for detection the bacterium (24 positive and 36 negative samples, except for a single sample which was positive only with DTBIA and PCR. Similar results were obtained by separately analyzing suckers and twigs collected from different sectors of tree canopies of a second group of 20 olive trees (ten symptomatic and ten symptomless. In this second test the three diagnostic techniques confirmed the irregular distribution of the bacterium in the tree canopies and erratic detectability of the pathogen in the young suckers. It is therefore necessary to analyse composite samples per tree which should be prepared with twigs collected from different sides of the canopy. The efficiency comparable to ELISA and PCR, combined with the advantages of easier handling, speed and cost, make DTBIA a valid alternative to ELISA in large-scale surveys for occurrence of X. fastidiosa. Moreover, the printing of membranes directly in the field prevents infections spreading to Xylella-free areas, through movement of plant material with pathogen vectors for laboratory testing.

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

    Directory of Open Access Journals (Sweden)

    William H Roldán

    2009-05-01

    Full Text Available 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.

  4. Retrospective study of hemoparasites in cattle in southern Italy by reverse line blot hybridization.

    Science.gov (United States)

    Ceci, Luigi; Iarussi, Fabrizio; Greco, Beatrice; Lacinio, Rosanna; Fornelli, Stefania; Carelli, Grazia

    2014-06-01

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

  5. Simultaneous Detection of Bovine Theileria and Babesia Species by Reverse Line Blot Hybridization

    Science.gov (United States)

    Gubbels, J. M.; de Vos, A. P.; van der Weide, M.; Viseras, J.; Schouls, L. M.; de Vries, E.; Jongejan, F.

    1999-01-01

    A reverse line blot (RLB) assay was developed for the identification of cattle carrying different species of Theileria and Babesia simultaneously. We included Theileria annulata, T. parva, T. mutans, T. taurotragi, and T. velifera in the assay, as well as parasites belonging to the T. sergenti-T. buffeli-T. orientalis group. The Babesia species included were Babesia bovis, B. bigemina, and B. divergens. The assay employs one set of primers for specific amplification of the rRNA gene V4 hypervariable regions of all Theileria and Babesia species. PCR products obtained from blood samples were hybridized to a membrane onto which nine species-specific oligonucleotides were covalently linked. Cross-reactions were not observed between any of the tested species. No DNA sequences from Bos taurus or other hemoparasites (Trypanosoma species, Cowdria ruminantium, Anaplasma marginale, and Ehrlichia species) were amplified. The sensitivity of the assay was determined at 0.000001% parasitemia, enabling detection of the carrier state of most parasites. Mixed DNAs from five different parasites were correctly identified. Moreover, blood samples from cattle experimentally infected with two different parasites reacted only with the corresponding species-specific oligonucleotides. Finally, RLB was used to screen blood samples collected from carrier cattle in two regions of Spain. T. annulata, T. orientalis, and B. bigemina were identified in these samples. In conclusion, the RLB is a versatile technique for simultaneous detection of all bovine tick-borne protozoan parasites. We recommend its use for integrated epidemiological monitoring of tick-borne disease, since RLB can also be used for screening ticks and can easily be expanded to include additional hemoparasite species. PMID:10325324

  6. Epidemiology of Babesia, Anaplasma and Trypanosoma species using a new expanded reverse line blot hybridization assay.

    Science.gov (United States)

    Paoletta, Martina Soledad; López Arias, Ludmila; de la Fournière, Sofía; Guillemi, Eliana Carolina; Luciani, Carlos; Sarmiento, Néstor Fabián; Mosqueda, Juan; Farber, Marisa Diana; Wilkowsky, Silvina Elizabeth

    2018-02-01

    Vector-borne hemoparasitic infections are a major problem that affects livestock industries worldwide, particularly in tropical and subtropical regions. In this work, a reverse line blot (RLB) hybridization assay was developed for the simultaneous detection and identification of Anaplasma, Babesia and bovine trypanosomes, encompassing in this way the most relevant hemoparasites that affect cattle. A total of 186 bovine blood samples collected from two different ecoepidemiological regions of northeast Argentina, with and without tick control, were analyzed with this new RLB. High diversity of parasites, such as Babesia bovis, B. bigemina, Anaplasma marginale and three different Trypanosoma species, was found. High rates of coinfections were also detected, and significant differences were observed not only in the prevalence of parasites but also in the level of coinfections between the two analyzed areas. Regarding the Trypanosoma genus, we provide molecular evidence of the presence of T. vivax and T. theileri for the first time in Argentina. Besides, since the RLB is a prospective tool, it allowed the identification of a yet unknown bovine trypanosome which could not be assigned to any of the bovine species known so far. In the present study we provide new insights on the prevalence of several pathogens that directly impact on livestock production in Argentina. The RLB assay developed here allows to identify simultaneously numerous pathogenic species which can also be easily expanded to detect other blood borne pathogens. These characteristics make the RLB hybridization assay an essential tool for epidemiological survey of all vector-borne pathogens. Copyright © 2017 Elsevier GmbH. All rights reserved.

  7. Predictors of modern contraceptive methods use among married women of reproductive age groups in Western Ethiopia: a community based cross-sectional study.

    Science.gov (United States)

    Tekelab, Tesfalidet; Melka, Alemu Sufa; Wirtu, Desalegn

    2015-07-17

    In Ethiopia, the prevalence of modern contraceptive use is very low (27 %) and the percentage of those with unmet needs for family planning is 25 %. The current study identified factors associated with the utilization of modern contraceptive methods among married women in Western Ethiopia. A community based, cross-sectional study was employed from April 10 to April 25, 2014, among married women of reproductive age in Nekemte Town. A multi-stage sampling procedure was used to select 1003 study participants. A pretested structured questionnaire was used to collect data, and data collectors who had completed high school were involved in the data collection process. A bivariate, multivariable logistic regression model was fit, and statistical significance was determined with a 95% confidence level. The overall utilization rate of modern contraceptives in this study was 71.9%. The most common form of modern contraceptives used was injectable (60.3%). Age (AOR = 2.00, 95 % CI = 1.35-2.98), women's educational level (AOR = 2.50, 95 % CI = 1.62-3.84), monthly income (AOR = 2.26, 95 % CI = 1.24-4.10), respondent's fertility (AOR = 2.60, 95 % CI = 1.48-4.56), fertility-related decision (AOR = 3.70, 95 % CI = 2.45-5.58), and having radio (AOR = 1.93, 95 % CI = 1.37-2.71) showed significant positive associations with the utilization of modern contraceptive methods. The findings showed that women's empowerment, fertility-related discussions among couples, and the availability of the media were important factors that influenced the use of modern contraceptives. Thus, policymakers and implementers should work on those factors to increase the utilization of modern contraceptive methods.

  8. Sulphate reduction and vertical distribution of sulphate-reducing bacteria quantified by rRNA slot-blot hybridization in a coastal marine sediment

    DEFF Research Database (Denmark)

    Sahm, K.; MacGregor, BJ; Jørgensen, BB

    1999-01-01

    In the past, enumeration of sulphate-reducing bacteria (SRB) by cultivation-based methods generally contradicted measurements of sulphate reduction, suggesting unrealistically high respiration rates per cell. Here, we report evidence that quantification of SRB rRNA by slot-blot hybridization......, directly above the sulphate reduction maximum. Cell numbers calculated by converting the relative contribution of SRB rRNA to the percentage of DAPI-stained cells indicated a population size for SRB of 2.4-6.1 x 10(8) cells cm(-3) wet sediment. Cellular sulphate reduction rates calculated on the basis...

  9. Comparison of fumonisin contamination using HPLC and ELISA methods in bt and near-isogenic maize hybrids infested with European corn borer or western bean cutworm.

    Science.gov (United States)

    Bowers, Erin; Hellmich, Richard; Munkvold, Gary

    2014-07-09

    Field trials were conducted from 2007 to 2010 to compare grain fumonisin levels among non-Bt maize hybrids and Bt hybrids with transgenic protection against manual infestations of European corn borer (ECB) and Western bean cutworm (WBC). HPLC and ELISA were used to measure fumonisin levels. Results of the methods were highly correlated, but ELISA estimates were higher. Bt hybrids experienced less insect injury, Fusarium ear rot, and fumonisin contamination compared to non-Bt hybrids. WBC infestation increased fumonisin content compared to natural infestation in non-Bt and hybrids expressing Cry1Ab protein in five of eight possible comparisons; in Cry1F hybrids, WBC did not impact fumonisins. These results indicate that WBC is capable of increasing fumonisin levels in maize. Under WBC infestation, Cry1F mitigated this risk more consistently than Cry1Ab or non-Bt hybrids. Transgenically expressed Bt proteins active against multiple lepidopteran pests can provide broad, consistent reductions in the risk of fumonisin contamination.

  10. An appraisal of the geologic structure beneath the Ikogosi warm spring in south-western Nigeria using integrated surface geophysical methods

    Directory of Open Access Journals (Sweden)

    J.S Ojo

    2011-06-01

    Full Text Available An integrated surface geophysical investigation involving resistivity and magnetic methods was carried out in the immediate vicinity of the Ikogosi warm spring situated in south-western Nigeria with a view to delineating its subsurface geological sequence and evaluating the structural setting beneath the warmspring. Total field magnetic measurements and vertical electrical sounding (VES data were acquired along five N-S traverses. Magnetic and VES data interpretation
    involved inverse modelling. The inverse magnetic models delineated fractured quartzite/faulted areas within fresh massive quartzite at varying depths and beneath all traverses. The geoelectrical sections developed from VESinterpretation results also delineated a subsurface sequence consisting of a topsoil/weathered layer, fresh quartzite, fractured/faulted quartzite and fresh quartzite bedrock. It was deduced that the fractured/faulted quartzite may have acted as conduit for the
    movement of warm groundwater from profound depths to the surface while the spring outlet was located on a geological interface  (lineament.

  11. Patterns of Limnohabitans Microdiversity across a Large Set of Freshwater Habitats as Revealed by Reverse Line Blot Hybridization

    Science.gov (United States)

    Jezbera, Jan; Jezberová, Jitka; Kasalický, Vojtěch; Šimek, Karel; Hahn, Martin W.

    2013-01-01

    Among abundant freshwater Betaproteobacteria, only few groups are considered to be of central ecological importance. One of them is the well-studied genus Limnohabitans and mainly its R-BT subcluster, investigated previously mainly by fluorescence in situ hybridization methods. We designed, based on sequences from a large Limnohabitans culture collection, 18 RLBH (Reverse Line Blot Hybridization) probes specific for different groups within the genus Limnohabitans by targeting diagnostic sequences on their 16 S–23 S rRNA ITS regions. The developed probes covered in sum 92% of the available isolates. This set of probes was applied to environmental DNA originating from 161 different European standing freshwater habitats to reveal the microdiversity (intra-genus) patterns of the Limnohabitans genus along a pH gradient. Investigated habitats differed in various physicochemical parameters, and represented a very broad range of standing freshwater habitats. The Limnohabitans microdiversity, assessed as number of RLBH-defined groups detected, increased significantly along the gradient of rising pH of habitats. 14 out of 18 probes returned detection signals that allowed predictions on the distribution of distinct Limnohabitans groups. Most probe-defined Limnohabitans groups showed preferences for alkaline habitats, one for acidic, and some seemed to lack preferences. Complete niche-separation was indicated for some of the probe-targeted groups. Moreover, bimodal distributions observed for some groups of Limnohabitans, suggested further niche separation between genotypes within the same probe-defined group. Statistical analyses suggested that different environmental parameters such as pH, conductivity, oxygen and altitude influenced the distribution of distinct groups. The results of our study do not support the hypothesis that the wide ecological distribution of Limnohabitans bacteria in standing freshwater habitats results from generalist adaptations of these bacteria

  12. Patterns of Limnohabitans microdiversity across a large set of freshwater habitats as revealed by Reverse Line Blot Hybridization.

    Directory of Open Access Journals (Sweden)

    Jan Jezbera

    Full Text Available Among abundant freshwater Betaproteobacteria, only few groups are considered to be of central ecological importance. One of them is the well-studied genus Limnohabitans and mainly its R-BT subcluster, investigated previously mainly by fluorescence in situ hybridization methods. We designed, based on sequences from a large Limnohabitans culture collection, 18 RLBH (Reverse Line Blot Hybridization probes specific for different groups within the genus Limnohabitans by targeting diagnostic sequences on their 16 S-23 S rRNA ITS regions. The developed probes covered in sum 92% of the available isolates. This set of probes was applied to environmental DNA originating from 161 different European standing freshwater habitats to reveal the microdiversity (intra-genus patterns of the Limnohabitans genus along a pH gradient. Investigated habitats differed in various physicochemical parameters, and represented a very broad range of standing freshwater habitats. The Limnohabitans microdiversity, assessed as number of RLBH-defined groups detected, increased significantly along the gradient of rising pH of habitats. 14 out of 18 probes returned detection signals that allowed predictions on the distribution of distinct Limnohabitans groups. Most probe-defined Limnohabitans groups showed preferences for alkaline habitats, one for acidic, and some seemed to lack preferences. Complete niche-separation was indicated for some of the probe-targeted groups. Moreover, bimodal distributions observed for some groups of Limnohabitans, suggested further niche separation between genotypes within the same probe-defined group. Statistical analyses suggested that different environmental parameters such as pH, conductivity, oxygen and altitude influenced the distribution of distinct groups. The results of our study do not support the hypothesis that the wide ecological distribution of Limnohabitans bacteria in standing freshwater habitats results from generalist adaptations of

  13. A specific and sensitive method for visualization of tumor necrosis factor in the murine central nervous system

    DEFF Research Database (Denmark)

    Lambertsen, K L; Drøjdahl, N; Owens, T

    2001-01-01

    -PCR and Western blot analysis on homogenates prepared from microdissected brain regions. Advantages and disadvantages of the methods are discussed with emphasis on the specificity and sensitivity of the histological procedures. Our strategy for detection of TNF mRNA and protein provides a solid basis...... for clarifying the cellular synthesis, regulation and function of TNF in the normal, injured or diseased CNS. Furthermore, the methodology can readily be applied in studies of other cytokines and growth factors in the CNS....

  14. Different domains of Bacillus thuringiensis delta-endotoxins can bind to insect midgut membrane proteins on ligand blots

    NARCIS (Netherlands)

    Maagd, de R.A.; Klei, van der H.; Bakker, P.L.; Stiekema, W.J.; Bosch, D.

    1996-01-01

    We investigated the role of the constituent domains of the CryIA(b) and CryIA(c) δ-endotoxins in binding to midgut epithelial cell membrane proteins of Spodoptera exigua and Manduca sexta on ligand blots. A collection of wild- type and CryIC-CryIA hybrid toxins was used for this purpose. As

  15. Diagnosing cancer in the bush: a mixed-methods study of symptom appraisal and help-seeking behaviour in people with cancer from rural Western Australia.

    Science.gov (United States)

    Emery, Jon D; Walter, Fiona M; Gray, Vicky; Sinclair, Craig; Howting, Denise; Bulsara, Max; Bulsara, Caroline; Webster, Andrew; Auret, Kirsten; Saunders, Christobel; Nowak, Anna; Holman, C D'Arcy

    2013-06-01

    Previous studies have focused on the treatment received by rural cancer patients and have not examined their diagnostic pathways as reasons for poorer outcomes in rural Australia. To compare and explore symptom appraisal and help-seeking behaviour in patients with breast, lung, prostate or colorectal cancer from rural Western Australia (WA). A mixed-methods study of people recently diagnosed with breast, lung, prostate or colorectal cancer from rural WA. The time from first symptom to diagnosis (i.e. total diagnostic interval, TDI) was calculated from interviews and medical records. Sixty-six participants were recruited (24 breast, 20 colorectal, 14 prostate and 8 lung cancer patients). There was a highly significant difference in time from symptom onset to seeking help between cancers (P = 0.006). Geometric mean symptom appraisal for colorectal cancer was significantly longer than that for breast and lung cancers [geometric mean differences: 2.58 (95% confidence interval, CI: 0.64-4.53), P = 0.01; 3.97 (1.63-6.30), P = 0.001, respectively]. There was a significant overall difference in arithmetic mean TDI (P = 0.046); breast cancer TDI was significantly shorter than colorectal or prostate cancer TDI [mean difference : 266.3 days (95% CI: 45.9-486.8), P = 0.019; 277.0 days, (32.1-521.9), P = 0.027, respectively]. These differences were explained by the nature and personal interpretation of symptoms, perceived as well as real problems of access to health care, optimism, stoicism, machismo, fear, embarrassment and competing demands. Longer symptom appraisal was observed for colorectal cancer. Participants defined core characteristics of rural Australians as optimism, stoicism and machismo. These features, as well as access to health care, contribute to later presentation of cancer.

  16. Blending Aboriginal and Western healing methods to treat intergenerational trauma with substance use disorder in Aboriginal peoples who live in northeastern Ontario, Canada.

    Science.gov (United States)

    Marsh, Teresa Naseba; Coholic, Diana; Cote-Meek, Sheila; Najavits, Lisa M

    2015-05-20

    As with many Indigenous groups around the world, Aboriginal communities in Canada face significant challenges with trauma and substance use. The complexity of symptoms that accompany intergenerational trauma and substance use disorders represents major challenges in the treatment of both disorders. There appears to be an underutilization of substance use and mental health services, substantial client dropout rates, and an increase in HIV infections in Aboriginal communities in Canada. The aim of this paper is to explore and evaluate current literature on how traditional Aboriginal healing methods and the Western treatment model "Seeking Safety" could be blended to help Aboriginal peoples heal from intergenerational trauma and substance use disorders. A literature search was conducted using the keywords: intergenerational trauma, historical trauma, Seeking Safety, substance use, Two-Eyed Seeing, Aboriginal spirituality, and Aboriginal traditional healing. Through a literature review of Indigenous knowledge, most Indigenous scholars proposed that the wellness of an Aboriginal community can only be adequately measured from within an Indigenous knowledge framework that is holistic, inclusive, and respectful of the balance between the spiritual, emotional, physical, and social realms of life. Their findings indicate that treatment interventions must honour the historical context and history of Indigenous peoples. Furthermore, there appears to be strong evidence that strengthening cultural identity, community integration, and political empowerment can enhance and improve mental health and substance use disorders in Aboriginal populations. In addition, Seeking Safety was highlighted as a well-studied model with most populations, resulting in healing. The provided recommendations seek to improve the treatment and healing of Aboriginal peoples presenting with intergenerational trauma and addiction. Other recommendations include the input of qualitative and quantitative

  17. A comparison of survey methods for documenting presence of Myotis leibii (Eastern Small-Footed Bats) at roosting areas in Western Virginia

    Science.gov (United States)

    Huth, John K.; Silvis, Alexander; Moosman, Paul R.; Ford, W. Mark; Sweeten, Sara E.

    2015-01-01

    Many aspects of foraging and roosting habitat of Myotis leibii (Eastern Small-Footed Bat), an emergent rock roosting-obligate, are poorly described. Previous comparisons of effectiveness of acoustic sampling and mist-net captures have not included Eastern Small-Footed Bat. Habitat requirements of this species differ from congeners in the region, and it is unclear whether survey protocols developed for other species are applicable. Using data from three overlapping studies at two sampling sites in western Virginia’s central Appalachian Mountains, detection probabilities were examined for three survey methods (acoustic surveys with automated identification of calls, visual searches of rock crevices, and mist-netting) for use in the development of “best practices” for future surveys and monitoring. Observer effects were investigated using an expanded version of visual search data. Results suggested that acoustic surveys with automated call identification are not effective for documenting presence of Eastern Small-Footed Bats on talus slopes (basal detection rate of 0%) even when the species is known to be present. The broadband, high frequency echolocation calls emitted by Eastern Small-Footed Bat may be prone to attenuation by virtue of their high frequencies, and these factors, along with signal reflection, lower echolocation rates or possible misidentification to other bat species over talus slopes may all have contributed to poor acoustic survey success. Visual searches and mist-netting of emergent rock had basal detection probabilities of 91% and 75%, respectively. Success of visual searches varied among observers, but detection probability improved with practice. Additionally, visual searches were considerably more economical than mist-netting.

  18. Comparison of self-report and interview administration methods based on the Brazilian versions of the Western Ontario Rotator Cuff Index and Disabilities of the Arm, Shoulder and Hand Questionnaire in patients with rotator cuff disorders

    Directory of Open Access Journals (Sweden)

    Andréa Diniz Lopes

    2009-02-01

    Full Text Available OBJECTIVE: The purpose of the present study was to compare self-report and interview administration methods using the Western Ontario Rotator Cuff Index (WORC and Disabilities of the Arm, Shoulder and Hand Questionnaire (DASH in patients with rotator cuff disorders. METHODS: Thirty male and female patients over 18 years of age with rotator cuff disorders (tendinopathy or rotator cuff tear and Brazilian Portuguese as their primary language were recruited for assessment via administration of the Western Ontario Rotator Cuff Index and and Disabilities of the Arm, Shoulder and Hand Questionnaire. A randomization method was used to determine whether the questionnaires would be self-reported (n=15 or administered by an interviewer (n=15. Pearson correlation coefficients were used to evaluate the correlation between the Western Ontario Rotator Cuff Index and and Disabilities of the Arm, Shoulder and Hand Questionnaire in each group. The t-test was used to determine whether the difference in mean questionnaire scores and administration time was statistically significant. For statistical analysis, the level of significance was set at 5%. RESULTS: The mean subject age was 55.07 years, ranging from 27 to 74 years. Most patients had a diagnosis of tendinopathy (n=21. With regard to level of schooling, the majority (n=26 of subjects had completed a college degree or higher. The mean questionnaire scores and administration times did not significantly differ between the two groups (p>0.05. There were statistically significant correlations (p<0.05 between Western Ontario Rotator Cuff Index and and Disabilities of the Arm, Shoulder and Hand Questionnaire, and strong correlations were found between the questionnaires in both groups. CONCLUSION: There are no differences between the Western Ontario Rotator Cuff Index and Disabilities of the Arm, Shoulder and Hand Questionnaire administration methods with regard to administration time or correlations between the

  19. Assessment of high-risk human papillomavirus infections using clinician- and self-collected cervical sampling methods in rural women from far western Nepal.

    Directory of Open Access Journals (Sweden)

    Derek C Johnson

    Full Text Available Nepal has one of the highest cervical cancer rates in South Asia. Only a few studies in populations from urban areas have investigated type specific distribution of human papillomavirus (HPV in Nepali women. Data on high-risk HPV (HR-HPV types are not currently available for rural populations in Nepal. We aimed to assess the distribution of HR- HPV among rural Nepali women while assessing self-collected and clinician-collected cervico-vaginal specimens as sample collection methods for HPV screening.Study participants were recruited during a health camp conducted by Nepal Fertility Care Center in Achham District of rural far western Nepal. Women of reproductive age completed a socio-demographic and clinical questionnaire, and provided two specimens; one cervical-vaginal specimen using a self-collection method and another cervical specimen collected by health camp auxiliary nurse midwives during a pelvic examination. All samples were tested for 14 different HR-HPV mRNA and also specific for HPV16/18/45 mRNA.Of 261 women with both clinician- and self-collected cervical samples, 25 tested positive for HR-HPV, resulting in an overall HR-HPV prevalence of 9.6% (95% confidence Interval [CI]: 6.3-13.8. The overall Kappa value assessing agreement between clinician- and self-collected tests was 0.62 (95% CI: 0.43-0.81, indicating a "good" level of agreement. Abnormal cytology was reported for 8 women. One woman identified with squamous cell carcinoma (SCC, and 7 women with high grade squamous intraepithelial lesions (HSIL. Seven of the 8 women tested positive for HR-HPV (87.5% in clinician-collected samples and 6 in self-collected samples (75.0%.This is the first study to assess HR-HPV among rural Nepali women. Self-collected sampling methods should be the subject of additional research in Nepal for screening HR-HPV, associated with pre-cancer lesions and cancer, in women in rural areas with limited access to health services.

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

    NARCIS (Netherlands)

    Thiry, D.; Mauroy, A.; Saegerman, C.; Thomas, I.; Wautier, M.; Miry, C.; Czaplicki, G.; Berkvens, D.; Praet, N.; Poel, van der W.H.M.; Cariolet, R.; Brochier, B.; Thiry, E.

    2014-01-01

    Zoonotic transmission of hepatitis E virus (HEV) is of special concern, particularly in high income countries were waterborne infections are less frequent than in developing countries. High HEV seroprevalences can be found in European pig populations. The aims of this study were to obtain prevalence

  1. A Western Blot-based Investigation of the Yeast Secretory Pathway Designed for an Intermediate-Level Undergraduate Cell Biology Laboratory

    Science.gov (United States)

    Hood-DeGrenier, Jennifer K.

    2008-01-01

    The movement of newly synthesized proteins through the endomembrane system of eukaryotic cells, often referred to generally as the secretory pathway, is a topic covered in most intermediate-level undergraduate cell biology courses. An article previously published in this journal described a laboratory exercise in which yeast mutants defective in…

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

    Science.gov (United States)

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

    2013-06-26

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

  3. Study on sensitivity of southern blotting hybridization using a 32P-labeled probe of PCR products in detecting human cytomegalovirus

    International Nuclear Information System (INIS)

    Bu Hengfu; Chen Juan; Shen Rongsen; Ma Liren; Xu Yongqiang

    1996-01-01

    Southern blotting hybridization (SBH) using a 32 P-labeled probe is one of the most practical methods for genetic diagnosis of pathogen. On the basis of establishing PCR and nested PCR for detecting human cytomegalovirus (HCMV), a 32 P-labeled probe was prepared with the amplified products of 613 bp PCR outer primers and hybridized with 300 bp inner primer amplified product, resulting in increase in detecting sensitivity from 17 ng (in 1.2% agarose electrophoresis) before SBH to 500 pg (autoradiographed), in other words, increasing the sensitivity of detecting HCMV by 10 2 dilutions after using SBH. The method of PCR and SBH using a 32 P-labeled probe could detect less than 1 gene copy of HCMV, therefore, it is a rapid and reliable diagnosis method for detecting HCMV latent infection

  4. A 15-year-long Southern blotting analysis of FMR1 to detect female carriers and for prenatal diagnosis of fragile X syndrome in Taiwan.

    Science.gov (United States)

    Tzeng, C-C; Tsai, L-P; Chang, Y-K; Hung, Y-J; Chang, Y-Y; Su, Y-P; Jiang, J-J; Liang, H-M

    2017-08-01

    Here, we review the results of Southern blotting analyses of the FMR1 gene performed in our reference laboratory in Taiwan over a 15-year period. In total, 725 high-risk women with a family history of fragile X syndrome (FXS) or idiopathic intellectual disability, 3911 low-risk pregnant women without such family history, and prenatal diagnosis data for 32 foetuses from 24 carrier mothers were included. Only 2 carriers were in the low-risk group, which indicated a prevalence of 1 of 1955 women (95% confidence interval: 1/7156-1/539). A total of 100 carriers were found to be in the high-risk group, thus revealing a significantly higher frequency than the low-risk group (100/725 vs 2/3911, P<0.0001). Eight of the 14 foetuses that inherited the maternal mutant allele were verified to have a full mutation, with the smallest maternal pre-mutation allele carrying 56 CGG repeats. The overall findings confirmed that the carrier prevalence among low-risk women in Taiwan is significantly lower than that reported in western countries. Therefore, the most important step for preventing FXS in Taiwan would be to focus on high-risk women by promoting general awareness of this disease and spreading knowledge regarding the benefits of carrier screening and prenatal testing. © 2017 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  5. Analysis of differentially expressed genes in two immunologically distinct strains of Eimeria maxima using suppression subtractive hybridization and dot-blot hybridization

    Science.gov (United States)

    2014-01-01

    Background It is well known that different Eimeria maxima strains exhibit significant antigenic variation. However, the genetic basis of these phenotypes remains unclear. Methods Total RNA and mRNA were isolated from unsporulated oocysts of E. maxima strains SH and NT, which were found to have significant differences in immunogenicity in our previous research. Two subtractive cDNA libraries were constructed using suppression subtractive hybridization (SSH) and specific genes were further analyzed by dot-blot hybridization and qRT-PCR analysis. Results A total of 561 clones were selected from both cDNA libraries and the length of the inserted fragments was 0.25–1.0 kb. Dot-blot hybridization revealed a total of 86 differentially expressed clones (63 from strain SH and 23 from strain NT). Nucleotide sequencing analysis of these clones revealed ten specific contigs (six from strain SH and four from strain NT). Further analysis found that six contigs from strain SH and three from strain NT shared significant identities with previously reported proteins, and one contig was presumed to be novel. The specific differentially expressed genes were finally verified by RT-PCR and qRT-PCR analyses. Conclusions The data presented here suggest that specific genes identified between the two strains may be important molecules in the immunogenicity of E. maxima that may present potential new drug targets or vaccine candidates for coccidiosis. PMID:24894832

  6. Monitoring larval populations of the Douglas-fir tussock moth and the western spruce budworm on permanent plots: sampling methods and statistical properties of data

    Science.gov (United States)

    A.R. Mason; H.G. Paul

    1994-01-01

    Procedures for monitoring larval populations of the Douglas-fir tussock moth and the western spruce budworm are recommended based on many years experience in sampling these species in eastern Oregon and Washington. It is shown that statistically reliable estimates of larval density can be made for a population by sampling host trees in a series of permanent plots in a...

  7. Radiation dose determines the method for quantification of DNA double strand breaks

    International Nuclear Information System (INIS)

    Bulat, Tanja; Keta, Olitija; Korićanac, Lela; Žakula, Jelena; Petrović, Ivan; Ristić-Fira, Aleksandra; Todorović, Danijela

    2016-01-01

    Ionizing radiation induces DNA double strand breaks (DSBs) that trigger phosphorylation of the histone protein H2AX (γH2AX). Immunofluorescent staining visualizes formation of γH2AX foci, allowing their quantification. This method, as opposed to Western blot assay and Flow cytometry, provides more accurate analysis, by showing exact position and intensity of fluorescent signal in each single cell. In practice there are problems in quantification of γH2AX. This paper is based on two issues: the determination of which technique should be applied concerning the radiation dose, and how to analyze fluorescent microscopy images obtained by different microscopes. HTB140 melanoma cells were exposed to γ-rays, in the dose range from 1 to 16 Gy. Radiation effects on the DNA level were analyzed at different time intervals after irradiation by Western blot analysis and immunofluorescence microscopy. Immunochemically stained cells were visualized with two types of microscopes: AxioVision (Zeiss, Germany) microscope, comprising an ApoTome software, and AxioImagerA1 microscope (Zeiss, Germany). Obtained results show that the level of γH2AX is time and dose dependent. Immunofluorescence microscopy provided better detection of DSBs for lower irradiation doses, while Western blot analysis was more reliable for higher irradiation doses. AxioVision microscope containing ApoTome software was more suitable for the detection of γH2AX foci. (author)

  8. Radiation dose determines the method for quantification of DNA double strand breaks

    Energy Technology Data Exchange (ETDEWEB)

    Bulat, Tanja; Keta, Olitija; Korićanac, Lela; Žakula, Jelena; Petrović, Ivan; Ristić-Fira, Aleksandra [University of Belgrade, Vinča Institute of Nuclear Sciences, Belgrade (Serbia); Todorović, Danijela, E-mail: dtodorovic@medf.kg.ac.rs [University of Kragujevac, Faculty of Medical Sciences, Kragujevac (Serbia)

    2016-03-15

    Ionizing radiation induces DNA double strand breaks (DSBs) that trigger phosphorylation of the histone protein H2AX (γH2AX). Immunofluorescent staining visualizes formation of γH2AX foci, allowing their quantification. This method, as opposed to Western blot assay and Flow cytometry, provides more accurate analysis, by showing exact position and intensity of fluorescent signal in each single cell. In practice there are problems in quantification of γH2AX. This paper is based on two issues: the determination of which technique should be applied concerning the radiation dose, and how to analyze fluorescent microscopy images obtained by different microscopes. HTB140 melanoma cells were exposed to γ-rays, in the dose range from 1 to 16 Gy. Radiation effects on the DNA level were analyzed at different time intervals after irradiation by Western blot analysis and immunofluorescence microscopy. Immunochemically stained cells were visualized with two types of microscopes: AxioVision (Zeiss, Germany) microscope, comprising an ApoTome software, and AxioImagerA1 microscope (Zeiss, Germany). Obtained results show that the level of γH2AX is time and dose dependent. Immunofluorescence microscopy provided better detection of DSBs for lower irradiation doses, while Western blot analysis was more reliable for higher irradiation doses. AxioVision microscope containing ApoTome software was more suitable for the detection of γH2AX foci. (author)

  9. Detection of mutations related to drug resistance in M. tuberculosis by dot blot hybridization and spoligotyping using specific radiolabelled probes

    International Nuclear Information System (INIS)

    El-Maghraby, T.K.; Abdelazeim, O.

    2002-01-01

    The present work has been conducted to determine the mutations related to drug resistance in M. tuberculosis in 63 Egyptian isolates using dot blot hybridization and spoligotyping. The PCR was done for amplification rpoB and katG genes in isolates. Dot blot hybridization were done to PCR products by using specific radiolabelled probes. Moreover, spoligotyping was done to know about the different strains found in Egypt. The results revealed that 58% from isolates had drug resistance to one or more of antituberculosis drugs. The results of spoligotyping have revealed that some Egyptian isolates are identical with the international code while the rest has not been identified yet. DNA sequencing was done to identify the mutation that not clear in dot blot hybridization. Early diagnosis of geno typing resistance to antituberculosis drugs is important as well as allow appropriate early patients management with few days of TB diagnosis. Using such strategy for early diagnosis of TB drug resistance allow and fast and potent patient's management

  10. Determinants of Primary School Non-Enrollment and Absenteeism: Results from a Retrospective, Convergent Mixed Methods, Cohort Study in Rural Western Kenya.

    Directory of Open Access Journals (Sweden)

    Nia King

    Full Text Available Education is a key element in the socioeconomic development required to improve quality of life in Kenya. Despite the introduction of free primary education, primary school enrollment and attendance levels remain low. Drawing on qualitative and quantitative data, this study explores the determinants of non-enrollment and absenteeism in rural western Kenya and potential mitigation strategies to address these issues.The study was conducted in Bwaliro village in rural western Kenya. A random sample of 64 students was obtained by blocking the village primary school's student population according to grade level, gender, and orphan status. Qualitative and quantitative data were collected through interviews with parents, guardians, and key informants, and focus group discussions with students. Quantitative data were compared using chi-square tests, Student's T-test, and Poisson regressions. Qualitative data were analyzed using thematic content analysis.Malaria, menstruation, and lack of money were among the most notable determinants of primary school dropout and absenteeism, and these factors disproportionately impacted orphans and female students. Potential mitigation strategies suggested by the community included provision of malaria treatment or prevention, reduction in education costs, expansion of the established school-feeding program, and provision of sanitary pads.Despite free primary education, numerous factors continue to prevent children in rural western Kenya from attending primary school. The findings suggest that interventions should primarily target orphaned and female students. Prior to implementation, suggested mitigation strategies should be assessed for cost-effectiveness.

  11. Sulphate reduction and vertical distribution of sulphate-reducing bacteria quantified by rRNA slot-blot hybridization in a coastal marine sediment

    DEFF Research Database (Denmark)

    Sahm, K.; MacGregor, BJ; Jørgensen, BB

    1999-01-01

    In the past, enumeration of sulphate-reducing bacteria (SRB) by cultivation-based methods generally contradicted measurements of sulphate reduction, suggesting unrealistically high respiration rates per cell. Here, we report evidence that quantification of SRB rRNA by slot-blot hybridization...... between 18% and 25% to the prokaryotic rRNA pool. The dominant SRB were related to complete oxidizing genera (Desulphococcus, Desulphosarcina and Desulphobacterium), while Desulpho-bacter could not be detected. The vertical profile and quantity of rRNA from SRB was compared with sulphate reduction rates......, directly above the sulphate reduction maximum. Cell numbers calculated by converting the relative contribution of SRB rRNA to the percentage of DAPI-stained cells indicated a population size for SRB of 2.4-6.1 x 10(8) cells cm(-3) wet sediment. Cellular sulphate reduction rates calculated on the basis...

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

    Energy Technology Data Exchange (ETDEWEB)

    Aravalli, Rajagopal N., E-mail: aravalli@umn.edu [Department of Radiology, University of Minnesota Medical School, MMC 292, 420 Delaware Street SE, Minneapolis, MN 55455 (United States); Park, Chang W. [Department of Medicine, University of Minnesota Medical School, MMC 36, 420 Delaware Street SE, Minneapolis, MN 55455 (United States); Steer, Clifford J., E-mail: steer001@umn.edu [Department of Medicine, University of Minnesota Medical School, MMC 36, 420 Delaware Street SE, Minneapolis, MN 55455 (United States); Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN 55455 (United States)

    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.

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

    International Nuclear Information System (INIS)

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

    2016-01-01

    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.

  14. Mere end blot en bid af hverdagen- Måltidet i et leve- og bomiljø

    DEFF Research Database (Denmark)

    Bundgaard, Karen Marie

    2005-01-01

    . Datamaterialet bygger på deltagerobservationer og interviews. Undersøgelsen viste, at den måde måltiderne var organiseret på gav tid og rum til en hjemlig atmosfære, til et levende fællesskab, til det at være noget og at være sig selv og til at have værdifulde gøremål. Måltiderne var ikke blot en bid – men en...

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

    Science.gov (United States)

    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

  16. Interpretation of biological and mechanical variations between the Lowry versus Bradford method for protein quantification.

    Science.gov (United States)

    Lu, Tzong-Shi; Yiao, Szu-Yu; Lim, Kenneth; Jensen, Roderick V; Hsiao, Li-Li

    2010-07-01

    The identification of differences in protein expression resulting from methodical variations is an essential component to the interpretation of true, biologically significant results. We used the Lowry and Bradford methods- two most commonly used methods for protein quantification, to assess whether differential protein expressions are a result of true biological or methodical variations. MATERIAL #ENTITYSTARTX00026; Differential protein expression patterns was assessed by western blot following protein quantification by the Lowry and Bradford methods. We have observed significant variations in protein concentrations following assessment with the Lowry versus Bradford methods, using identical samples. Greater variations in protein concentration readings were observed over time and in samples with higher concentrations, with the Bradford method. Identical samples quantified using both methods yielded significantly different expression patterns on Western blot. We show for the first time that methodical variations observed in these protein assay techniques, can potentially translate into differential protein expression patterns, that can be falsely taken to be biologically significant. Our study therefore highlights the pivotal need to carefully consider methodical approaches to protein quantification in techniques that report quantitative differences.

  17. Prediction of the optimum hybridization conditions of dot-blot-SNP analysis using estimated melting temperature of oligonucleotide probes.

    Science.gov (United States)

    Shiokai, Sachiko; Kitashiba, Hiroyasu; Nishio, Takeshi

    2010-08-01

    Although the dot-blot-SNP technique is a simple cost-saving technique suitable for genotyping of many plant individuals, optimization of hybridization and washing conditions for each SNP marker requires much time and labor. For prediction of the optimum hybridization conditions for each probe, we compared T (m) values estimated from nucleotide sequences using the DINAMelt web server, measured T (m) values, and hybridization conditions yielding allele-specific signals. The estimated T (m) values were comparable to the measured T (m) values with small differences of less than 3 degrees C for most of the probes. There were differences of approximately 14 degrees C between the specific signal detection conditions and estimated T (m) values. Change of one level of SSC concentrations of 0.1, 0.2, 0.5, and 1.0x SSC corresponded to a difference of approximately 5 degrees C in optimum signal detection temperature. Increasing the sensitivity of signal detection by shortening the exposure time to X-ray film changed the optimum hybridization condition for specific signal detection. Addition of competitive oligonucleotides to the hybridization mixture increased the suitable hybridization conditions by 1.8. Based on these results, optimum hybridization conditions for newly produced dot-blot-SNP markers will become predictable.

  18. Label-free DNA quantification via a 'pipette, aggregate and blot' (PAB) approach with magnetic silica particles on filter paper.

    Science.gov (United States)

    Li, Jingyi; Liu, Qian; Alsamarri, Hussein; Lounsbury, Jenny A; Haversitick, Doris M; Landers, James P

    2013-03-07

    Reliable measurement of DNA concentration is essential for a broad range of applications in biology and molecular biology, and for many of these, quantifying the nucleic acid content is inextricably linked to obtaining optimal results. In its most simplistic form, quantitative analysis of nucleic acids can be accomplished by UV-Vis absorbance and, in more sophisticated format, by fluorimetry. A recently reported new concept, the 'pinwheel assay', involves a label-free approach for quantifying DNA through aggregation of paramagnetic beads in a rotating magnetic field. Here, we describe a simplified version of that assay adapted for execution using only a pipet and filter paper. The 'pipette, aggregate, and blot' (PAB) approach allows DNA to induce bead aggregation in a pipette tip through exposure to a magnetic field, followed by dispensing (blotting) onto filter paper. The filter paper immortalises the extent of aggregation, and digital images of the immortalized bead conformation, acquired with either a document scanner or a cell phone camera, allows for DNA quantification using a noncomplex algorithm. Human genomic DNA samples extracted from blood are quantified with the PAB approach and the results utilized to define the volume of sample used in a PCR reaction that is sensitive to input mass of template DNA. Integrating the PAB assay with paper-based DNA extraction and detection modalities has the potential to yield 'DNA quant-on-paper' devices that may be useful for point-of-care testing.

  19. Mere end blot pirringer

    DEFF Research Database (Denmark)

    Jantzen, Christian; Østergaard, Per

    2006-01-01

    Med udgangspunkt i en række cases på vellykkede oplevelsesøkonomiske forretningsmodeller argumenterer artiklen for, at oplevelsesprodukter skal bygge på et klart tema, som forbrugeren kan koble sig sanse- og følelsesmæssigt op på. Forbrugeren skal kunne omsætte produktets pirringer til egne erfar...

  20. Ej blot til lyst

    DEFF Research Database (Denmark)

    Leroyer, Patrick

    2010-01-01

    The purpose of this article is to reassert the crucial importance of access to data in lexicographic information tools and, expanding on this, to establish the existence of two distinct lexicographic access modes - consultation and navigation. It is explained how the tools can be decalibrated whe...... balance between user, access, and data is disturbed, and how access to data then is jeopardized. Taking online wine guides as a case in point, it is shown how such multifunctional information tools do benefit from a lexicographic design featuring both access modes....

  1. Evaluation of a new eastern blotting technique for the analysis of ginsenoside Re in American ginseng berry pulp extracts.

    Science.gov (United States)

    Morinaga, Osamu; Uto, Takuhiro; Yuan, Chun-Su; Tanaka, Hiroyuki; Shoyama, Yukihiro

    2010-06-01

    A new eastern blotting technique has been established for ginsenoside Re (G-Re) contained in American ginseng berry pulp extracts. G-Re in American ginseng berry pulp was extracted using 100% methanol, 100% ethanol, 50% aqueous methanol, and 50% aqueous ethanol. The combined crude extracts were applied onto a polyethersulfone membrane and developed using the methanol-water-acetic acid solvent system (45:55:1 v/v). Separated components were immunostained using anti-G-Re monoclonal antibody. G-Re was first specifically detected and then quantitatively analyzed using NIH Imaging software. We also confirmed that the most suitable solvent was 50% aqueous methanol for extracting G-Re from American ginseng berry pulp. (c) 2009 Elsevier B.V. All rights reserved.

  2. Modifying a standard method allows simultaneous extraction of RNA and protein, enabling detection of enzymes in the rat retina with low expressions and protein levels.

    Science.gov (United States)

    Agardh, Elisabet; Gustavsson, Carin; Hagert, Per; Nilsson, Marie; Agardh, Carl-David

    2006-02-01

    The aim of the study was to evaluate messenger RNA and protein expression in limited amounts of tissue with low protein content. The Chomczynski method was used for simultaneous extraction of RNA, and protein was modified in the protein isolation step. Template mass and cycling time for the complementary DNA synthesis step of real-time reverse transcription-polymerase chain reaction (RT-PCR) for analysis of catalase, copper/zinc superoxide dismutase, manganese superoxide dismutase, the catalytic subunit of glutamylcysteine ligase, glutathione peroxidase 1, and the endogenous control cyclophilin B (CypB) were optimized before PCR. Polymerase chain reaction accuracy and efficacy were demonstrated by calculating the regression (R2) values of the separate amplification curves. Appropriate antibodies, blocking buffers, and running conditions were established for Western blot, and protein detection and multiplex assays with CypB were performed for each target. During the extraction procedure, the protein phase was dissolved in a modified washing buffer containing 0.1% sodium dodecyl sulfate, followed by ultrafiltration. Enzyme expression on real-time RT-PCR was accomplished with high reliability and reproducibility (R2, 0.990-0.999), and all enzymes except for glutathione peroxidase 1 were detectable in individual retinas on Western blot. Western blot multiplexing with CypB was possible for all targets. In conclusion, connecting gene expression directly to protein levels in the individual rat retina was possible by simultaneous extraction of RNA and protein. Real-time RT-PCR and Western blot allowed accurate detection of retinal protein expressions and levels.

  3. Southern blot analysis of skin biopsies for human papillomavirus DNA: renal allograft recipients in south-eastern Queensland.

    Science.gov (United States)

    Trenfield, K; Salmond, C A; Pope, J H; Hardie, I R

    1993-01-01

    The 104 skin biopsies from 34 patients who attended a Renal Transplant Unit in Brisbane over 12 months included 40 squamous cell carcinoma (SCC), 22 solar keratoses, 4 hyperkeratoses, 18 warts and 11 basal cell carcinoma (BCC). Human papillomavirus (HPV) DNA was identified by Southern blot hybridisation using, as individual probes, purified insert DNA from recombinant HPV 1, 2, 3 or 3/10, 4, 5 or 5/8, 7, 11, 16, 18 and 41 under relaxed conditions and characterised by restriction enzyme analysis and Southern blot hybridisation under more stringent conditions. Genomic HPV DNA was characterised in 7 skin biopsies from 4 renal allograft recipients (RARs): HPV 1A in a SCC (20 copies/cell) and a BCC (10 copies/cell) from the one patient, HPV 36 (20 copies/cell) in a SCC, HPV 1A [symbol: see text] 1000 copies/cell) in a wart and HPV 2B (200-800 copies/cell) in 3 warts from the one patient. Only HPV 1A in the SCC exhibited a significant degree of subtype variation. HPV DNA was identified in another 5 skin biopsies from another 4 RARs: HPV 3A in a wart and a hyperkeratosis, HPV 3/10-related DNA in 2 solar keratoses and HPV 5/8-related DNA in another (20-50 copies/cell). The incidence of HPV 5 (or 5-related HPVs) in RAR SCC was very low and that of HPV DNA in RAR warts was lower than that recorded elsewhere but this was not due to insensitivity of the assays. There was no evidence for a role for HPV in the aetiology of skin cancer in RARs in south-eastern Queensland but the possibility remains that as yet unidentified HPV types are involved.

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

    International Nuclear Information System (INIS)

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

    1988-01-01

    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 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

  5. Method of Relating Grain Size Distribution to Hydraulic Conductivity in Dune Sands to Assist in Assessing Managed Aquifer Recharge Projects: Wadi Khulays Dune Field, Western Saudi Arabia

    Directory of Open Access Journals (Sweden)

    Oliver M. Lopez

    2015-11-01

    Full Text Available Planning for use of a dune field aquifer for managed aquifer recharge (MAR requires that hydraulic properties need to be estimated over a large geographic area. Saturated hydraulic conductivity of dune sands is commonly estimated from grain size distribution data by employing some type of empirical equation. Over 50 samples from the Wadi Khulays dune field in Western Saudi Arabia were collected and the grain size distribution, porosity, and hydraulic conductivity were measured. An evaluation of 20 existing empirical equations showed a generally high degree of error in the predicted compared to the measured hydraulic conductivity values of these samples. Statistical analyses comparing estimated versus measured hydraulic conductivity demonstrated that there is a significant relationship between hydraulic conductivity and mud percentage (and skewness. The modified Beyer equation, which showed a generally low prediction error, was modified by adding a second term fitting parameter related to the mud concentration based on 25 of the 50 samples analyzed. An inverse optimization process was conducted to quantify the fitting parameter and a new empirical equation was developed. This equation was tested against the remaining 25 samples analyzed and produced an estimated saturated hydraulic conductivity with the lowest error of any empirical equation. This methodology can be used for large dune field hydraulic conductivity estimation and reduce planning costs for MAR systems.

  6. Method of Relating Grain Size Distribution to Hydraulic Conductivity in Dune Sands to Assist in Assessing Managed Aquifer Recharge Projects: Wadi Khulays Dune Field, Western Saudi Arabia

    KAUST Repository

    Lopez Valencia, Oliver Miguel

    2015-11-12

    Planning for use of a dune field aquifer for managed aquifer recharge (MAR) requires that hydraulic properties need to be estimated over a large geographic area. Saturated hydraulic conductivity of dune sands is commonly estimated from grain size distribution data by employing some type of empirical equation. Over 50 samples from the Wadi Khulays dune field in Western Saudi Arabia were collected and the grain size distribution, porosity, and hydraulic conductivity were measured. An evaluation of 20 existing empirical equations showed a generally high degree of error in the predicted compared to the measured hydraulic conductivity values of these samples. Statistical analyses comparing estimated versus measured hydraulic conductivity demonstrated that there is a significant relationship between hydraulic conductivity and mud percentage (and skewness). The modified Beyer equation, which showed a generally low prediction error, was modified by adding a second term fitting parameter related to the mud concentration based on 25 of the 50 samples analyzed. An inverse optimization process was conducted to quantify the fitting parameter and a new empirical equation was developed. This equation was tested against the remaining 25 samples analyzed and produced an estimated saturated hydraulic conductivity with the lowest error of any empirical equation. This methodology can be used for large dune field hydraulic conductivity estimation and reduce planning costs for MAR systems.

  7. A comparison of DNA extraction protocols from blood spotted on FTA cards for the detection of tick-borne pathogens by Reverse Line Blot hybridization.

    Science.gov (United States)

    Hailemariam, Zerihun; Ahmed, Jabbar Sabir; Clausen, Peter-Henning; Nijhof, Ard Menzo

    2017-01-01

    An essential step in the molecular detection of tick-borne pathogens (TBPs) in blood is the extraction of DNA. When cooled storage of blood under field conditions prior to DNA extraction in a dedicated laboratory is not possible, the storage of blood on filter paper forms a promising alternative. We evaluated six DNA extraction methods from blood spotted on FTA Classic ® cards (FTA cards), to determine the optimal protocol for the subsequent molecular detection of TBPs by PCR and the Reverse Line Blot hybridization assay (RLB). Ten-fold serial dilutions of bovine blood infected with Babesia bovis, Theileria mutans, Anaplasma marginale or Ehrlichia ruminantium were made by dilution with uninfected blood and spotted on FTA cards. Subsequently, DNA was extracted from FTA cards using six different DNA extraction protocols. DNA was also isolated from whole blood dilutions using a commercial kit. PCR/RLB results showed that washing of 3mm discs punched from FTA cards with FTA purification reagent followed by DNA extraction using Chelex ® resin was the most sensitive procedure. The detection limit could be improved when more discs were used as starting material for the DNA extraction, whereby the use of sixteen 3mm discs proved to be most practical. The presented best practice method for the extraction of DNA from blood spotted on FTA cards will facilitate epidemiological studies on TBPs. It may be particularly useful for field studies where a cold chain is absent. Copyright © 2016 Elsevier GmbH. All rights reserved.

  8. Simultaneous and independent detection of C9ORF72 alleles with low and high number of GGGGCC repeats using an optimised protocol of Southern blot hybridisation.

    Science.gov (United States)

    Buchman, Vladimir L; Cooper-Knock, Johnathan; Connor-Robson, Natalie; Higginbottom, Adrian; Kirby, Janine; Razinskaya, Olga D; Ninkina, Natalia; Shaw, Pamela J

    2013-04-08

    Sizing of GGGGCC hexanucleotide repeat expansions within the C9ORF72 locus, which account for approximately 10% of all amyotrophic lateral sclerosis (ALS) cases, is urgently required to answer fundamental questions about mechanisms of pathogenesis in this important genetic variant. Currently employed PCR protocols are limited to discrimination between the presence and absence of a modified allele with more than 30 copies of the repeat, while Southern hybridisation-based methods are confounded by the somatic heterogeneity commonly present in blood samples, which might cause false-negative or ambiguous results. We describe an optimised Southern hybridisation-based protocol that allows confident detection of the presence of a C9ORF72 repeat expansion alongside independent assessment of its heterogeneity and the number of repeat units. The protocol can be used with either a radiolabeled or non-radiolabeled probe. Using this method we have successfully sized the C9ORF72 repeat expansion in lymphoblastoid cells, peripheral blood, and post-mortem central nervous system (CNS) tissue from ALS patients. It was also possible to confidently demonstrate the presence of repeat expansion, although of different magnitude, in both C9ORF72 alleles of the genome of one patient. The suggested protocol has sufficient advantages to warrant adoption as a standard for Southern blot hybridisation analysis of GGGGCC repeat expansions in the C9ORF72 locus.

  9. Cross-Sectional Serological Survey of Human Fascioliasis in Canutama Municipality in Western Amazon, Brazil

    Directory of Open Access Journals (Sweden)

    Marcel Gonçalves Maciel

    2018-01-01

    Full Text Available Background. Fascioliasis is an important parasitic disease. In the northern region of Brazil, a human parasite infection has been reported through a coprological survey. Eggs of Fasciola hepatica were found in fecal samples of 11 individuals. Knowledge of the infection in animals or the presence of snails is necessary to address the possibility of the parasite cycle occurrence in that region. The aim of this study was to describe the transmission of human fascioliasis in Canutama, Amazonas, in Western Amazonia, Brazil. Methods. Serological (ELISA and Western Blot, WB and parasitological analyses were carried out in humans. In addition, the presence of the intermediate snail host within the community was examined. Results. A total of 434 human samples were included in the study, of which 36 (8.3% were reactive by ELISA and 8 (1.8% were reactive by WB. Fasciola hepatica eggs were found in one human sample. The occurrence of the intermediated host was recorded and 31/43 specimens were identified as Lymnaea columella. Conclusion. Canutama constitutes a focus of transmission of human fascioliasis. This study describes the first serological survey for human fascioliasis, as well as its simultaneous occurrence in human hosts and possible intermediates performed in northern Brazil.

  10. Teaching the Western.

    Science.gov (United States)

    Lenihan, John H.

    2002-01-01

    Discusses the content of a course on the genre of western films that was utilized as a film study and a U.S. cultural history credit. Describes in detail the film, "Winchester '73," and addresses other films utilized in the course. States that the course also focuses on the development of the western genre. (CMK)

  11. Western Slope Colorado

    International Nuclear Information System (INIS)

    Epis, R.C.; Callender, J.F.

    1981-01-01

    A conference on the geology and geologic resources of the Western Slope of western Colorado and eastern Utah is presented. Fourteen papers from the conference have been abstracted and indexed for the Department of Energy's Energy Data Base. These papers covered such topics as uranium resources, oil shale deposits, coal resources, oil and gas resources, and geothermal resources of the area

  12. "They just scraped off the calluses": a mixed methods exploration of foot care access and provision for people with rheumatoid arthritis in south-western Sydney, Australia.

    Science.gov (United States)

    Hendry, Gordon J; Gibson, Kathryn A; Pile, Kevin; Taylor, Luke; Du Toit, Verona; Burns, Joshua; Rome, Keith

    2013-08-13

    There is little indication that foot health services in Australia are meeting modern day recommendations for Rheumatoid Arthritis (RA) patients. The overall objective of this study was to explore the current state of foot health services for patients with RA with an emphasis on identifying barriers to the receipt of appropriate foot care in South-West Sydney, New South Wales, Australia. A mixed (quantitative and qualitative) approach was adopted. Indications for appropriate access to foot care were determined by comparing the foot health, disease and socio-demographic characteristics of patients with unmet foot care demands, foot care users and patients with no demands for foot care. Perceptions of provision of, and access to, foot care were explored by conducting telephone-based interviews using an interpretative phenomenology approach with thematic analysis. Twenty-nine participants took part in the cross-sectional quantitative research study design, and 12 participants took part in the interpretative phenomenological approach (qualitative study). Foot care access appeared to be driven predominantly by the presence of rearfoot deformity, which was significantly worse amongst participants in the foot care user group (p = 0.02). Five main themes emerged from the qualitative data: 1) impact of disease-related foot symptoms, 2) footwear difficulties, 3) medical/rheumatology encounters, 4) foot and podiatry care access and experiences, and 5) financial hardship. Foot care provision does not appear to be driven by appropriate foot health characteristics such as foot pain or foot-related disability. There may be significant shortfalls in footwear and foot care access and provision in Greater Western Sydney. Several barriers to adequate foot care access and provision were identified and further efforts are required to improve access to and the quality of foot care for people who have RA. Integration of podiatry services within rheumatology centres could resolve unmet

  13. Evaluating a Local Ensemble Transform Kalman Filter snow cover data assimilation method to estimate SWE within a high-resolution hydrologic modeling framework across Western US mountainous regions

    Science.gov (United States)

    Oaida, C. M.; Andreadis, K.; Reager, J. T., II; Famiglietti, J. S.; Levoe, S.

    2017-12-01

    Accurately estimating how much snow water equivalent (SWE) is stored in mountainous regions characterized by complex terrain and snowmelt-driven hydrologic cycles is not only greatly desirable, but also a big challenge. Mountain snowpack exhibits high spatial variability across a broad range of spatial and temporal scales due to a multitude of physical and climatic factors, making it difficult to observe or estimate in its entirety. Combing remotely sensed data and high resolution hydrologic modeling through data assimilation (DA) has the potential to provide a spatially and temporally continuous SWE dataset at horizontal scales that capture sub-grid snow spatial variability and are also relevant to stakeholders such as water resource managers. Here, we present the evaluation of a new snow DA approach that uses a Local Ensemble Transform Kalman Filter (LETKF) in tandem with the Variable Infiltration Capacity macro-scale hydrologic model across the Western United States, at a daily temporal resolution, and a horizontal resolution of 1.75 km x 1.75 km. The LETKF is chosen for its relative simplicity, ease of implementation, and computational efficiency and scalability. The modeling/DA system assimilates daily MODIS Snow Covered Area and Grain Size (MODSCAG) fractional snow cover over, and has been developed to efficiently calculate SWE estimates over extended periods of time and covering large regional-scale areas at relatively high spatial resolution, ultimately producing a snow reanalysis-type dataset. Here we focus on the assessment of SWE produced by the DA scheme over several basins in California's Sierra Nevada Mountain range where Airborne Snow Observatory data is available, during the last five water years (2013-2017), which include both one of the driest and one of the wettest years. Comparison against such a spatially distributed SWE observational product provides a greater understanding of the model's ability to estimate SWE and SWE spatial variability

  14. Comparison of brown sugar, hot water, and salt methods for detecting western cherry fruit fly (Diptera: Tephritidae) larvae in sweet cherry

    Science.gov (United States)

    Brown sugar or hot water methods have been developed to detect larvae of tephritid fruit flies in post-harvest fruit in order to maintain quarantine security. It would be useful to determine if variations of these methods can yield better results and if less expensive alternatives exist. This stud...

  15. Dot Blot para determinar la identidad antigénica en vacunas conjugadas contra Streptococcus pneumoniae serotipo 19F

    Directory of Open Access Journals (Sweden)

    Osmir Cabrera-Blanco

    2017-04-01

    Full Text Available Las autoridades regulatorias recomiendan el uso de técnicas de Resonancia Magnética Nuclear o técnicas serológicas para la determinación de la identidad de los antígenos presentes en las vacunas conjugadas. Con la aparición de las vacunas conjugadas multivalentes, se ha hecho necesario recurrir a técnicas inmunoquímicas con la utilización de anticuerpos monoclonales para aumentar la sensibilidad en la determinación de la identidad de los antígenos en dichas vacunas conjugadas. El objetivo del presente trabajo fue establecer las condiciones óptimas de trabajo que permitieran utilizar la técnica del Dot Blot para determinar la identidad de los antígenos en vacunas conjugadas de Streptococcus pneumoniae serotipo 19F. Para ello se estudiaron los tiempos de incubación, la influencia del reactivo en la solución de bloqueo; también las concentraciones óptimas del anticuerpo monoclonal y de los ingredientes farmacéuticos activos, así como los volúmenes de aplicación óptimos para estos y vacunas. Se utilizó un anticuerpo monoclonal contra el polisacárido capsular del serotipo 19F de neumococo. Las muestras empleadas en este trabajo fueron lotes de ingredientes farmacéuticos activos de conjugados de polisacárido capsular 19F y lotes de un candidato vacunal cubano conjugado heptavalente contra neumococos. Los resultados mostraron que para la determinación de la identidad antigénica fueron suficientes 10 µL de muestras de los principios activos a una concentración de 125 µg/mL e igual volumen para las vacunas heptavalentes. Quedó demostrado que una concentración de 1 µg/mL para el anticuerpo monoclonal y tiempos de incubación de 30 min a 37 °C fueron suficientes para la determinación. Estos resultados permiten concluir que quedaron establecidas las condiciones óptimas de trabajo para determinar la identidad antigénica por Dot Blot del polisacárido capsular de S. pneumoniae serotipo 19F presente en las vacunas

  16. Soil classification for seismic site effect using MASW and ReMi methods: A case study from western Anatolia (Dikili -İzmir)

    Science.gov (United States)

    Karabulut, Savaş

    2018-03-01

    The study area is located in the northern part of Izmir, Western Turkey, prone to an active tectonic extensional regime and includes typical features of sedimentary basins, horst-grabens surrounded by a series of normal and strike-slip faults. In September 1939 the Dikili (Kabakum) earthquake with a magnitude of Mw: 6.6 occurred and after this phenomenon, residents moved from the west of Dikili to the east (i.d. soft sediments to relative to rock area). A proper estimate of the earthquake-related hazard for the area is the main objective of this study. The site effect and soil engineering problems for estimating hazard parameters at the soil surface need to be carefully analyzed for seismic site classification and geo-engineering problems like soil liquefaction, soil settlement, soil bearing capacity and soil amplification. To solve the soil static and dynamic problems, shear-wave velocities have been used in a joint interpretation process; Multichannel Analysis of Surface Waves (MASW) and Refraction Microtremor (ReMi) analyses were conducted on 121 sites with 300 × 300 m grid size in an area of 60 km2. It has been proposed that the probability of an earthquake with a magnitude of Mw: 6 occurring within 10 years is 64%, when considering the Gutenberg-Richter model. This puts the region under an important earthquake risk. The estimated Vs30 values are ≤180 m/s in the central and the northernmost part of the study area are showing an E type soil after the classification of NEHRP, where alluvial deposits are dominant. Vs30 values in the north and central part are between 180 ≤ Vs ≤ 360 m/s suggesting a D type soil. In the southernmost part of the study area where volcanic rocks are widely distributed, Vs30 values range between 360 and 908 m/s, corresponding to a C type and B type soil. The results show that soil liquefaction induced settlement and soil amplification are the most important problems in the south and the northernmost part of the study area, which

  17. Factors affecting the use of prenatal care by non-western women in industrialized western countries: a systematic review.

    OpenAIRE

    Boerleider, A.W.; Wiegers, T.A.; Manniën, J.; Francke, A.L.; Devillé, W.L.J.M.

    2013-01-01

    Background: Despite the potential of prenatal care for addressing many pregnancy complications and concurrent health problems, non-western women in industrialized western countries more often make inadequate use of prenatal care than women from the majority population do. This study aimed to give a systematic review of factors affecting non-western women's use of prenatal care (both medical care and prenatal classes) in industrialized western countries.Methods: Eleven databases (PubMed, Embas...

  18. Application of the Reverse Line Blot Assay for the Molecular Detection of Theileria and Babesia sp. in Sheep and Goat Blood Samples from Pakistan

    Directory of Open Access Journals (Sweden)

    A Rasul

    2013-06-01

    Full Text Available Background: The present study was designed to detect the presence of tick-borne parasites (Theileria and Babesia spp. in 196 blood samples collected from apparently healthy sheep and goats from two provinces, Punjab and Khyber Pukhtoon Khwa, in Pakistan.Methods: Reverse line blot (RLB assay was applied for the parasitic detection by the amplification of hypervariable V4 region of the 18S ribosomal RNA (rRNA gene. A membrane with covalently linked generic and species specific oligonucleotide probes was used for the hybridization of amplified PCR products.Results: Parasites were detected in 16% of the ruminant blood samples under study. Two Theileria species, T. lestoquardi and T. ovis, were identified in samples. 25, of the total 32, infected animals were from Khyber Pukhtoon Khwa.Conclusion: Sheep were more prone to tick borne haemoprotozans as 81% infected samples were sheep as compared to 19% goats (P > 0.001. Risk factor analysis revealed that male (P = 0.03, ani­mals infested by ticks (P = 0.03 and herd composed of sheep only (P = 0.001 were more infected by blood parasites.

  19. Violence the Western way.

    Science.gov (United States)

    Roth, B E

    1997-10-01

    Despite the quiet revolution in response to changing conceptualizations of gender in psychoanalysis, the Western has remained the domain of aggressive phallic masculinity. The iconic imagery of the Western, when combined with its narrative trajectory, is used to tell stories of violent encounters between men. The acceptance of the genre, and its duplication by other cultures and film makers, indicates that the Westerns' imagery and moral solutions tap into some basic deep structures of anxiety and pleasure in violence between men. As long as societies require subtle sublimations of aggressive and violent drives, it is likely that men will seek imaginary regressive experiences to discharge frustrations.

  20. Comparison of first order analysis and Monte Carlo methods in evaluating groundwater model uncertainty: a case study from an iron ore mine in the Pilbara Region of Western Australia

    Science.gov (United States)

    Firmani, G.; Matta, J.

    2012-04-01

    The expansion of mining in the Pilbara region of Western Australia is resulting in the need to develop better water strategies to make below water table resources accessible, manage surplus water and deal with water demands for processing ore and construction. In all these instances, understanding the local and regional hydrogeology is fundamental to allow sustainable mining; minimising the impacts to the environment. An understanding of the uncertainties of the hydrogeology is necessary to quantify the risks and make objective decisions rather than relying on subjective judgements. The aim of this paper is to review some of the methods proposed by the published literature and find approaches that can be practically implemented in an attempt to estimate model uncertainties. In particular, this paper adopts two general probabilistic approaches that address the parametric uncertainty estimation and its propagation in predictive scenarios: the first order analysis and Monte Carlo simulations. A case example application of the two techniques is also presented for the dewatering strategy of a large below water table open cut iron ore mine in the Pilbara region of Western Australia. This study demonstrates the weakness of the deterministic approach, as the coefficients of variation of some model parameters were greater than 1.0; and suggests a review of the model calibration method and conceptualisation. The uncertainty propagation into predictive scenarios was calculated assuming the parameters with a coefficient of variation higher than 0.25 as deterministic, due to computational difficulties to achieve an accurate result with the Monte Carlo method. The conclusion of this case study was that the first order analysis appears to be a successful and simple tool when the coefficients of variation of calibrated parameters are less than 0.25.

  1. Detection of Zika virus using reverse-transcription LAMP coupled with reverse dot blot analysis in saliva.

    Directory of Open Access Journals (Sweden)

    Maite Sabalza

    Full Text Available In recent years, there have been increasing numbers of infectious disease outbreaks that spread rapidly to population centers resulting from global travel, population vulnerabilities, environmental factors, and ecological disasters such as floods and earthquakes. Some examples of the recent outbreaks are the Ebola epidemic in West Africa, Middle East respiratory syndrome coronavirus (MERS-Co in the Middle East, and the Zika outbreak through the Americas. We have created a generic protocol for detection of pathogen RNA and/or DNA using loop-mediated isothermal amplification (LAMP and reverse dot-blot for detection (RDB and processed automatically in a microfluidic device. In particular, we describe how a microfluidic assay to detect HIV viral RNA was converted to detect Zika virus (ZIKV RNA. We first optimized the RT-LAMP assay to detect ZIKV RNA using a benchtop isothermal amplification device. Then we implemented the assay in a microfluidic device that will allow analyzing 24 samples simultaneously and automatically from sample introduction to detection by RDB technique. Preliminary data using saliva samples spiked with ZIKV showed that our diagnostic system detects ZIKV RNA in saliva. These results will be validated in further experiments with well-characterized ZIKV human specimens of saliva. The described strategy and methodology to convert the HIV diagnostic assay and platform to a ZIKV RNA detection assay provides a model that can be readily utilized for detection of the next emerging or re-emerging infectious disease.

  2. The Western Denmark Cardiac Computed Tomography Registry

    DEFF Research Database (Denmark)

    Nielsen, Lene Hüche; Nørgaard, Bjarne Linde; Tilsted, Hans-Henrik

    2014-01-01

    BACKGROUND: As a subregistry to the Western Denmark Heart Registry (WDHR), the Western Denmark Cardiac Computed Tomography Registry (WDHR-CCTR) is a clinical database established in 2008 to monitor and improve the quality of cardiac computed tomography (CT) in Western Denmark. OBJECTIVE: We...... examined the content, data quality, and research potential of the WDHR-CCTR. METHODS: We retrieved 2008-2012 data to examine the 1) content; 2) completeness of procedure registration using the Danish National Patient Registry as reference; 3) completeness of variable registration comparing observed vs...

  3. Can cognitive dissonance methods developed in the West for combatting the ‘thin ideal’ help slow the rapidly increasing prevalence of eating disorders in non-Western cultures?

    Science.gov (United States)

    Witcomb, Gemma L.; Arcelus, Jon; Chen, Jue

    2013-01-01

    Summary Eating disorders are common, life-threatening conditions in Western countries, but until relatively recently they were regarded as uncommon in non-Western cultures. However, the prevalence of eating disorders in many of the more affluent non-Western countries is rising rapidly as community members, particularly young women, internalize the ‘thin ideal’ that has been widely promoted by the international media. This review discusses the factors involved in the development of eating disorders in non-Western settings with a particular emphasis on the influences of urbanization, modernization, Westernization, and the resulting changes in women's roles. The cognitive dissonance programs developed in Western countries that have proven successful in countering the negative effects of the thin idea are described and their potential application to East Asia and other non-Western countries are discussed. PMID:24991176

  4. Western Mountain Initiative - Background

    Science.gov (United States)

    unprecendented severity in the western United States, extensive tree mortality from outbreaks of bark beetles climatic stressors (Goals 1.1, 1.3) and identification of critical areas (Goal 1.2). Causal mechanisms

  5. Assessment methods as effective tools for learning outcomes of students in senior secondary schools in Ila-Orangun, south western Nigeria

    Directory of Open Access Journals (Sweden)

    Lamidi W.A

    2013-06-01

    Full Text Available Different methods of assessment on the students learning outcomes in Agricultural Science at five different secondary schools in Ila-Orangun, Osun State were studied. An arm of a class was used for each test; Continuous Assessment (CA and Conventional Method (CM were used for each arm. Students were taught during their normal school times for the maximum time of forty minutes thrice a week. There were ten objective questions weekly for each assessment of the students in the CA method for six weeks. The same questions were used throughout for all the schools, done simultaneously for CA. Also, sixty questions at once at the end of the sixth week for CM. Standard deviation and regression equations for the mean values were used in the analysis. The results show that CA could be adjudged to be better off than the CM because of its higher mean values in all the schools than the CM. The higher R2 values of 0.99 and 0.88 revealed stronger correlation between different methods of assessment and the targeted learners. The CA test should be used instead of CM; the CM does not make learners to gain much cognitive knowledge when compare with what CA does to students.

  6. A high-order 3-D spectral-element method for the forward modelling and inversion of gravimetric data—Application to the western Pyrenees

    Science.gov (United States)

    Martin, Roland; Chevrot, Sébastien; Komatitsch, Dimitri; Seoane, Lucia; Spangenberg, Hannah; Wang, Yi; Dufréchou, Grégory; Bonvalot, Sylvain; Bruinsma, Sean

    2017-04-01

    We image the internal density structure of the Pyrenees by inverting gravity data using an a priori density model derived by scaling a Vp model obtained by full waveform inversion of teleseismic P-waves. Gravity anomalies are computed via a 3-D high-order finite-element integration in the same high-order spectral-element grid as the one used to solve the wave equation and thus to obtain the velocity model. The curvature of the Earth and surface topography are taken into account in order to obtain a density model as accurate as possible. The method is validated through comparisons with exact semi-analytical solutions. We show that the spectral-element method drastically accelerates the computations when compared to other more classical methods. Different scaling relations between compressional velocity and density are tested, and the Nafe-Drake relation is the one that leads to the best agreement between computed and observed gravity anomalies. Gravity data inversion is then performed and the results allow us to put more constraints on the density structure of the shallow crust and on the deep architecture of the mountain range.

  7. Development of a dot blot assay with antibodies to recombinant “core” 14-3-3 protein: Evaluation of its usefulness in diagnosis of Creutzfeldt–Jakob disease

    Directory of Open Access Journals (Sweden)

    Sarada Subramanian

    2016-01-01

    Full Text Available Background and Purpose: Definitive diagnosis of Creutzfeldt–Jakob disease (CJD requires demonstration of infective prion protein (PrPSc in brain tissues by immunohistochemistry or immunoblot, making antemortem diagnosis of CJD difficult. The World Health Organization (WHO recommends detection of 14-3-3 protein in cerebrospinal fluid (CSF in cases of dementia, with clinical correlation, as a useful diagnostic marker for CJD, obviating the need for brain biopsy.This facility is currently available in only a few specialized centers in the West and no commercial kit is available for clinical diagnostic use in India. Hence the objective of this study was to develop an in-house sensitive assay for quantitation of 14-3-3 protein and to evaluate its diagnostic potential to detect 14-3-3 proteins in CSF as a biomarker in suspected cases of CJD. Materials and Methods: A minigene expressing the “core” 14-3-3 protein was synthesized by overlapping polymerase chain reaction (PCR and the recombinant protein was produced by employing a bacterial expression system. Polyclonal antibodies raised in rabbit against the purified recombinant protein were used for developing a dot blot assay with avidin-biotin technology for signal amplification and quantitation of 14-3-3 protein in CSF. Results: The results in the present study suggest the diagnostic potential of the dot blot method with about 10-fold difference (P< 0.001 in the CSF levels of 14-3-3 protein between the CJD cases (N= 50 and disease controls (N= 70. The receiver operating characteristic (ROC analysis of the results suggested an optimal cutoff value of 2 ng/mL. Conclusions: We have developed an indigenous, economical, and sensitive dot blot method for the quantitation of 14-3-3 protein in CSF.

  8. Antibody Banding Patterns of the Enzyme-Linked Immunoelectrotransfer Blot and Brain Imaging Findings in Patients With Neurocysticercosis.

    Science.gov (United States)

    Arroyo, Gianfranco; Rodriguez, Silvia; Lescano, Andres G; Alroy, Karen A; Bustos, Javier A; Santivañez, Saul; Gonzales, Isidro; Saavedra, Herbert; Pretell, E Javier; Gonzalez, Armando E; Gilman, Robert H; Tsang, Victor C W; Garcia, Hector H

    2018-01-06

    The enzyme-linked immunoelectrotransfer blot (EITB) assay is the reference serological test for neurocysticercosis (NCC). A positive result on EITB does not always correlate with the presence of active infections in the central nervous system (CNS), and patients with a single viable brain cyst may be EITB negative. Nonetheless, EITB antibody banding patterns appears to be related with the expression of 3 protein families of Taenia solium, and in turn with the characteristics of NCC in the CNS (type, stage, and burden of viable cysts). We evaluated EITB antibody banding patterns and brain imaging findings of 548 NCC cases. Similar banding patterns were grouped into homogeneous classes using latent class analysis. The association between classes and brain imaging findings was assessed. Four classes were identified. Class 1 (patients negative or only positive to the GP50 band, related to the protein family of the same name) was associated with nonviable or single viable parenchymal cysticerci; class 2 (patients positive to bands GP42-39 and GP24, related to the T24-42 protein family, with or without anti-GP50 antibodies) was associated with intraparenchymal viable and nonviable infections; classes 3 and 4 (positive to GP50, GP42-39, and GP24 but also responding to low molecular weight bands GP21, GP18, GP14, and GP13, related to the 8 kDa protein family) were associated with extraparenchymal and intraparenchymal multiple viable cysticerci. EITB antibody banding patterns correlate with brain imaging findings and complement imaging information for the diagnosis of NCC and for staging NCC patients. © The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.

  9. Comparison of the performance in detection of HPV infections between the high-risk HPV genotyping real time PCR and the PCR-reverse dot blot assays.

    Science.gov (United States)

    Zhang, Lahong; Dai, Yibei; Chen, Jiahuan; Hong, Liquan; Liu, Yuhua; Ke, Qiang; Chen, Yiwen; Cai, Chengsong; Liu, Xia; Chen, Zhaojun

    2018-01-01

    A new multiplex real-time PCR assay, the high-risk HPV genotyping real time PCR assay (HR HPV RT-PCR), has been developed to detect 15 high-risk HPV types with respective viral loads. In this report, a total of 684 cervical specimens from women diagnosed with vaginitis were assessed by the HR HPV RT-PCR and the PCR reaction and reverse dot blot (PCR-RDB) assays, using a PCR-sequencing method as a reference standard. A total coincidence of 97.7% between the HR HPV RT PCR and the PCR-RDB assays was determined with a Kappa value of 0.953. The HR HPV RT PCR assay had sensitivity, specificity, and concordance rates (accuracy) of 99.7%, 99.7%, and 99.7%, respectively, as confirmed by PCR-sequencing, while the PCR-RDB assay had respective rates of 98.8%, 97.1%, and 98.0%. The overall rate of HPV infection, determined by PCR-sequencing, in women diagnosed with vaginitis was 49.85%, including 36.26% of single infection and 13.6% of multiple infections. The most common infections among the 15 high-risk HPV types in women diagnosed with vaginitis were HPV-52, HPV-16, and HPV-58, with a total detection rate of 10.23%, 7.75%, and 5.85%, respectively. We conclude that the HR HPV RT PCR assay exhibits better clinical performance than the PCR-RDB assay, and is an ideal alternative method for HPV genotyping. In addition, the HR HPV RT PCR assay provides HPV DNA viral loads, and could serve as a quantitative marker in the diagnosis and treatment of single and multiple HPV infections. © 2017 Wiley Periodicals, Inc.

  10. Discovery of novel poly(ADP-ribose) glycohydrolase inhibitors by a quantitative assay system using dot-blot with anti-poly(ADP-ribose)

    International Nuclear Information System (INIS)

    Okita, Naoyuki; Ashizawa, Daisuke; Ohta, Ryo; Abe, Hideaki; Tanuma, Sei-ichi

    2010-01-01

    Poly(ADP-ribosyl)ation, which is mainly regulated by poly(ADP-ribose) polymerase (PARP) and poly(ADP-ribose) glycohydrolase (PARG), is a unique protein modification involved in cellular responses such as DNA repair and replication. PARG hydrolyzes glycosidic linkages of poly(ADP-ribose) synthesized by PARP and liberates ADP-ribose residues. Recent studies have suggested that inhibitors of PARG are able to be potent anti-cancer drug. In order to discover the potent and specific Inhibitors of PARG, a quantitative and high-throughput screening assay system is required. However, previous PARG assay systems are not appropriate for high-throughput screening because PARG activity is measured by radioactivities of ADP-ribose residues released from radioisotope (RI)-labeled poly(ADP-ribose). In this study, we developed a non-RI and quantitative assay system for PARG activity based on dot-blot assay using anti-poly(ADP-ribose) and nitrocellulose membrane. By our method, the maximum velocity (V max ) and the michaelis constant (k m ) of PARG reaction were 4.46 μM and 128.33 μmol/min/mg, respectively. Furthermore, the IC50 of adenosine diphosphate (hydroxymethyl) pyrrolidinediol (ADP-HPD), known as a non-competitive PARG inhibitor, was 0.66 μM. These kinetics values were similar to those obtained by traditional PARG assays. By using our assay system, we discovered two novel PARG inhibitors that have xanthene scaffold. Thus, our quantitative and convenient method is useful for a high-throughput screening of PARG specific inhibitors.

  11. Buddha philosophy and western psychology.

    Science.gov (United States)

    Aich, Tapas Kumar

    2013-01-01

    Four noble truths as preached by Buddha are that the life is full of suffering (Duhkha), that there is a cause of this suffering (Duhkha-samudaya), it is possible to stop suffering (Duhkha-nirodha), and there is a way to extinguish suffering (Duhkha-nirodha-marga). Eight fold Path (astangika-marga) as advocated by Buddha as a way to extinguish the sufferings are right views, right resolve/aspiration, right speech, right action/conduct, right livelihood, right effort right mindfulness and right concentration. Mid-twentieth century saw the collaborations between many psychoanalysts and Buddhist scholars as a meeting between "two of the most powerful forces" operating in the Western mind. Buddhism and Western Psychology overlap in theory and in practice. Over the last century, experts have written on many commonalities between Buddhism and various branches of modern western psychology like phenomenological psychology, psychoanalytical psychotherapy, humanistic psychology, cognitive psychology and existential psychology. Orientalist Alan Watts wrote 'if we look deeply into such ways of life as Buddhism, we do not find either philosophy or religion as these are understood in the West. We find something more nearly resembling psychotherapy'. Buddha was a unique psychotherapist. His therapeutic methods helped millions of people throughout the centuries. This essay is just an expression of what little the current author has understood on Buddha philosophy and an opportunity to offer his deep tribute to one of the greatest psychotherapists the world has ever produced!

  12. Buddha philosophy and western psychology

    Science.gov (United States)

    Aich, Tapas Kumar

    2013-01-01

    Four noble truths as preached by Buddha are that the life is full of suffering (Duhkha), that there is a cause of this suffering (Duhkha-samudaya), it is possible to stop suffering (Duhkha-nirodha), and there is a way to extinguish suffering (Duhkha-nirodha-marga). Eight fold Path (astangika-marga) as advocated by Buddha as a way to extinguish the sufferings are right views, right resolve/aspiration, right speech, right action/conduct, right livelihood, right effort right mindfulness and right concentration. Mid-twentieth century saw the collaborations between many psychoanalysts and Buddhist scholars as a meeting between “two of the most powerful forces” operating in the Western mind. Buddhism and Western Psychology overlap in theory and in practice. Over the last century, experts have written on many commonalities between Buddhism and various branches of modern western psychology like phenomenological psychology, psychoanalytical psychotherapy, humanistic psychology, cognitive psychology and existential psychology. Orientalist Alan Watts wrote ‘if we look deeply into such ways of life as Buddhism, we do not find either philosophy or religion as these are understood in the West. We find something more nearly resembling psychotherapy’. Buddha was a unique psychotherapist. His therapeutic methods helped millions of people throughout the centuries. This essay is just an expression of what little the current author has understood on Buddha philosophy and an opportunity to offer his deep tribute to one of the greatest psychotherapists the world has ever produced! PMID:23858249

  13. Evaluation of the radioimmunoassay, indirect enzyme linked immunosorbent assay, and dot blot assay for the identification of Xanthomonas campestris pv. phaseoli

    Energy Technology Data Exchange (ETDEWEB)

    Malin, E; Belden, E L; Roth, D

    1985-09-01

    A radioimmunoassay (RIA), an indirect competitive enzyme-linked immunosorbent assay (ELISA), and a dot-blot modification of the ELISA were evaluated for detection and identification of Xanthomonas campestris pv. phaseoli (X. c. pv. phaseoli). RIA and the dot blot tests were specific for X. c. pv. phaseoli; however, significant cross reactions occurred in the indirect competitive ELISA when using anti-X. c. pv. phaseoli antiserum against other closely related bacteria. The sensitivity level of all procedures for X. c. pv. phaseoli was approximately l0/sup 5/ colony forming unitsmL. All procedures were unsatisfactory in reliably detecting low levels of X. c. pv. phaseoli directly from extracts of bean seed. However when used in conjunction with ilution plating the dot blot assay and the RIA would be useful in specifically identifying X. c. pv. phaseoli. The relative merits of these tests for identification of X. c. pv. phaseoli are discussed.

  14. A study of wrecked Dovekies (Alle alle) in the western North Atlantic highlights the importance of using standardized methods to quantify plastic ingestion.

    Science.gov (United States)

    Avery-Gomm, Stephanie; Valliant, Michelle; Schacter, Carley R; Robbins, Katherine F; Liboiron, Max; Daoust, Pierre-Yves; Rios, Lorena M; Jones, Ian L

    2016-12-15

    Quantification of plastic ingestion across a range of seabirds is required to assess the prevalence of plastics in marine food webs. We quantified plastic ingestion in beached Dovekies (Alle alle), following a wreck in Newfoundland, Canada. Of 171 birds, 30.4% had ingested plastic (mean 0.81±0.30 SE pieces per bird, mass 0.005±0.002 SE g per bird). Most plastics were fragments of polyethylene and polypropylene. Surprisingly, 37% were burned or melted, indicating a previously unreported source of ingested plastics (incinerated waste). We found no relationship between plastic ingestion and age, sex or body condition. By comparing our results with a similar nearby study, we illustrate the need for researchers to adopt standardized methods for plastic ingestion studies. We underline the importance of using histological techniques to reliably identify gastric pathologies, and advise caution when inferring population level trends in plastic ingestion from studies of emaciated, wrecked birds. Copyright © 2016 Elsevier Ltd. All rights reserved.

  15. Basin Characterisation by Means of Joint Inversion of Electromagnetic Geophysical Data, Borehole Data and Multivariate Statistical Methods: The Loop Head Peninsula, Western Ireland, Case Study

    Science.gov (United States)

    Campanya, J. L.; Ogaya, X.; Jones, A. G.; Rath, V.; McConnell, B.; Haughton, P.; Prada, M.

    2016-12-01

    The Science Foundation Ireland funded project IRECCSEM project (www.ireccsem.ie) aims to evaluate Ireland's potential for onshore carbon sequestration in saline aquifers by integrating new electromagnetic geophysical data with existing geophysical and geological data. One of the objectives of this component of IRECCSEM is to characterise the subsurface beneath the Loop Head Peninsula (part of Clare Basin, Co. Clare, Ireland), and identify major electrical resistivity structures that can guide an interpretation of the carbon sequestration potential of this area. During the summer of 2014, a magnetotelluric (MT) survey was carried out on the Loop Head Peninsula, and data from a total of 140 sites were acquired, including audio-magnetotelluric (AMT), and broadband magnetotelluric (BBMT). The dataset was used to generate shallow three-dimensional (3-D) electrical resistivity models constraining the subsurface to depths of up to 3.5 km. The three-dimensional (3-D) joint inversions were performed using three different types of electromagnetic data: MT impedance tensor (Z), geomagnetic transfer functions (T), and inter-station horizontal magnetic transfer-functions (H). The interpretation of the results was complemented with second-derivative models of the resulting electrical resistivity models, and a quantitative comparison with borehole data using multivariate statistical methods. Second-derivative models were used to define the main interfaces between the geoelectrical structures, facilitating superior comparison with geological and seismic results, and also reducing the influence of the colour scale when interpreting the results. Specific analysis was performed to compare the extant borehole data with the electrical resistivity model, identifying those structures that are better characterised by the resistivity model. Finally, the electrical resistivity model was also used to propagate some of the physical properties measured in the borehole, when a good relation was

  16. Western Australian natural gas

    International Nuclear Information System (INIS)

    Harman, Frank

    1994-01-01

    Western Australia has 80% of Australia's natural gas resources. These are currently exploited to supply the Western Australian market and LNG to Japan. Growth in the market is dependent on limited prospects for power generation and mineral resource processing. Future exploitation of gas resources will require new export LNG markets and/or the installations of a transcontinental pipeline to eastern Australia. The transcontinental option should only be considered after other options for energy supply in eastern Australia are eliminated. Competition to meet market growth in North-east Asia will be considerable and Australia lacks the policies to underpin future LNG capacity. (author)

  17. The Effectiveness of Western Psychotherapy in treating ...

    African Journals Online (AJOL)

    Background: Psychotherapy has been shown to be effective in the treatment of mental disorders in the western world but viewed as an alien method of treatment to Africans. Aim: To review the literature on the effectiveness of psychotherapy in sub- Saharan Africa. Method: A systematic search of Medline, PsychINFO, ...

  18. ANCA-GBM dot-blot : Evaluation of an assay in the differential diagnosis of patients presenting with rapidly progressive glomerulonephritis

    NARCIS (Netherlands)

    Rutgers, Abraham; Damoiseaux, Jan; Roozendaal, Caroline; Limburg, Pieter C; Stegeman, Coen A; Tervaert, Jan Willem Cohen

    Rapidly progressive glomerulonephritis (RPGN) is characterized by rapid and progressive loss of renal function and the presence of crescentic glomerulonephritis (CGN). Early diagnosis and appropriate treatment is mandatory to prevent death and/or renal failure. We have evaluated an ANCA-GBM dot-blot

  19. Imams in Western Europe

    DEFF Research Database (Denmark)

    As European Muslims and Muslims in the Middle East diverge, imams in Europe have emerged as major agents of religious authority who shape Islam’s presence in Western societies. This volume examines the theoretical and practical questions concerning the evolving role of imams in Europe. To what...

  20. Western Military Culture and Counterinsurgency:

    African Journals Online (AJOL)

    francois

    with a threat both abroad and within their homeland societies. Civilians fulfil a ..... we have now with the use of force and forces is their persistent structuring ... advanced equipment remains the main feature of Western military culture. Western.

  1. Suicide and Western culture.

    Science.gov (United States)

    Pridmore, Saxby; McArthur, Milford

    2009-02-01

    The aim of this paper is to examine the cultural roots and transmission of Western suicide and suicidal behaviour. We explored a period of antiquity (mythical Greece-61 CE) and selected accounts of 10 prominent suicides. The precipitating circumstances were tabulated and an assessment made of the most likely attendant emotions. The same process was followed for a recent period (1994-2008), from which 10 suicides were identified. The precipitating circumstances and the attendant emotions were compared. These circumstances and emotions were then compared to statements commonly encountered in clinical practice from people demonstrating suicidal behaviour. Finally, we looked for evidence that these stories (and the response models) had entered Western culture. Precipitating circumstances, loss of a loved one, actual or imminent execution or imprisonment, other losses and public disgrace, and the negative emotions of shame, guilt, fear, anger, grief and sorrow were common to both historical periods. These circumstances and emotions are similar to those commonly expressed by people who have demonstrated suicidal behaviour. There was a clear record (literature, visual arts) of these stories forming part of our cultural heritage. Models of maladaptive responses to certain adverse circumstances are part of Western culture. Suicide as a response to certain circumstances and negative emotions can be traced back more than 2000 years. Cultural change will be necessary to minimize suicide.

  2. Internal globalization of Western Balkan

    Directory of Open Access Journals (Sweden)

    Vukotić Veselin

    2006-01-01

    Full Text Available What are potential and real effects of the globalization process on the economic connection between Western Balkan countries? What is the crucial change in relations between Western Balkan countries and its economies inexorably brought by globalization? What are the elements of political economy of Western Balkan globalization? What are reflections of the conflict between political and economic areas of Western Balkan? These are some of the issues discuses in this paper.

  3. MStern Blotting-High Throughput Polyvinylidene Fluoride (PVDF) Membrane-Based Proteomic Sample Preparation for 96-Well Plates.

    Science.gov (United States)

    Berger, Sebastian T; Ahmed, Saima; Muntel, Jan; Cuevas Polo, Nerea; Bachur, Richard; Kentsis, Alex; Steen, Judith; Steen, Hanno

    2015-10-01

    We describe a 96-well plate compatible membrane-based proteomic sample processing method, which enables the complete processing of 96 samples (or multiples thereof) within a single workday. This method uses a large-pore hydrophobic PVDF membrane that efficiently adsorbs proteins, resulting in fast liquid transfer through the membrane and significantly reduced sample processing times. Low liquid transfer speeds have prevented the useful 96-well plate implementation of FASP as a widely used membrane-based proteomic sample processing method. We validated our approach on whole-cell lysate and urine and cerebrospinal fluid as clinically relevant body fluids. Without compromising peptide and protein identification, our method uses a vacuum manifold and circumvents the need for digest desalting, making our processing method compatible with standard liquid handling robots. In summary, our new method maintains the strengths of FASP and simultaneously overcomes one of the major limitations of FASP without compromising protein identification and quantification. © 2015 by The American Society for Biochemistry and Molecular Biology, Inc.

  4. International students' experience of a western medical school: a mixed methods study exploring the early years in the context of cultural and social adjustment compared to students from the host country.

    Science.gov (United States)

    McGarvey, A; Brugha, R; Conroy, R M; Clarke, E; Byrne, E

    2015-07-02

    Few studies have addressed the challenges associated with international students as they adapt to studying medicine in a new host country. Higher level institutions have increasing numbers of international students commencing programmes. This paper explores the experiences of a cohort of students in the early years of medical school in Ireland, where a considerable cohort are from an international background. A mixed exploratory sequential study design was carried out with medical students in the preclinical component of a five year undergraduate programme. Data for the qualitative phase was collected through 29 semi-structured interviews using the peer interview method. Thematic analysis from this phase was incorporated to develop an online questionnaire combined with components of the Student Adaptation to College Questionnaire and Student Integration Questionnaire. First year students were anonymously surveyed online. The Mokken Scaling procedure was used to investigate the students' experiences, both positive and negative. Three main themes are identified; social adjustment, social alienation and cultural alienation. The response rate for the survey was 49% (467 Respondents). The Mokken Scaling method identified the following scales (i) Positive experience of student life; (ii) Social alienation, which comprised of negative items about feeling lonely, not fitting in, being homesick and (iii) Cultural alienation, which included the items of being uncomfortable around cultural norms of dress and contact between the sexes. With the threshold set to H = 0.4. Subscales of the positive experiences of student life scale are explored further. Overall student adjustment to a western third level college was good. Students from regions where cultural distance is greatest reported more difficulties in adjusting. Students from these regions also demonstrate very good adaptation. Some students from the host country and more similar cultural backgrounds were also

  5. Identification of pathogenic Nocardia species by reverse line blot hybridization targeting the 16S rRNA and 16S-23S rRNA gene spacer regions.

    Science.gov (United States)

    Xiao, Meng; Kong, Fanrong; Sorrell, Tania C; Cao, Yongyan; Lee, Ok Cha; Liu, Ying; Sintchenko, Vitali; Chen, Sharon C A

    2010-02-01

    Although 16S rRNA gene sequence analysis is employed most often for the definitive identification of Nocardia species, alternate molecular methods and polymorphisms in other gene targets have also enabled species determinations. We evaluated a combined Nocardia PCR-based reverse line blot (RLB) hybridization assay based on 16S and 16S-23S rRNA gene spacer region polymorphisms to identify 12 American Type Culture Collection and 123 clinical Nocardia isolates representing 14 species; results were compared with results from 16S rRNA gene sequencing. Thirteen 16S rRNA gene-based (two group-specific and 11 species-specific) and five 16S-23S spacer-targeted (two taxon-specific and three species-specific) probes were utilized. 16S rRNA gene-based probes correctly identified 124 of 135 isolates (sensitivity, 92%) but were unable to identify Nocardia paucivorans strains (n = 10 strains) and a Nocardia asteroides isolate with a novel 16S rRNA gene sequence. Nocardia farcinica and Nocardia cyriacigeorgica strains were identified by the sequential use of an N. farcinica-"negative" probe and a combined N. farcinica/N. cyriacigeorgica probe. The assay specificity was high (99%) except for weak cross-reactivity between the Nocardia brasiliensis probe with the Nocardia thailandica DNA product; however, cross-hybridization with closely related nontarget species may occur. The incorporation of 16S-23S rRNA gene spacer-based probes enabled the identification of all N. paucivorans strains. The overall sensitivity using both probe sets was >99%. Both N. farcinica-specific 16S-23S rRNA gene spacer-directed probes were required to identify all N. farcinica stains by using this probe set. The study demonstrates the utility of a combined PCR/RLB assay for the identification of clinically relevant Nocardia species and its potential for studying subtypes of N. farcinica. Where species assignment is ambiguous or not possible, 16S rRNA gene sequencing is recommended.

  6. Detection of human papilloma virus 16 and 18 DNA sequences by southern blot hybridization in oral leukoplakia and squamous cell carcinoma.

    Science.gov (United States)

    Khanna, Rahul; Rao, G R K; Tiwary, S K; Rai, Ashish; Khanna, Seema; Khanna, A K

    2009-04-01

    The etiopathological role of human papilloma virus (HPV) in the causation of oral cancer is till a subject of speculation. We used the technique of Southern blot hybridization to detect the presence of HPV types 16 & 18 in biopsy specimens from oral cancer and leukoplakia patients as well as normal oral mucosal biopsies. The prevalence of either HPV type 16 or 18 was found in 64.5% (29/45) of oral cancer, 40%(12/30) of leukoplakia and 20%(9/45) of normal oral mucosal biopsies. No association could be demonstrated between tobacco usage habits or a history of genital warts with HPV prevalence. A significant finding was that none of the oral cancer patients were negative for both: a history of tobacco usage as well as presence of HPV infection, on Southern blot hybridization.

  7. Hepatitis e virus: Western Cape, South Africa

    NARCIS (Netherlands)

    R.G. Madden (Richie); Wallace, S. (Sebastian); M. Sonderup; Korsman, S. (Stephen); Chivese, T. (Tawanda); Gavine, B. (Bronwyn); Edem, A. (Aniefiok); Govender, R. (Roxy); English, N. (Nathan); Kaiyamo, C. (Christy); Lutchman, O. (Odelia); A.A. Eijck (Annemiek); S.D. Pas (Suzan); Webb, G.W. (Glynn W); Palmer, J. (Joanne); Goddard, E. (Elizabeth); Wasserman, S. (Sean); H.R. Dalton (Harry); C.W. Spearman

    2016-01-01

    textabstractAIM To conduct a prospective assessment of anti-hepatitis E virus (HEV) IgG seroprevalence in the Western Cape Province of South Africa in conjunction with evaluating risk factors for exposure. METHODS Consenting participants attending clinics and wards of Groote Schuur, Red Cross

  8. Establishment and application of a modified membrane-blot assay for Rhizomucor miehei lipases aimed at improving their methanol tolerance and thermostability.

    Science.gov (United States)

    He, Dong; Luo, Wen; Wang, Zhiyuan; Lv, Pengmei; Yuan, Zhenhong; Huang, Shaowei; Xv, Jingliang

    2017-07-01

    Directed evolution has been proved an effective way to improve the stability of proteins, but high throughput screening assays for directed evolution with simultaneous improvement of two or more properties are still rare. In this study, we aimed to establish a membrane-blot assay for use in the high-throughput screening of Rhizomucor miehei lipases (RMLs). With the assistance of the membrane-blot screening assay, a mutant E47K named G10 that showed improved thermal stability was detected in the first round of error-prone PCR. Using G10 as the parent, two variants G10-11 and G10-20 that showed improved thermal stability and methanol tolerance without loss of activity compared to the wild type RML were obtained. The T 50 60 -value of G10-11 and G10-20 increased by 12°C and 6.5°C, respectively. After incubation for 1h, the remaining residual activity of G10-11 and G10-20 was 63.45% and 74.33%, respectively, in 50% methanol, and 15.98% and 30.22%, respectively, in 80% methanol. Thus, we successfully developed a membrane-blot assay that could be used for the high-throughput screening of RMLs with improved thermostability and methanol tolerance. Based on our findings, we believe that our newly developed membrane-blot assay will have potential applications in directed evolution in the future. Copyright © 2017 Elsevier Inc. All rights reserved.

  9. Nuclear energy in Western Europe

    International Nuclear Information System (INIS)

    Loennroth, M.; Walker, W.

    1984-01-01

    This is an overview of nuclear energy in Western Europe, as seen by two Western Europeans, attempting to place the topic into the context not only of energy supply but also of industrial relations, institutional structure, and sociocultural factors. Although its main focus is Western Europe, it is sometimes necessary to glance at the wider context, in particular the industrial relations with the United States and Japan. Export markets are also considered, in particular, in the Pacific. The paper does not, however, deal with nonproliferation policies and the possible difference of opinion within Western Europe and between Western Europe and other regions over this topic. (author)

  10. Rethinking Education Research methodologies-Dominance of Western perspectives?

    Directory of Open Access Journals (Sweden)

    Yu Sing Ong

    2016-11-01

    Full Text Available This paper examines the different approaches and methods in comparative education research. It begins with a broad overview of the influences of Western philosophies and how Western education researchers approach social situations. Its differences with Chinese philosophies were highlighted with the concept of dao and Confucianism. More than just an epistemological discussion, this paper argues that Western researchers’ biasness towards Asian researchers may be due to their failure or lack of experience in making the connections with Asian social systems. It attempts to provide some plausible explanations of the prevalence of biasness by peer reviewers of Western journals.

  11. A novel method for in Situ detection of hydrolyzable casein fragments in a cheese matrix by antibody phage display technique and CLSM

    DEFF Research Database (Denmark)

    Duan, Zhi; Brüggemann, Dagmar Adeline; Siegumfeldt, Henrik

    2009-01-01

    three small synthetic peptides of the alpha(s1)-casein sequence. These peptides traverse enzymatic cleavage sites of casein during cheese ripening. The specificity of the generated anti-peptide antibodies was determined by ELISA and Western blot. Finally, an immunofluorescent labeling protocol......A novel method to monitor in situ hydrolyzable casein fragments during cheese ripening by using immunofluorescent labeling and confocal laser scanning microscopy (CLSM) was developed. Monoclonal single chain variable fragments of antibody (scFvs) were generated by antibody phage display toward...

  12. Limitations of the colloidal silica method in mapping the endothelial plasma membrane proteome of the mouse heart.

    Science.gov (United States)

    Arjunan, Selvam; Reinartz, Michael; Emde, Barbara; Zanger, Klaus; Schrader, Jürgen

    2009-01-01

    The endothelial cell (EC) membrane is an important interface, which plays a crucial role in signal transduction. Our aim was to selectively purify luminal EC membrane proteins from the coronary vasculature of the isolated perfused mouse heart and analyze its composition with mass spectrometry (MS). To specifically label coronary ECs in the intact heart, the colloidal silica method was applied, which is based on the binding of positively charged colloidal silica to the surface of EC membranes. Transmission electron microscopy revealed the specific labeling of ECs of macro and microvessels. Two different methods of tissue homogenization (Teflon pestle and ultra blade) together with density centrifugation were used for membrane protein enrichment. Enrichment and purity was controlled by Western blot analysis using the EC-specific protein caveolin 1 and various intracellular marker proteins. The ultra blade method resulted in a tenfold enrichment of caveolin 1, while there was negligible contamination as judged by Western blot. However, protein yield was low and required pooling of ten hearts for MS. When enriched endothelial membrane proteins were digested with trypsin and analyzed by LC-MS, a total of 56 proteins could be identified, of which only 12 were membrane proteins. We conclude that coronary endothelial membranes can be conveniently labeled with colloidal silica. However, due to the ionic nature of interaction of colloidal silica with the EC membrane the shear rate required for cardiac homogenization resulted in a substantial loss of specificity.

  13. Evaluation of Two Commercial Systems for Automated Processing, Reading, and Interpretation of Lyme Borreliosis Western Blots▿

    Science.gov (United States)

    Binnicker, M. J.; Jespersen, D. J.; Harring, J. A.; Rollins, L. O.; Bryant, S. C.; Beito, E. M.

    2008-01-01

    The diagnosis of Lyme borreliosis (LB) is commonly made by serologic testing with Western blot (WB) analysis serving as an important supplemental assay. Although specific, the interpretation of WBs for diagnosis of LB (i.e., Lyme WBs) is subjective, with considerable variability in results. In addition, the processing, reading, and interpretation of Lyme WBs are laborious and time-consuming procedures. With the need for rapid processing and more objective interpretation of Lyme WBs, we evaluated the performances of two automated interpretive systems, TrinBlot/BLOTrix (Trinity Biotech, Carlsbad, CA) and BeeBlot/ViraScan (Viramed Biotech AG, Munich, Germany), using 518 serum specimens submitted to our laboratory for Lyme WB analysis. The results of routine testing with visual interpretation were compared to those obtained by BLOTrix analysis of MarBlot immunoglobulin M (IgM) and IgG and by ViraScan analysis of ViraBlot and ViraStripe IgM and IgG assays. BLOTrix analysis demonstrated an agreement of 84.7% for IgM and 87.3% for IgG compared to visual reading and interpretation. ViraScan analysis of the ViraBlot assays demonstrated agreements of 85.7% for IgM and 94.2% for IgG, while ViraScan analysis of the ViraStripe IgM and IgG assays showed agreements of 87.1 and 93.1%, respectively. Testing by the automated systems yielded an average time savings of 64 min/run compared to processing, reading, and interpretation by our current procedure. Our findings demonstrated that automated processing and interpretive systems yield results comparable to those of visual interpretation, while reducing the subjectivity and time required for Lyme WB analysis. PMID:18463211

  14. Lake Carnegie, Western Australia

    Science.gov (United States)

    2002-01-01

    Ephemeral Lake Carnegie, in Western Australia, fills with water only during periods of significant rainfall. In dry years, it is reduced to a muddy marsh. This image was acquired by Landsat 7's Enhanced Thematic Mapper plus (ETM+) sensor on May 19, 1999. This is a false-color composite image made using shortwave infrared, infrared, and red wavelengths. The image has also been sharpened using the sensor's panchromatic band. Image provided by the USGS EROS Data Center Satellite Systems Branch. This image is part of the ongoing Landsat Earth as Art series.

  15. Western Canada uranium perspective

    International Nuclear Information System (INIS)

    Lloyd, R.E.

    1984-01-01

    The current situation in the exploration for uranium in British Columbia, the Yukon, the Northwest Territories, and Saskatchewan is reviewed. A moratorium on exploration has been in effect in British Columbia since 1980; it is due to expire in 1987. Only the Blizzard deposit appears to have any economic potential. The Lone Gull discovery in the Thelon Basin of the Northwest Territories has proven reserves of more than 35 million pounds U 3 O 8 grading 0.4%. Potentially prospective areas of the northern Thelon Basin lie within a game sanctuary and cannot be explored. Exploration activity in Saskatchewan continues to decline from the peak in 1980. Three major deposits - Cluff Lake, Rabbit Lake and Key Lake - are in production. By 1985 Saskatchewan will produce 58% of Canada's uranium, and over 13% of the western world's output. (L.L.) (3 figs, 2 tabs.)

  16. Contribution of dot-blot assay to the diagnosis and management of myositis: a three-year practice at a university hospital centre.

    Science.gov (United States)

    Martel, Clothilde; Vignaud, Guillaume; Liozon, Eric; Magy, Laurent; Gallouedec, Gael; Ly, Kim; Bezanahary, Holly; Cypierre, Anne; Lapébie, François-Xavier; Palat, Sylvain; Gondran, Guillaume; Jauberteau, Marie-Odile; Fauchais, Anne-Laure

    2016-01-01

    Idiopathic inflammatory myopathies (IIM) are heterogeneous autoimmune diseases with wide clinical spectrum that may lead to delayed diagnosis. The aim of this study was to examine the impact of IIM-specific dot-blot assay on diagnostic process of patients presenting with muscular or systemic symptoms evocating of IIM. We collected all the prescriptions of an IIM specific dot-blot assay (8 autoantigens including Jo-1, PL-7, PL-12, SRP, Mi-2, Ku, PM/Scl and Scl-70) over a 38-month period. 316 myositis dot-blot assays (MSD) were performed in 274 patients (156 women, mean age 53±10.6 years) referring for muscular and/or systemic symptoms suggesting IIM. The timing of dot prescription through the diagnostic process was highly variable: without (35%), concomitantly (16%) or after electromyographic studies (35%). Fifty-nine patients (22%) had IIM according to Bohan and Peter's criteria. Among them, 29 (49%) had positive dot (8 Jo-1, 6 PM-Scl, 5 PL-12, 5 SRP, 2 Mi-2, 2 PL-7 and 1 Ku). Various other diagnoses were performed including 35 autoimmune disease or granulomatosis (12%), 19 inflammatory rheumatic disease (7%), 16 non inflammatory muscular disorders (6%), 10 drug-induced myalgia (4%), 11 infectious myositis (4%). Except 11 borderline SRP results and one transient PM-Scl, MSD was positive only in one case of IIM. Dot allowed clinicians to correct diagnosis in 4 cases and improved the diagnosis of IIM subtypes in 4 cases. This study reflects the interest of myositis dot in the rapid diagnosis process of patients with non-specific muscular symptoms leading to various diagnoses including IIM.

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

    International Nuclear Information System (INIS)

    Lucher, L.A.; Lego, T.

    1989-01-01

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

  18. “They just scraped off the calluses”: a mixed methods exploration of foot care access and provision for people with rheumatoid arthritis in south-western Sydney, Australia

    Science.gov (United States)

    2013-01-01

    Background There is little indication that foot health services in Australia are meeting modern day recommendations for Rheumatoid Arthritis (RA) patients. The overall objective of this study was to explore the current state of foot health services for patients with RA with an emphasis on identifying barriers to the receipt of appropriate foot care in South-West Sydney, New South Wales, Australia. Methods A mixed (quantitative and qualitative) approach was adopted. Indications for appropriate access to foot care were determined by comparing the foot health, disease and socio-demographic characteristics of patients with unmet foot care demands, foot care users and patients with no demands for foot care. Perceptions of provision of, and access to, foot care were explored by conducting telephone-based interviews using an interpretative phenomenology approach with thematic analysis. Results Twenty-nine participants took part in the cross-sectional quantitative research study design, and 12 participants took part in the interpretative phenomenological approach (qualitative study). Foot care access appeared to be driven predominantly by the presence of rearfoot deformity, which was significantly worse amongst participants in the foot care user group (p = 0.02). Five main themes emerged from the qualitative data: 1) impact of disease-related foot symptoms, 2) footwear difficulties, 3) medical/rheumatology encounters, 4) foot and podiatry care access and experiences, and 5) financial hardship. Conclusions Foot care provision does not appear to be driven by appropriate foot health characteristics such as foot pain or foot-related disability. There may be significant shortfalls in footwear and foot care access and provision in Greater Western Sydney. Several barriers to adequate foot care access and provision were identified and further efforts are required to improve access to and the quality of foot care for people who have RA. Integration of podiatry services within

  19. Production of hybrids between western gray wolves and western coyotes.

    Directory of Open Access Journals (Sweden)

    L David Mech

    Full Text Available Using artificial insemination we attempted to produce hybrids between captive, male, western, gray wolves (Canis lupus and female, western coyotes (Canis latrans to determine whether their gametes would be compatible and the coyotes could produce and nurture offspring. The results contribute new information to an ongoing controversy over whether the eastern wolf (Canis lycaon is a valid unique species that could be subject to the U. S. Endangered Species Act. Attempts with transcervically deposited wolf semen into nine coyotes over two breeding seasons yielded three coyote pregnancies. One coyote ate her pups, another produced a resorbed fetus and a dead fetus by C-section, and the third produced seven hybrids, six of which survived. These results show that, although it might be unlikely for male western wolves to successfully produce offspring with female western coyotes under natural conditions, western-gray-wolf sperm are compatible with western-coyote ova and that at least one coyote could produce and nurture hybrid offspring. This finding in turn demonstrates that gamete incompatibility would not have prevented western, gray wolves from inseminating western coyotes and thus producing hybrids with coyote mtDNA, a claim that counters the view that the eastern wolf is a separate species. However, some of the difficulties experienced by the other inseminated coyotes tend to temper that finding and suggest that more experimentation is needed, including determining the behavioral and physical compatibility of western gray wolves copulating with western coyotes. Thus although our study adds new information to the controversy, it does not settle it. Further study is needed to determine whether the putative Canis lycaon is indeed a unique species.

  20. Quantitative analysis of the IgG and IgG subclass immune responses to chromosomal Pseudomonas aeruginosa beta-lactamase in serum from patients with cystic fibrosis by western blotting and laser scanning densitometry

    DEFF Research Database (Denmark)

    Petersen, T D; Ciofu, O; Pressler, T

    1996-01-01

    BACKGROUND: Antibodies against chromosomal beta-lactamase of Pseudomonas aeruginosa (a beta ab) are markers of the development of resistance of P aeruginosa to beta-lactam antibiotics in patients with cystic fibrosis and chronic lung infection. The role of these antibodies in patients with chronic...... of the chronic infection the a beta ab titres were higher in patients with good lung function than in those with poor lung function. CONCLUSIONS: The association of a weak IgG3 and a strong IgG4 a beta ab response suggests that the contribution of a beta ab antibodies to lung diseases mediated by immune...... complexes might be less important than other antipseudomonal antibodies. A beneficial neutralising effect of the a beta ab antibodies on the antibiotic destroying enzymes may be an additional factor....

  1. Analyzing Protein Changes in Guinea Pig Tissue Lysates Using Non-guinea Pig Specific Antibodies: Procedures for Western Blotting and Examples Using 16 Individual Antibodies for Common CNS Proteins

    National Research Council Canada - National Science Library

    Johnson, Erik A; Daugherty, Kelly S

    2006-01-01

    ... behavioral and protein changes due to the absence of guinea pig-specific antibodies. We have developed a procedure to determine the specificity of commercially available, non-guinea pig-specific antibodies in guinea pig lysates...

  2. Analyzing Protein Changes in Guinea Pig Tissue Lysates Using Non-guinea Pig Specific Antibodies: Procedures for Western Blotting and Examples Using 16 Individual Antibodies for Common CNS Proteins

    Science.gov (United States)

    2006-05-01

    guinea pig model does present a significant problem...trying to correlate behavioral and protein changes due to the absence of guinea pig -specific antibodies. We...have developed a procedure to determine the specificity of commercially available, non- guinea pig -specific antibodies in guinea pig lysates.

  3. Demonstration of functional low-density lipoprotein receptors by protein blotting in fibroblasts from a subject with homozygous receptor-negative familial hypercholesterolemia

    International Nuclear Information System (INIS)

    Semenkovich, C.F.; Ostlund, R.E. Jr.; Yang, J.; Reaban, M.E.

    1985-01-01

    We report the detection of low-density lipoprotein (LDL) receptors by the technique of receptor blotting in fibroblasts from a patient with homozygous familial hypercholesterolemia (FHC) previously classified as ''receptor negative.'' Solubilized receptors were electrophoresed, transferred to nitrocellulose paper, treated with LDL followed by radiolabeled antibody to LDL, and visualized by autoradiography. GM 2000 FHC fibroblasts revealed LDL receptors with an apparent molecular weight of approximately 140,000, the same as in normal cells. LDL receptor activity by blotting in GM 2000 cells was greatly diminished in comparison with normal cells, but was calcium dependent. Receptor activity was also detectable by conventional monolayer binding and degradation assays. Thus, GM 2000 cells have profoundly diminished LDL receptor activity, but retain the genetic capacity to make LDL receptor material of normal molecular weight that is capable of binding LDL. Previous studies have demonstrated the presence of trace amounts of immunoreactive LDL receptor protein in fibroblasts from some receptor-negative FHC homozygotes. These studies are extended by demonstrating the ability of this material to bind LDL

  4. Application of FTA sample collection and DNA purification system on the determination of CTG trinucleotide repeat size by PCR-based Southern blotting.

    Science.gov (United States)

    Hsiao, K M; Lin, H M; Pan, H; Li, T C; Chen, S S; Jou, S B; Chiu, Y L; Wu, M F; Lin, C C; Li, S Y

    1999-01-01

    Myotonic dystrophy (DM) is caused by a CTG trinucleotide expansion mutation at exon 15 of the myotonic dystrophy protein kinase gene. The clinical severity of this disease correlates with the length of the CTG trinucleotide repeats. Determination of the CTG repeat length has been primarily relied on by Southern blot analysis of restriction enzyme-digested genomic DNA. The development of PCR-based Southern blotting methodology provides a much more sensitive and simpler protocol for DM diagnosis. However, the quality of the template and the high (G+C) ratio of the amplified region hamper the use of PCR on the diagnosis of DM. A modified PCR protocol to amplify different lengths of CTG repeat region using various concentrations of 7deaza-dGTP has been reported (1). Here we describe a procedure including sample collection, DNA purification, and PCR analysis of CTG repeat length without using 7-deaza-dGTP. This protocol is very sensitive and convenient because only a small number of nucleate cells are needed for detection of CTG expansion. Therefore, it could be very useful in clinical and prenatal diagnosis as well as in prevalence study of DM.

  5. Method

    Directory of Open Access Journals (Sweden)

    Ling Fiona W.M.

    2017-01-01

    Full Text Available Rapid prototyping of microchannel gain lots of attention from researchers along with the rapid development of microfluidic technology. The conventional methods carried few disadvantages such as high cost, time consuming, required high operating pressure and temperature and involve expertise in operating the equipment. In this work, new method adapting xurography method is introduced to replace the conventional method of fabrication of microchannels. The novelty in this study is replacing the adhesion film with clear plastic film which was used to cut the design of the microchannel as the material is more suitable for fabricating more complex microchannel design. The microchannel was then mold using polymethyldisiloxane (PDMS and bonded with a clean glass to produce a close microchannel. The microchannel produced had a clean edge indicating good master mold was produced using the cutting plotter and the bonding between the PDMS and glass was good where no leakage was observed. The materials used in this method is cheap and the total time consumed is less than 5 hours where this method is suitable for rapid prototyping of microchannel.

  6. Competitive edge of western coal

    International Nuclear Information System (INIS)

    Keith, R.D.

    1990-01-01

    This paper expresses views on the competitive advantages of one of the nation's most remarkable energy resources--Western coal. It covers utilization of Western coal, and its advantages. The Arkansas Power and Light Company and its demand for coal are also covered

  7. Use of capillary Western immunoassay (Wes) for quantification of dystrophin levels in skeletal muscle of healthy controls and individuals with Becker and Duchenne muscular dystrophy.

    Science.gov (United States)

    Beekman, Chantal; Janson, Anneke A; Baghat, Aabed; van Deutekom, Judith C; Datson, Nicole A

    2018-01-01

    Duchenne muscular dystrophy (DMD) is a neuromuscular disease characterized by progressive weakness of the skeletal and cardiac muscles. This X-linked disorder is caused by open reading frame disrupting mutations in the DMD gene, resulting in strong reduction or complete absence of dystrophin protein. In order to use dystrophin as a supportive or even surrogate biomarker in clinical studies on investigational drugs aiming at correcting the primary cause of the disease, the ability to reliably quantify dystrophin expression in muscle biopsies of DMD patients pre- and post-treatment is essential. Here we demonstrate the application of the ProteinSimple capillary immunoassay (Wes) method, a gel- and blot-free method requiring less sample, antibody and time to run than conventional Western blot assay. We optimized dystrophin quantification by Wes using 2 different antibodies and found it to be highly sensitive, reproducible and quantitative over a large dynamic range. Using a healthy control muscle sample as a reference and α-actinin as a protein loading/muscle content control, a panel of skeletal muscle samples consisting of 31 healthy controls, 25 Becker Muscle dystrophy (BMD) and 17 DMD samples was subjected to Wes analysis. In healthy controls dystrophin levels varied 3 to 5-fold between the highest and lowest muscle samples, with the reference sample representing the average of all 31 samples. In BMD muscle samples dystrophin levels ranged from 10% to 90%, with an average of 33% of the healthy muscle average, while for the DMD samples the average dystrophin level was 1.3%, ranging from 0.7% to 7% of the healthy muscle average. In conclusion, Wes is a suitable, efficient and reliable method for quantification of dystrophin expression as a biomarker in DMD clinical drug development.

  8. Globalisation and western music historiography

    Directory of Open Access Journals (Sweden)

    Romanou Katy

    2015-01-01

    Full Text Available Globalisation of musicology and music history aims to fuse the divisions created during Western music’s acme, and is referred to as “post-European historical thinking”. Therefore, “post” and “pre” European historical thinking have much in common. One aspect of this process of fragmentation was that music history was separated from theory and that Western Music Histories succeeded General Music Histories (a development described in some detail in the article. Connecting global music history with “post-European” historical thinking is one among numerous indications of Western awareness that European culture has reached some sort of a terminal phase. Concurrently, countries that have been developing by following Western Europe as a prototype, are leading today some past phase of Western development, which, with the ideas of cultural relativism prevailing, are not considered inferior.

  9. Western Nuclear Science Alliance

    International Nuclear Information System (INIS)

    Reese, Steve; Miller, George; Frantz, Stephen; Beller, Denis; Morse, Ed; Krahenbuhl, Melinda; Flocchini, Bob; Elliston, Jim

    2010-01-01

    The Western Nuclear Science Alliance (WNSA) was formed at Oregon State University (OSU) under the DOE Innovations in Nuclear Infrastructure and Education (INIE) program in 2002. The primary objective of the INIE program is to strengthen nuclear science and engineering programs at the member institutions and to address the long term goal of the University Reactor Infrastructure and Education Assistance Program. WNSA has been very effective in meeting these goals. The infrastructure at several of the WNSA university nuclear reactors has been upgraded significantly, as have classroom and laboratory facilities for Nuclear Engineering, Health Physics, and Radiochemistry students and faculty. Major nuclear-related education programs have been inaugurated, including considerable assistance by WNSA universities to other university nuclear programs. Research has also been enhanced under WNSA, as has outreach to pre-college and college students and faculty. The INIE program under WNSA has been an exceptional boost to the nuclear programs at the eight funded WNSA universities. In subsequent years under INIE these programs have expanded even further in terms of new research facilities, research reactor renovations, expanded educational opportunities, and extended cooperation and collaboration between universities, national laboratories, and nuclear utilities.

  10. Eastern and western happiness in work behavior

    OpenAIRE

    Jaroslava Kubátová

    2017-01-01

    The purpose of this article is to clarify what motivates East Asian work behavior. The research question is: How can a Western manager better understand work behavior and motivation in East Asian cultures? Knowledge about national culture, motivation and the concept of happiness are connected via deductive and comparative methods, while pointing out their new connections and relations. We argue that work motivation in East Asian cultures can be explained using the self-concept-based motivatio...

  11. Kan du blot blive buddreng..

    DEFF Research Database (Denmark)

    Henriksen, Carsten

    2011-01-01

    ADHD? Adfærdsvanskelig? Diagnoserne står i kø, når vi i dag skal forklare, hvorfor Emil ikke kan følge med i skolen. Men startskuddet til den videnskabelige opdeling af de børn, der ikke følger flertallets læringskurve, lød allerede for 100 år siden.......ADHD? Adfærdsvanskelig? Diagnoserne står i kø, når vi i dag skal forklare, hvorfor Emil ikke kan følge med i skolen. Men startskuddet til den videnskabelige opdeling af de børn, der ikke følger flertallets læringskurve, lød allerede for 100 år siden....

  12. method

    Directory of Open Access Journals (Sweden)

    L. M. Kimball

    2002-01-01

    Full Text Available This paper presents an interior point algorithm to solve the multiperiod hydrothermal economic dispatch (HTED. The multiperiod HTED is a large scale nonlinear programming problem. Various optimization methods have been applied to the multiperiod HTED, but most neglect important network characteristics or require decomposition into thermal and hydro subproblems. The algorithm described here exploits the special bordered block diagonal structure and sparsity of the Newton system for the first order necessary conditions to result in a fast efficient algorithm that can account for all network aspects. Applying this new algorithm challenges a conventional method for the use of available hydro resources known as the peak shaving heuristic.

  13. Poland, a workforce in transition: Exploring leadership styles and effectiveness of Polish vs. Western expatriate managers

    OpenAIRE

    Eisenberg, Jacob; Pieczonka, Artur; Eisenring, Martin; Mironski, Jacek

    2015-01-01

    Given the cultural differences between Western Europe and Poland, differences are expected in leadership styles and behaviours between Polish and Western managers. Our study explored Polish employees’ perceptions and attitudes toward expatriate Western versus local managers. The main method was surveying Polish employees working under Western managers in three mid-size companies. We supplemented the survey with in-depth interviews with five West European middle managers working in Poland. We ...

  14. Decommissioning in western Europe

    International Nuclear Information System (INIS)

    Lundqvist, K.

    1999-12-01

    This report gives an overview of the situation in Western Europe. The original aim was to focus on organisational and human issues with regard to nuclear reactor decommissioning, but very few articles were found. This is in sharp contrast to the substantial literature on technical issues. While most of the reports on decommissioning have a technical focus, several provide information on regulatory issues, strategies and 'state of the art'. The importance of the human and organizational perspective is however discovered, when reading between the lines of the technical publications, and especially when project managers summarize lessons learned. The results are to a large extent based on studies of articles and reports, mainly collected from the INIS database. Decommissioning of nuclear facilities started already in the sixties, but then mainly research and experimental facilities were concerned. Until now about 70 reactors have been shutdown world-wide. Over the years there have been plenty of conferences for exchanging experiences mostly about technical matters. Waste Management is a big issue. In the 2000s there will be a wave of decommissioning when an increasing amount of reactors will reach the end of their calculated lifetime (40 years, a figure now being challenged by both life-extension and pre-shutdown projects). Several reactors have been shut-down for economical reasons. Shutdown and decommissioning is however not identical. A long period of time can sometimes pass before an owner decides to decommission and dismantle a facility. The conditions will also differ depending on the strategy, 'immediate dismantling' or 'safe enclosure'. If immediate dismantling is chosen the site can reach 'green-field status' in less than ten years. 'Safe enclosure', however, seems to be the most common strategy. There are several pathways, but in general a safe store is constructed, enabling the active parts to remain in safe and waterproof conditions for a longer period of

  15. Introduction: Imams in Western Europe

    DEFF Research Database (Denmark)

    Vinding, Niels Valdemar; Hashas, Mohammed; de Ruiter, Jan Jaap

    2018-01-01

    by this source of religious authority in the context of the secular-liberal societies of Western Europe since the Second World War and the subsequent migration and refugee flows. At the same time, this research also serves to highlight secular-liberal institutions and their adaptation, or lack thereof......, to the multiculturalism that characterizes Western European states. The social facts of globalization, transnational migration, and various interpretations of secularism have challenged the visibility of religion in the public sphere in Western societies. This has most importantly and urgently required religious...

  16. Application of a Reverse Line Blot hybridisation assay for the species-specific identification of cyathostomins (Nematoda, Strongylida) from benzimidazole-treated horses in the Slovak Republic.

    Science.gov (United States)

    Cernanská, Dana; Paoletti, Barbara; Králová-Hromadová, Ivica; Iorio, Raffaella; Cudeková, Patrícia; Milillo, Piermarino; Traversa, Donato

    2009-03-09

    Five horse farms located in eastern Slovakia were investigated for the presence of benzimidazole-resistant strongyles by faecal egg count reduction test and egg hatch assay. Coprocultures were prepared for each farm from faecal samples taken pre- and post-treatment and harvested larvae were molecularly examined with a Reverse Line Blot assay. Faecal egg count reduction values ranged from 0 to 52.5% and all farms were positive for benzimidazole-resistant cyathostomins. Seven benzimidazole-resistant cyathostomin species were molecularly identified on farms before and also after treatment. These data demonstrate that resistance to benzimidazoles is well established in cyathostomin populations from horse farms in the Slovak Republic and that the molecular assay was able to determine the species-specific distribution of resistant cyathostomins under field conditions.

  17. Western Australian food security project

    Directory of Open Access Journals (Sweden)

    Maycock Bruce

    2007-08-01

    Full Text Available Abstract Background The aim of the Western Australian (WA Food Security Project was to conduct a preliminary investigation into issues relating to food security in one region within the Perth metropolitan area in Western Australia. The first phase of the project involved a food audit in one lower income area that was typical of the region, to identify the range, variety and availability of foods in the region. Methods A comprehensive food audit survey was provided to all food outlet owners/operators in one lower socio-economic region within the City of Mandurah (n = 132 outlets. The purpose of the survey was to investigate the range, variety and availability of foods in the Mandurah region as well as examining specific in-store characteristics such as the types of clientele and in-store promotions offered. Surveys were competed for 99 outlets (response rate = 75%. Results The range of foods available were predominantly pre-prepared with more than half of the outlets pre-preparing the majority of their food. Sandwiches and rolls were the most popular items sold in the outlets surveyed (n = 51 outlets followed by pastries such as pies, sausage rolls and pasties (n = 33 outlets. Outlets considered their healthiest food options were sandwiches or rolls (n = 51 outlets, salads (n- = 50 outlets, fruit and vegetables (n = 40 outlets, seafood (n = 27 outlets, meats such as chicken (n = 26 outlets and hot foods such as curries, soups or quiches (n = 23 outlets. The majority of outlets surveyed considered pre-prepared food including sandwiches, rolls and salads, as healthy food options regardless of the content of the filling or dressings used. Few outlets (n = 28% offered a choice of bread type other than white or wholemeal. High fat pastries and dressings were popular client choices (n = 77% as were carbonated drinks (n = 88% and flavoured milks (n = 46%. Conclusion These findings clearly indicate the need for further investigation of the impact of

  18. Western Pacific Typhoon Aircraft Fixes

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — Western Pacific typhoon aircraft reconnaissance data from the years 1946 - 1965 and 1978, excluding 1952, were transcribed from original documents, or copy of...

  19. The Western Sahara conflict I

    African Journals Online (AJOL)

    Polisario unilaterally declared a "Saharawi Arab Democratic Republic" (27 Feb. 1976). Since .... Furthermore, Frente Polisario managed to keep the Western Sahara question on .... these countries and their fragile ethnic and political balance.

  20. Modeling Wildfire Hazard in the Western Hindu Kush-Himalayas

    Science.gov (United States)

    Bylow, D.

    2012-12-01

    Wildfire regimes are a leading driver of global environmental change affecting a diverse array of global ecosystems. Particulates and aerosols produced by wildfires are a primary source of air pollution making the early detection and monitoring of wildfires crucial. The objectives of this study were to model regional wildfire potential and identify environmental, topological, and sociological factors that contribute to the ignition of wildfire events in the Western Hindu Kush-Himalayas of South Asia. The environmental, topological, and sociological factors were used to model regional wildfire potential through multi-criteria evaluation using a method of weighted linear combination. Moderate Resolution Imaging Spectroradiometer (MODIS) and geographic information systems (GIS) data were integrated to analyze regional wildfires and construct the model. Model validation was performed using a holdout cross validation method. The study produced a significant model of wildfire potential in the Western Hindu Kush-Himalayas.; Western Hindu Kush-Himalayas ; Western Hindu Kush-Himalayas Wildfire Potential

  1. Western forests and air pollution

    International Nuclear Information System (INIS)

    Olson, R.K.; Binkley, D.; Boehm, M.

    1992-01-01

    The book addresses the relationships between air pollution in the western United States and trends in the growth and condition of Western coniferous forests. The major atmospheric pollutants to which forest in the region are exposed are sulfur and nitrogen compounds and ozone. The potential effects of atmospheric pollution on these forests include foliar injury, alteration of growth rates and patterns, soil acidification, shifts in species composition, and modification of the effects of natural stresses

  2. Western Hemisphere Knowledge Partnerships

    Science.gov (United States)

    Malone, T. F.

    2001-05-01

    , and application of knowledge concerning the nature of -- and interaction among -- matter, living organisms, energy, information, and human behavior. This strategy calls for innovative partnerships among the physical, biological, health, and social sciences, engineering, and the humanities. New kinds of partnership must also be forged among academia, business and industry, governments, and nongovernmental organizations. Geophysicists can play an important role in these partnerships. A focus for these partnerships is to manage the individual economic productivity that drives both human development and global change. As world population approaches stability during the twenty-first century, individual economic productivity will be the critical link between the human and the natural systems on planet Earth. AGU is among a core group of individuals and institutions proposing Western Hemisphere Knowledge Partnerships (WHKP) to test the hypothesis that knowledge, broadly construed, is an important organizing principle in choosing a path into the future. The WHKP agenda includes: (1) life-long learning, (2) the health and resilience of natural ecosystems, (3) eco-efficiency in economic production and consumption, (4) extension of national income accounts, (5) environmentally benign sources of energy, (6) delivery of health care, (7) intellectual property rights, and (8) networks for action by local communities.Collaboratories and distance education technologies will be major tools. A panel of experts will explore this proposal.

  3. Note sur la présence de lames aménagées par technique de Kostienki dans les couches gravettiennes du Blot (Cerzat,Haute-Loire).

    OpenAIRE

    Klaric , Laurent

    2000-01-01

    International audience; The unprecedented presence of Kostienki-technique prepared blades (also called Kostienki knives) in the Gravettian layers at Le Blot leads to a new analysis of these artefacts. Thorough technological study has pointed to the possible role of these items as cores, in association with or complementary to burin-forms, in particular context of backed-bladelet production. Le Blot is the second French site yielding such artefacts, the other being Corbiac (Dordogne). The aim ...

  4. Porcine Cysticercosis: Possible Cross-Reactivity of Taenia hydatigena to GP50 Antigen in the Enzyme-Linked Immunoelectrotransfer Blot Assay.

    Science.gov (United States)

    Muro, Claudio; Gomez-Puerta, Luis A; Flecker, Robert H; Gamboa, Ricardo; Barreto, Percy Vilchez; Dorny, Pierre; Tsang, Victor C W; Gilman, Robert H; Gonzalez, Armando E; Garcia, Hector H; O'Neal, Seth E; For The Cysticercosis Working Group In Peru

    2017-12-01

    The lentil lectin glycoprotein enzyme-linked immunoelectrotransfer blot (LLGP EITB, reported sensitivity 99% and specificity 100%) is used as a serologic marker of exposure to Taenia solium in pigs. However, only a limited number of parasites have been evaluated for cross reactivity. Pigs may host other related cestode infections, including Taenia hydatigena, which have not been formally evaluated for cross-reactions. We investigated a corral in Tumbes, Peru, a region where a cysticercosis elimination demonstration project was completed in 2012. In this corral, 14/19 (73.7%) 6-8-week-old piglets were reactive to GP50 on LLGP EITB, and all had circulating Taenia sp. antigens. From eight necropsied piglets; four were infected with T. hydatigena metacestodes whereas none had evidence of T. solium infection. Two resident dogs were subsequently confirmed to have T. hydatigena taeniasis. These results suggest GP50 cross-reactivity in T. hydatigena- infected pigs, although controlled experimental infection is needed to confirm this hypothesis.

  5. Seabird Colonies in Western Greenland

    DEFF Research Database (Denmark)

    Boertmann, D.; Mosbech, A.; Falk, K.

    About 1 million seabirds (indvs) breed in 1032 colonies distributed along the coasts of western Greenland (Fig. 1). However, this figure does not include the little auk colonies in Avanersuaq. These colonies are roughly estimated to hold about 20 mill. pairs. All the basic information on seabird...... colonies in Greenland is compiled in a database maintained by NERI-AE. This report presents data on distribution, population numbers and population trends of 19 species of breeding colonial seabirds in western Greenland. Distributions are depicted on maps in Fig. 18-39. It is apparent that the major...... colonies are found in the northern part of the region, viz. Upernavik and Avanersuaq. The numbers of birds recorded in the database for each species are presented in Tab. 4, and on the basis of these figures estimates of the populations in western Greenland are given (Tab. 5). The most numerous species...

  6. The Western Denmark Heart Registry

    DEFF Research Database (Denmark)

    Schmidt, Morten; Maeng, Michael; Madsen, Morten

    2018-01-01

    The WDHR (Western Denmark Heart Registry) is a seminational, multicenter-based registry with longitudinal registration of detailed patient and procedure data since 1999. The registry includes as of January 1, 2017 approximately 240,000 coronary angiographies, 90,000 percutaneous coronary interven......The WDHR (Western Denmark Heart Registry) is a seminational, multicenter-based registry with longitudinal registration of detailed patient and procedure data since 1999. The registry includes as of January 1, 2017 approximately 240,000 coronary angiographies, 90,000 percutaneous coronary...

  7. Western Canada Sedimentary Basin competitiveness

    International Nuclear Information System (INIS)

    Millar, R.H.G.

    1996-01-01

    Recent dramatic expansion of the natural gas industry in the Western Canada Sedimentary Basin provided ample proof of the potential of this area for further development of natural gas supply. However, the inherent competitive advantages provided by the Western Canada Sedimentary Basin were said to have been offset by low netback prices resulting in poor producer economics when competitiveness is measured by availability of opportunities to find and develop gas supply at costs low enough to ensure attractive returns. Technology was identified as one of the key elements in improving basin competitiveness, but the greatest potential lies in reduced transportation costs and increased access to North American market centres. 8 figs

  8. Eastern and western happiness in work behavior

    Directory of Open Access Journals (Sweden)

    Jaroslava Kubátová

    2017-10-01

    Full Text Available The purpose of this article is to clarify what motivates East Asian work behavior. The research question is: How can a Western manager better understand work behavior and motivation in East Asian cultures? Knowledge about national culture, motivation and the concept of happiness are connected via deductive and comparative methods, while pointing out their new connections and relations. We argue that work motivation in East Asian cultures can be explained using the self-concept-based motivation meta-theory as it corresponds to the East Asian concept of face and that the East Asian self-concept originates with the Eastern concept of happiness.

  9. Phosphine reduced IgG. A new method for 99mTc labeling immunoglobulins

    International Nuclear Information System (INIS)

    Arteaga de Murphy, C.; Melendez-Alafort, L.; Martinez-Rivero, O.; Gomez, E.; Ferro-Flores, G.

    1997-01-01

    A new technetium labeling method for immunoglobulins reduced with tris(2-carboxy-ethyl)phosphine hydrochloride is presented. The Sandoglobulina IgG source was assayed for purity and optimum reagent's concentration and incubation times were determined. It was purified by column chromatography and labelled with Sn 2+ reduced technetium in the presence of MDP. The kit is easy to prepare, labeling efficiency is >(97±1.9)% and stable for 6 hours.The immunoreactivity of the 99 Tc-IgG was verified by electrophoresis and Western blot tests. The IgG retained its structure after both the reducing and labeling processes and it was the only labeled species. (author)

  10. Phosphine reduced IgG. A new method for {sup 99m}Tc labeling immunoglobulins

    Energy Technology Data Exchange (ETDEWEB)

    Arteaga de Murphy, C; Melendez-Alafort, L [Radiofarmacia Departamento de Medicina Nuclear, Instituto Nacional de Nutricion Salvador Zubiran, Mexico (Mexico); Martinez-Rivero, O [Laboratorio de Quimica Organica, Facultad de Quimica, Universidad de la Habana, Habana (Cuba); Gomez, E [Departamento de Fisiologia de la Nutricion, Instituto Nacional de Nutricion Salvador Zubiran, Mexico (Mexico); Ferro-Flores, G [Depeartamento del Reactor y Materiales Radioactivos, Instituto Nacional de Investigaciones Nucleares, Mexico (Mexico)

    1997-09-01

    A new technetium labeling method for immunoglobulins reduced with tris(2-carboxy-ethyl)phosphine hydrochloride is presented. The Sandoglobulina IgG source was assayed for purity and optimum reagent`s concentration and incubation times were determined. It was purified by column chromatography and labelled with Sn{sup 2+} reduced technetium in the presence of MDP. The kit is easy to prepare, labeling efficiency is >(97{+-}1.9)% and stable for 6 hours.The immunoreactivity of the {sup 99}Tc-IgG was verified by electrophoresis and Western blot tests. The IgG retained its structure after both the reducing and labeling processes and it was the only labeled species. (author). 11 refs.

  11. Smoke exposure at western wildfires.

    Science.gov (United States)

    Timothy E. Reinhardt; Roger D. Ottmar

    2000-01-01

    Smoke exposure measurements among firefighters at wildfires in the Western United States between 1992 and 1995 showed that altogether most exposures were not significant, between 3 and 5 percent of the shift-average exposures exceeded occupational exposure limits for carbon monoxide and respiratory irritants. Exposure to benzene and total suspended particulate was not...

  12. Shell Trumpets from Western Mexico

    Directory of Open Access Journals (Sweden)

    Robert Novella

    1991-11-01

    Full Text Available Marine shells have been used as musical instruments in almost all parts of the world (Izikowitz 1935, including Mesoamerica, where large univalves, also called conch shells in the literature, had a utilitarian function as trumpets. Their use is well documented in most cultural areas of Mesoamerica, as in Western Mexico, through their various occurrences in archaeological contexts and museums collections.

  13. Outbreak of Sporotrichosis, Western Australia

    Science.gov (United States)

    Feeney, Kynan T.; Whittle, Amanda J.; Altman, Shelley A.; Speers, David J.

    2007-01-01

    A cluster of sporotrichosis cases occurred in the Busselton-Margaret River region of Western Australia from 2000 to 2003. Epidemiologic investigation and mycologic culture for Sporothrix schenckii implicated hay initially distributed through a commercial hay supplier as the source of the outbreak. Declining infection rates have occurred after various community measures were instigated. PMID:17953099

  14. ALIENS IN WESTERN STREAM ECOSYSTEMS

    Science.gov (United States)

    The USEPA's Environmental Monitoring and Assessment Program conducted a five year probability sample of permanent mapped streams in 12 western US states. The study design enables us to determine the extent of selected riparian invasive plants, alien aquatic vertebrates, and some ...

  15. Pigmentary disorders in Western countries

    NARCIS (Netherlands)

    van der Veen, J. P. Wietze

    2007-01-01

    Countries in the so-called "Western" world, especially in Europe, witnessed a dramatic change in ethnic backgrounds of their populations starting in the last decennia of the last century. This had repercussions on various aspects of our society, including medical practice. In dermatology for

  16. Gendering Citizenship in Western Europe

    DEFF Research Database (Denmark)

    Siim, Birte; Lister, Ruth; Williams, Fiona

    The first part of the book clarifies the ways that the concept of citizenship has developed historically and is understood today in a range of Western European welfare states. It elaborates on the contempory framing of debates and struggles around citizenship. This provides a framework for thee...

  17. Western Transitology and Chinese Reality

    DEFF Research Database (Denmark)

    Brødsgaard, Kjeld Erik

    It is the object of considerable debate in Western scholarship whether an authoritarian political order dominated by a strong communist party can continue to exist in China given the many challenges stemming from internal reform and the impact of globalization. Will China eventually turn democratic...

  18. Skywatch: The Western Weather Guide.

    Science.gov (United States)

    Keen, Richard A.

    The western United States is a region of mountains and valleys with the world's largest ocean next door. Its weather is unique. This book discusses how water, wind, and environmental conditions combine to create the climatic conditions of the region. Included are sections describing: fronts; cyclones; precipitation; storms; tornadoes; hurricanes;…

  19. The Shape of a Western

    DEFF Research Database (Denmark)

    Nielsen, Jakob Isak

    2007-01-01

    The article is written for an issue on the Western genre. By means of a comparative analysis of Winchester '73 (Anthony Mann, 1950) and The Man from Laramie (Anthony Mann, 1955) the article accounts for stylistic, narrative, generic and theoretical implications of the transition from Academy ratio...

  20. Accurate and Practical Identification of 20 Fusarium Species by Seven-Locus Sequence Analysis and Reverse Line Blot Hybridization, and an In Vitro Antifungal Susceptibility Study▿†

    Science.gov (United States)

    Wang, He; Xiao, Meng; Kong, Fanrong; Chen, Sharon; Dou, Hong-Tao; Sorrell, Tania; Li, Ruo-Yu; Xu, Ying-Chun

    2011-01-01

    Eleven reference and 25 clinical isolates of Fusarium were subject to multilocus DNA sequence analysis to determine the species and haplotypes of the fusarial isolates from Beijing and Shandong, China. Seven loci were analyzed: the translation elongation factor 1 alpha gene (EF-1α); the nuclear rRNA internal transcribed spacer (ITS), large subunit (LSU), and intergenic spacer (IGS) regions; the second largest subunit of the RNA polymerase gene (RPB2); the calmodulin gene (CAM); and the mitochondrial small subunit (mtSSU) rRNA gene. We also evaluated an IGS-targeted PCR/reverse line blot (RLB) assay for species/haplotype identification of Fusarium. Twenty Fusarium species and seven species complexes were identified. Of 25 clinical isolates (10 species), the Gibberella (Fusarium) fujikuroi species complex was the commonest (40%) and was followed by the Fusarium solani species complex (FSSC) (36%) and the F. incarnatum-F. equiseti species complex (12%). Six FSSC isolates were identified to the species level as FSSC-3+4, and three as FSSC-5. Twenty-nine IGS, 27 EF-1α, 26 RPB2, 24 CAM, 18 ITS, 19 LSU, and 18 mtSSU haplotypes were identified; 29 were unique, and haplotypes for 24 clinical strains were novel. By parsimony informative character analysis, the IGS locus was the most phylogenetically informative, and the rRNA gene regions were the least. Results by RLB were concordant with multilocus sequence analysis for all isolates. Amphotericin B was the most active drug against all species. Voriconazole MICs were high (>8 μg/ml) for 15 (42%) isolates, including FSSC. Analysis of larger numbers of isolates is required to determine the clinical utility of the seven-locus sequence analysis and RLB assay in species classification of fusaria. PMID:21389150

  1. Population models of burrowing mayfly recolonization in Western Lake Erie

    Science.gov (United States)

    Madenjian, C.P.; Schloesser, D.W.; Krieger, K.A.

    1998-01-01

    Burrowing mayflies, Hexagenia spp. (H. limbata and H. rigida), began recolonizing western Lake Erie during the 1990s. Survey data for mayfly nymph densities indicated that the population experienced exponential growth between 1991 and 1997. To predict the time to full recovery of the mayfly population, we fitted logistic models, ranging in carrying capacity from 600 to 2000 nymphs/m2, to these survey data. Based on the fitted logistic curves, we forecast that the mayfly population in western Lake Erie would achieve full recovery between years 1998 and 2000, depending on the carrying capacity of the western basin. Additionally, we estimated the mortality rate of nymphs in western Lake Erie during 1994 and then applied an age-based matrix model to the mayfly population. The results of the matrix population modeling corroborated the exponential growth model application in that both methods yielded an estimate of the population growth rate, r, in excess of 0.8 yr-1. This was the first evidence that mayfly populations are capable of recolonizing large aquatic ecosystems at rates comparable with those observed in much smaller lentic ecosystems. Our model predictions should prove valuable to managers of power plant facilities along the western basin in planning for mayfly emergences and to managers of the yellow perch (Perca flavescens) fishery in western Lake Erie.

  2. The study for the Spatial Distribution Pattern of NDVI in the Western of Jilin Province

    Science.gov (United States)

    Yang, Shu-jie; Li, Xiao-dong; Yan, Shou-gang

    2018-02-01

    Using methods of spatial autocorrelation analysis and trend analysis, the paper studies the spatial distribution pattern of NDVI based on the GIMMS NDVI dataset (1998-2008), in Western Jilin. The maximum value for 15d is got through the method of MAX processing. Results show that: the NDVI in growing season shows a rising trend in western Jilin in 1998-2008. In the study area, the NDVI in Western Jilin shows positive spatial autocorrelation in the whole region, but the partial NDVI is apt to scattered distribution, which means the vegetation cover of Western Jilin is generally fragmental.

  3. A simple electroelution method for rapid protein purification: isolation and antibody production of alpha toxin from Clostridium septicum

    Directory of Open Access Journals (Sweden)

    Lorena Vázquez-Iglesias

    2017-06-01

    Full Text Available Clostridium septicum produces a number of diseases in human and farm animals which, in most of the cases, are fatal without clinical intervention. Alpha toxin is an important agent and the unique lethal virulent factor produced by Clostridium septicum. This toxin is haemolytic, highly lethal and necrotizing activities but is being used as an antigen to develop animal vaccines. The aim of this study was to isolate the alpha toxin of Clostridium septicum and produce highly specific antibodies against it. In this work, we have developed a simple and efficient method for alpha toxin purification, based on electroelution that can be used as a time-saving method for purifying proteins. This technique avoids contamination by other proteins that could appear during other protein purification techniques such chromatography. The highly purified toxin was used to produce polyclonal antibodies. The specificity of the antibodies was tested by western blot and these antibodies can be applied to the quantitative determination of alpha toxin by slot blot.

  4. Iodine-123 in Western Europe

    International Nuclear Information System (INIS)

    Qaim, S.M.; Stoecklin, G.; Weinreich, R.

    1976-08-01

    The major object of this panel was to obtain information on the state of art of Iodine-123 production in Western Europe. Technical, medical and organizational problems were discussed extensively during the one-day meeting and a stimulating exchange of information between the various 123 I-producers and users has been initiated. Some specific examples of medical application were also included in order to get a feeling of the degree of acceptance by the medical community and the demand for this isotope. The meeting clearly demonstrated the great demand for this isotope but it also showed that the present rate of production is well below the demand. In order to fill this gap, not only further technical development is needed but also the organizational question of distribution has to be solved, perhaps within a network of collaborating cyclotrons, a task which is considerably more difficult in Western Europe than in the USA. (orig./HP) [de

  5. The western Veil nebula (Image)

    Science.gov (United States)

    Glenny, M.

    2009-12-01

    The western Veil nebula in Cygnus. 15-part mosaic by Mike Glenny, Gloucestershire, taken over several months mostly in the autumn of 2008. 200mm LX90/f10 autoguided, Meade UHC filter, 0.3xFR/FF, Canon 20Da DSLR. Exposures each typically 10x360 secs at ISO1600, processed in Registax4, PixInsight (for flat field correction) & Photoshop CS.

  6. Ethics, Gesture and the Western

    Directory of Open Access Journals (Sweden)

    Michael Minden

    2017-06-01

    Full Text Available This paper relates the Western Movie to Agamben’s implied gestural zone between intention and act. Film is important in the realisation of this zone because it was the first means of representation to capture the body in movement. The Western movie explores the space of ethical indistinction between the acts of individual fighters and the establishment of a rule of law, or putting this another way, between violence and justice. Two classic examples of an archetypal Western plot (Shane, 1953 and Unforgiven, 1991 that particularly embodies this are cited. In both a gunfighter who has forsworn violence at the start is led by the circumstances of the plot to take it up once more at the conclusion. In these terms all the gestures contained between these beginning- and end-points are analysable as an ethics of gesture because, captured as gestures, they occupy the human space between abstraction and action, suspended between them, and reducible to neither.  David Foster Wallace's definition of this narrative arc in Infinite Jest (and embodied in it is adduced in order to suggest a parallel between Agamben's notion of an ethics of gesture, and an ethics of genre.

  7. Factors affecting the use of prenatal care by non-western women in industrialized western countries: a systematic review

    Science.gov (United States)

    2013-01-01

    Background Despite the potential of prenatal care for addressing many pregnancy complications and concurrent health problems, non-western women in industrialized western countries more often make inadequate use of prenatal care than women from the majority population do. This study aimed to give a systematic review of factors affecting non-western women’s use of prenatal care (both medical care and prenatal classes) in industrialized western countries. Methods Eleven databases (PubMed, Embase, PsycINFO, Cochrane, Sociological Abstracts, Web of Science, Women’s Studies International, MIDIRS, CINAHL, Scopus and the NIVEL catalogue) were searched for relevant peer-reviewed articles from between 1995 and July 2012. Qualitative as well as quantitative studies were included. Quality was assessed using the Mixed Methods Appraisal Tool. Factors identified were classified as impeding or facilitating, and categorized according to a conceptual framework, an elaborated version of Andersen’s healthcare utilization model. Results Sixteen articles provided relevant factors that were all categorized. A number of factors (migration, culture, position in host country, social network, expertise of the care provider and personal treatment and communication) were found to include both facilitating and impeding factors for non-western women’s utilization of prenatal care. The category demographic, genetic and pregnancy characteristics and the category accessibility of care only included impeding factors. Lack of knowledge of the western healthcare system and poor language proficiency were the most frequently reported impeding factors. Provision of information and care in women’s native languages was the most frequently reported facilitating factor. Conclusion The factors found in this review provide specific indications for identifying non-western women who are at risk of not using prenatal care adequately and for developing interventions and appropriate policy aimed at

  8. Annosus Root disease of Western Conifers (FIDL)

    Science.gov (United States)

    Craig L. Schmitt; John R. Parmeter; John T. Kliejunas

    2000-01-01

    Annosus root disease is found on all western conifer species but is of most concern on true firs, hemlocks, and pines. Incense cedar, coast redwood and sequoia are sometimes infected in California. Western juniper is infected throughout its range. Annosus is common and causes extensive decay in old-growth western and mountain hemlock stands. Many mixed conifer stands...

  9. Women and Islam in the Western Media

    NARCIS (Netherlands)

    Vintges, K.; Ennaji, M.

    2016-01-01

    This chapter focuses on recent debates on women and Islam as framed in the Western media. The first section discusses the way these debates are organized through the presentations and self-presentations of Muslim women in the Western media. The second section (titled “The Restyling of Western

  10. Preliminary Strategic Environmental Assessment of the Great Western Development Strategy: Safeguarding Ecological Security for a New Western China

    Science.gov (United States)

    Li, Wei; Liu, Yan-Ju; Yang, Zhifeng

    2012-02-01

    The Great Western Development Strategy (GWDS) is a long term national campaign aimed at boosting development of the western area of China and narrowing the economic gap between the western and the eastern parts of China. The Strategic Environmental Assessment (SEA) procedure was employed to assess the environmental challenges brought about by the western development plans. These plans include five key developmental domains (KDDs): water resource exploitation and use, land utilization, energy generation, tourism development, and ecological restoration and conservation. A combination of methods involving matrix assessment, incorporation of expert judgment and trend analysis was employed to analyze and predict the environmental impacts upon eight selected environmental indicators: water resource availability, soil erosion, soil salinization, forest destruction, land desertification, biological diversity, water quality and air quality. Based on the overall results of the assessment, countermeasures for environmental challenges that emerged were raised as key recommendations to ensure ecological security during the implementation of the GWDS. This paper is intended to introduce a consensus-based process for evaluating the complex, long term pressures on the ecological security of large areas, such as western China, that focuses on the use of combined methods applied at the strategic level.

  11. Risk assessment for yellow fever in western and North-Western provinces of Zambia

    Directory of Open Access Journals (Sweden)

    Olusegun A Babaniyi

    2015-01-01

    Full Text Available Background: North-Western and Western provinces of Zambia were reclassified as low-risk areas for yellow fever (YF. However, the current potential for YF transmission in these areas is unclear. Aims: To determine the current potential risk of YF infection. Setting and Design: A cross sectional study was conducted in North-Western and Western provinces of Zambia. Materials and Methods: Samples were tested for both YF virus-specific IgG and IgM antibodies by the ELISA and YF virus confirmation was done using Plaque Reduction Neutralization Test. The samples were also tested for IgG and IgM antibodies against other flaviviruses. Results: Out of the 3625 respondents who participated in the survey, 46.7% were males and 9.4% were aged less than 5 years. Overall, 58.1% of the participants slept under an impregnated insecticide-treated net and 20.6% reported indoor residual spraying of insecticides. A total of 616 (17.0% samples were presumptive YF positive. The prevalence for YF was 0.3% for long-term infection and 0.2% for recent YF infection. None of the YF confirmed cases had received YF vaccine. Prevalence rates for other flaviviruses were 149 (4.1% for Dengue, 370 (10.2% for West Nile and 217 (6.0% for Zika. Conclusion: There is evidence of past and recent infection of YF in both provinces. Hence, they are at a low risk for YF infection. Yellow fever vaccination should be included in the EPI program in the two provinces and strengthen surveillance with laboratory confirmation.

  12. Motives underlying food consumption in the Western Balkans

    NARCIS (Netherlands)

    Mardon, Julie; Thiel, Elise; Laniau, Martine; Sijtsema, Siet; Zimmermann, Karin; Barjolle, Dominique

    2015-01-01

    Objectives: This study aims to identify subgroups of consumers based on the health motives underlying their food choice in Western Balkan Countries. Methods: The survey (n = 2943) was based on the Food Choice Questionnaire (FCQ) and elicited information on socio-demographic characteristics,

  13. Contraceptive choice of women in Akure, Western Nigeria ...

    African Journals Online (AJOL)

    Study design/Setting/Methods: It was a cross-sectional descriptive study involving 400 family planning clients who attended the State Specialist Hospital, Akure, Western Nigeria between July and December 2012 using ... Majority were in the age bracket of 30-39years (53.1%), many had post secondary education (50%).

  14. Aeromycobiota of Western Desert of Egypt | Ismail | African Journal ...

    African Journals Online (AJOL)

    The prevalence of airborne mycobiota at six different regions of Western desert (5 regions) and Eastern desert (1) of Egypt was determined using the exposed-plate method. A total of 44 genera, 102 species and one variety in addition to some unidentified yeasts and dark sterile mycelia were collected. Of the above, only 5 ...

  15. Body Image and Westernization Trends among Japanese Adolescents

    Science.gov (United States)

    Nielson, Hailey E.; Reel, Justine J.; Galli, Nick A.; Crookston, Benjamin T.; Miyairi, Maya

    2013-01-01

    Background and Aims: The purpose of this project was to examine body dissatisfaction and the degree of acculturation to Western media body ideals among Japanese adolescents. Furthermore, sex differences in body esteem were examined between male and female participants. Methods: Male and female participants (N=158) aged 15 to 18 years in Okinawa,…

  16. MARCC (Matrix-Assisted Reader Chromatin Capture): an antibody-free method to enrich and analyze combinatorial nucleosome modifications

    Science.gov (United States)

    Su, Zhangli

    2016-01-01

    Combinatorial patterns of histone modifications are key indicators of different chromatin states. Most of the current approaches rely on the usage of antibodies to analyze combinatorial histone modifications. Here we detail an antibody-free method named MARCC (Matrix-Assisted Reader Chromatin Capture) to enrich combinatorial histone modifications. The combinatorial patterns are enriched on native nucleosomes extracted from cultured mammalian cells and prepared by micrococcal nuclease digestion. Such enrichment is achieved by recombinant chromatin-interacting protein modules, or so-called reader domains, which can bind in a combinatorial modification-dependent manner. The enriched chromatin can be quantified by western blotting or mass spectrometry for the co-existence of histone modifications, while the associated DNA content can be analyzed by qPCR or next-generation sequencing. Altogether, MARCC provides a reproducible, efficient and customizable solution to enrich and analyze combinatorial histone modifications. PMID:26131849

  17. Biophysical characterization data on Aβ soluble oligomers produced through a method enabling prolonged oligomer stability and biological buffer conditions

    Directory of Open Access Journals (Sweden)

    Amanda C. Crisostomo

    2015-09-01

    Aβ1-40 soluble oligomers are produced that are suitable for biophysical studies requiring sufficient transient stability to exist in their “native” conformation in biological phosphate-saline buffers for extended periods of time. The production involves an initial preparation of highly monomeric Aβ in a phosphate saline buffer that transitions to fibrils and oligomers through time incubation alone, without added detergents or non-aqueous chemicals. This criteria ensures that the only difference between initial monomeric Aβ reactant and subsequent Aβ oligomer products is their degree of peptide assembly. A number of chemical and biophysical methods were used to characterize the monomeric reactants and soluble oligomer and amyloid fibril products, including chemical cross-linking, Western blots, fraction solubility, thioflvain T binding, size exclusion chromatography, transmission electron micrscopy, circular dichroism spectroscopy, and fluorescence resonance energy transfer.

  18. Lime stabilization of fine-grained sediments in western Greenland

    DEFF Research Database (Denmark)

    Jørgensen, Anders Stuhr; Ingeman-Nielsen, Thomas; Skels, Peteris

    2011-01-01

    Thick deposits of fine-grained marine sediments exist in large areas of western Greenland. Many places these sediments are located above sea-level, and now complicate construction projects in urban areas. The mineralogy of the fine-grained sediments is very different from European sediments, mainly...... due to the cold climate, and it is therefore of great interest to study possible methods to improve the stability and strength properties. This project includes laboratory studies of lime stabilization on fine-grained marine sediments from Kangerlussuaq, western Greenland. The results have included...

  19. The potential for western Canada to become a leader in electrically powered land transport

    International Nuclear Information System (INIS)

    Perl, A.; Gilbert, R.

    2010-01-01

    Canada's western provinces are among the most oil-intensive jurisdictions in the world. Per capita consumption in 2007 was 70 percent higher than that of eastern Canada. While western Canada will be buffered as a producer of oil, predicted swings in oil prices may disrupt the fiscal capacity of producer provinces. The price of crude oil in western Canada is the same price established in world markets. This paper examined some of the opportunities available for western Canada to move away from oil consumption by becoming a leader in electric traction. Methods of replacing oil-based transportation with the use of electric vehicles were discussed. Western Canada's capacity to produce electricity from renewable resources was evaluated. Economic opportunities related to the electrification of land transport were also examined. The report concluded by stating that a clear vision and policy framework for supporting new technologies for surface transport across western Canada are needed. 72 refs., 4 figs.

  20. A Stochastic Frontier Analysis of Output Level and Growth in Poland and Western Economies

    NARCIS (Netherlands)

    Osiewalski, J.; Koop, G.; Steel, M.F.J.

    1997-01-01

    This paper uses Bayesian stochastic frontier methods to measure the productivity gap between Poland and Western countries that existed before the beginning of the main Polish economic reform. Using data for 20 Western economies, Poland and Yugoslavia (1980-1990) we estimate a translog stochastic

  1. Cone and seed production of western larch in response to girdling and nitrogen fertilization - an update

    Science.gov (United States)

    Russell T. Graham; Jonalea R. Tonn; Theresa B. Jain

    1995-01-01

    Western larch (Larix occidentalis Nutt.) is a sporadic cone and seed producer. Because the species is such an important component of the -Northern Rocky Mountain forests, methods of increasing seed production are needed. Girdling, fertilizing, and a combination of the two were used on 75-year-old western larch in northern Idaho. Girdling at the base of the live crown...

  2. Factors affecting the use of prenatal care by non-western women in industrialized western countries: a systematic review.

    Science.gov (United States)

    Boerleider, Agatha W; Wiegers, Therese A; Manniën, Judith; Francke, Anneke L; Devillé, Walter L J M

    2013-03-27

    Despite the potential of prenatal care for addressing many pregnancy complications and concurrent health problems, non-western women in industrialized western countries more often make inadequate use of prenatal care than women from the majority population do. This study aimed to give a systematic review of factors affecting non-western women's use of prenatal care (both medical care and prenatal classes) in industrialized western countries. Eleven databases (PubMed, Embase, PsycINFO, Cochrane, Sociological Abstracts, Web of Science, Women's Studies International, MIDIRS, CINAHL, Scopus and the NIVEL catalogue) were searched for relevant peer-reviewed articles from between 1995 and July 2012. Qualitative as well as quantitative studies were included. Quality was assessed using the Mixed Methods Appraisal Tool. Factors identified were classified as impeding or facilitating, and categorized according to a conceptual framework, an elaborated version of Andersen's healthcare utilization model. Sixteen articles provided relevant factors that were all categorized. A number of factors (migration, culture, position in host country, social network, expertise of the care provider and personal treatment and communication) were found to include both facilitating and impeding factors for non-western women's utilization of prenatal care. The category demographic, genetic and pregnancy characteristics and the category accessibility of care only included impeding factors.Lack of knowledge of the western healthcare system and poor language proficiency were the most frequently reported impeding factors. Provision of information and care in women's native languages was the most frequently reported facilitating factor. The factors found in this review provide specific indications for identifying non-western women who are at risk of not using prenatal care adequately and for developing interventions and appropriate policy aimed at improving their prenatal care utilization.

  3. Development of high-throughput and high sensitivity capillary gel electrophoresis platform method for Western, Eastern, and Venezuelan equine encephalitis (WEVEE) virus like particles (VLPs) purity determination and characterization.

    Science.gov (United States)

    Gollapudi, Deepika; Wycuff, Diane L; Schwartz, Richard M; Cooper, Jonathan W; Cheng, K C

    2017-10-01

    In this paper, we describe development of a high-throughput, highly sensitive method based on Lab Chip CGE-SDS platform for purity determination and characterization of virus-like particle (VLP) vaccines. A capillary gel electrophoresis approach requiring about 41 s per sample for analysis and demonstrating sensitivity to protein initial concentrations as low as 20 μg/mL, this method has been used previously to evaluate monoclonal antibodies, but this application for lot release assay of VLPs using this platform is unique. The method was qualified and shown to be accurate for the quantitation of VLP purity. Assay repeatability was confirmed to be less than 2% relative standard deviation of the mean (% RSD) with interday precision less than 2% RSD. The assay can evaluate purified VLPs in a concentration range of 20-249 μg/mL for VEE and 20-250 μg/mL for EEE and WEE VLPs. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  4. Western Areas new U plant

    International Nuclear Information System (INIS)

    Anon.

    1982-01-01

    On March 30, 1982 Western Areas Gold Mining Company Limited's uranium plant was officialy opened. The plant is designed to treat 100 000t/month of uranium bearing ore. The majority of this ore is from the Middle Elsburg series, while the miner part comes from routing upgrated Upper Elsburg products into the uranium plant treatment route. The forward leach concept of gold and uranium extraction is adopted, i.e. the gold is extracted before the uranium. The flow of work, instrumentation, electrical installation and other facilities at the plant are also discussed

  5. Nuclear power in western society

    International Nuclear Information System (INIS)

    Franklin, N.L.

    1977-01-01

    The degree to which problems of public acceptance have contributed to the slowdown in progress of nuclear power in Western European countries and the USA is discussed. Some of the effects on the nuclear power industry, i.e. the electrical utilities, the power station suppliers, and the fuel cycle contractors are described. The problem of the lack of public acceptance is examined by consideration of four areas: the position of the employee working in nuclear installations, opposition from the local community, the question of terrorism and its impact on nuclear policy, and finally, what is felt to constitute the greatest anxiety concerning nuclear power, that of proliferation. (U.K.)

  6. Commuting behavior of western U.S. residents

    Energy Technology Data Exchange (ETDEWEB)

    Caviglia, J. [Tennessee Univ., Knoxville, TN (United States)]|[Oak Ridge National Lab., TN (United States)

    1996-06-01

    Estimation and interpretation of commutes to work has been studied extensively with respect to gender, race, and income. While the literature is extensive in these areas, there has been little research on regional differences between US states and territories. Since data which reports the commute to work is in average minutes, the distance traveled is estimated using estimates of the distance between home and work county centroids. The models differ in estimation of in-county commutes. The first assumes that the commute is equal to the radius of the county and the second estimates the commute as a weighted distance based on place location. Two data sets are compared, US National Guard data and US census data. Goal of this paper is to make conclusions about the commuting behavior of western residents through the use of these estimates, and therefore to provide a estimation method for distance commutes which can be used in further research. It is concluded that the radius method of estimation may be an over estimation, in particular in the western states. Since the non-western states are generally more homogeneously populated, this overestimation is not observed. It is recommended that the place location method be used for similar research, in particular studies dealing with western states. Suggestions are made for further research and recommendations are made for the US Army National Guard in regards to recruiting.

  7. Myxomatosis on the Western Plains of Victoria.

    Science.gov (United States)

    Tighe, F G; Edmonds, J W; Nolan, I F; Shepherd, R C; Gocs, A

    1977-10-01

    Myxomatosis on the Western Plains is an enzootic disease in contrast with the epizootic pattern which is general in eastern Australia. The most unusual aspects are the presence of significant numbers of diseased rabbits throughout the winter and the continuously low percentage of rabbits with antibodies to myxoma virus. Climatic and topographic conditions are unsuited to the production of the high densities of mosquitoes necessary for widespread epizootics. Under these conditions the effects of less efficient methods of myxomatosis transmission are apparent. The unusual epidemiology of myxomatosis has resulted in selection for virulence of the virus similar to that which has occurred under summer epizootic conditions. All field strains are now in the mid range of virulence.

  8. Uranium and radon surveys in western Himalaya

    International Nuclear Information System (INIS)

    Virk, H.S.

    1997-01-01

    The water samples from mountain springs, streams and river systems in the western Himalaya were collected and analysed in the laboratory for uranium and radon contents. It is observed that the Himalayan river system is conspicuous by its high dissolved uranium and radium concentration. The water samples contain from 0.89 ppb to 63.4 ppb of uranium and from 34 Bq/I to 364 Bq/I of radon. The radon emanation in soil is measured by the track-etch method, emanometry and alpha-logger technique. The daily and long-term variation of radon was monitored in some mineralized zones of Himachal Pradesh (HP) state with high uranium content in the soil. The maximum values of radon are recorded in Chhinjra, Rameda, Samurkala and Kasol areas of HP. (author)

  9. Comparative Analysis of Music Recordings from Western and Non-Western traditions by Automatic Tonal Feature Extraction

    Directory of Open Access Journals (Sweden)

    Emilia Gómez

    2008-09-01

    Full Text Available The automatic analysis of large musical corpora by means of computational models overcomes some limitations of manual analysis, and the unavailability of scores for most existing music makes necessary to work with audio recordings. Until now, research on this area has focused on music from the Western tradition. Nevertheless, we might ask if the available methods are suitable when analyzing music from other cultures. We present an empirical approach to the comparative analysis of audio recordings, focusing on tonal features and data mining techniques. Tonal features are related to the pitch class distribution, pitch range and employed scale, gamut and tuning system. We provide our initial but promising results obtained when trying to automatically distinguish music from Western and non- Western traditions; we analyze which descriptors are most relevant and study their distribution over 1500 pieces from different traditions and styles. As a result, some feature distributions differ for Western and non-Western music, and the obtained classification accuracy is higher than 80% for different classification algorithms and an independent test set. These results show that automatic description of audio signals together with data mining techniques provide means to characterize huge music collections from different traditions and complement musicological manual analyses.

  10. Combination of PCR targeting the VD2 of omp1 and reverse line blot analysis for typing of urogenital Chlamydia trachomatis serovars in cervical scrape specimens.

    NARCIS (Netherlands)

    Molano, M; Meijer, C.J.L.M.; Morre, S.A.; Pol, R; Brule, van den AJ

    2004-01-01

    50% contained both serovars D and E. The nested VD2 PCR-RLB developed is a simple, fast, and specific method for the identification of individual urogenital C. trachomatis serovars previously detected by using plasmid PCR. Moreover, it is an appropriate method for studying multiple C. trachomatis

  11. Peritoneal Dialysis in Western Countries.

    Science.gov (United States)

    Struijk, Dirk G

    2015-12-01

    Peritoneal dialysis (PD) for the treatment of end-stage renal failure was introduced in the 1960s. Nowadays it has evolved to an established therapy that is complementary to hemodialysis (HD), representing 11% of all patients treated worldwide with dialysis. Despite good clinical outcomes and similar results in patient survival between PD and HD, the penetration of PD is decreasing in the Western world. First the major events in the history of the development of PD are described. Then important insights into the physiology of peritoneal transport are discussed and linked to the changes in time observed in biopsies of the peritoneal membrane. Furthermore, the developments in peritoneal access, more biocompatible dialysate solutions, automated PD at home, the establishment of parameters for dialysis adequacy and strategies to prevent infectious complications are mentioned. Finally non-medical issues responsible for the declining penetration in the Western world are analyzed. Only after introduction of the concept of continuous ambulatory PD by Moncrief and Popovich has this treatment evolved in time to a renal replacement therapy. Of all structures present in the peritoneal membrane, the capillary endothelium offers the rate-limiting hindrance for solute and water transport for the diffusive and convective transport of solutes and osmosis. The functional and anatomical changes in the peritoneal membrane in time can be monitored by the peritoneal equilibrium test. Peritonitis incidence decreased by introduction of the Y-set and prophylaxis using mupirocin on the exit site. The decrease in the proportion of patients treated with PD in the Western world can be explained by non-medical issues such as inadequate predialysis patient education, physician experience and training, ease of HD initiation, overcapacity of in-center HD, lack of adequate infrastructure for PD treatment, costs and reimbursement issues of the treatment. (1) PD is cheaper than HD and provides a

  12. Statistical estimation of future precipitation in south-western Europe taking into account spatial differentiation methods; Abschaetzung einer zukuenftigen Niederschlagsentwicklung mit statistischen Methoden unter Einbezug raeumlicher Differenzierungsverfahren am Beispiel des suedwesteuropaeischen Raums

    Energy Technology Data Exchange (ETDEWEB)

    Woth, K. [GKSS-Forschungszentrum Geesthacht GmbH (Germany). Inst. fuer Kuestenforschung

    2001-07-01

    This study presents a method to identify homogenous sub-regions and their corresponding large-scale sector in statistical-empirical downscaling models systematically. The analysis was done with a gridded observed sea level pressure field (SLP) of the North Atlantic sector as predictor and time series of precipitation at 82 Stations as dependent variable, distributed nearly homogeneously in Spain and the south of France. The precipitation stations were classified in more homogeneous groups by cluster analysis. For some of the resulting groups the domain of the SLP field with the highest influence on rainfall was also identified. In the next step Canonical Correlation Analysis was applied between the groups suitable for a statistical downscaling and their corresponding SLP sector. The result is that the statistical skill in the validation period is increased compared to the standard downscaling model. Last, this statistical downscaling model are applied to a climate change experiment with ECHAM4-OPYC3 under increasing atmospheric CO{sub 2} concentrations. (orig.)

  13. Changes in rRNA levels during stress invalidates results from mRNA blotting: Fluorescence in situ rRNA hybridization permits renormalization for estimation of cellular mRNA levels

    DEFF Research Database (Denmark)

    Hansen, M.C.; Nielsen, A.K.; Molin, Søren

    2001-01-01

    obtained by these techniques are compared between experiments in which differences in growth rates, strains, or stress treatments occur, the normalization procedure may have a significant impact on the results. In this report we present a solution to the normalization problem in RNA slot blotting...... the relative level of rRNA per cell, and slot blotting to rRNA probes, which estimates the level of rRNA per extracted total RNA, the amount of RNA per cell was calculated in a series of heat shock experiments with the gram-positive bacterium Lactococcus lactis. It was found that the level of rRNA per cell...... decreased to 30% in the course of the heat shock. This lowered ribosome level led to a decrease in the total RNA content, resulting in a gradually increasing overestimation of the mRNA levels throughout the experiment. Using renormalized cellular mRNA levels, the HrcA-mediated regulation of the genes...

  14. Western Canada : changing pricing dynamics

    International Nuclear Information System (INIS)

    Frank, B.

    1998-01-01

    Natural gas supply and demand trends in Western Canada are reviewed in a series of overhead viewgraphs. Production versus pipeline capacity, required gas well completions in the WCSB to meet local demand and fill export pipeline capacity to year 2005, NYMEX and AECO price trends during 1995-2000, and the question of what will happen to prices with additional pipeline capacity to the U.S. Midwest were summarized. The best guess is that Midwest prices will need to be high enough to attract marginal supplies from the Gulf, i.e. prices have be around the Henry Hub + five cents/ mmbtu. The new Canadian pipelines, (Northern Border and Alliance) will lower Midwest prices somewhat, but the impact will be modest. Assuming that additional planned pipeline expansion come on-stream, the pressure to expand east of Chicago will be considerable. tabs., figs

  15. Western states uranium resource survey

    International Nuclear Information System (INIS)

    Tinney, J.F.

    1977-01-01

    ERDA's National Uranium Resource Evaluation (NURE) program was established to provide a comprehensive description of uranium resources in the United States. To carry out this task, ERDA has contracted with various facilities, including universities, private companies, and state agencies, to undertake projects such as airborne radiometric surveys, geological and geochemical studies, and the development of advanced geophysical technology. LLL is one of four ERDA laboratories systematically studying uranium distribution in surface water, groundwater, and lake and stream sediments. We are specifically responsible for surveying seven western states. This past year we have designed and installed facilities for delayed-neutron counting and neutron-activation analysis, completed seven orientation surveys, and analyzed several thousand field samples. Full-scale reconnaissance surveys began last fall

  16. [Migratory circuits in western Mexico].

    Science.gov (United States)

    Durand, J

    1986-11-01

    The author examines patterns of internal and international migration in western Mexico. "Drawing on data from different sources and statistics, the essay demonstrates the importance of both types of migration, the changes in endogenous and exogenous factors which have affected the life and the migratory patterns of the population of this region. The migratory circuit being a flow not only of persons, but of goods and capital as well, the cities, specifically that of Guadalajara, have a strategic importance. They fulfill various functions and have become the backbone of the migratory process: they serve as centers for attracting and 'hosting' internal migrants as well as places of origin for other migrants; jumping-off points for international migrants; and the milieu in which many returning migrants of rural origin settle." (SUMMARY IN ENG AND FRE) excerpt

  17. Radiogeochronologic data on rocks of crystalline massifs of Western Carpathians and their interpretation in radiogeochronologic dating

    International Nuclear Information System (INIS)

    Cambel, B.

    1979-01-01

    The results are summed up and evaluated of the measurement of the age of various crystalline rocks in the Western Carpathians using the method of isotope dating. The obtained results show that the rocks in the Western Carpathians were formed in the period of the Variscian and Alpine orogenesis, ie., 90 to 550 million years ago. A list of problems is given which must be resolved using radiogeochronological methods for age determination of geological processes and for specifying the stratigraphy of crystalline rocks in the Western Carpathians. (Ha)

  18. Seroprevalence of antibodies to Borrelia burgdorferi sensu lato in healthy adults from western Norway: risk factors and methodological aspects.

    Science.gov (United States)

    Hjetland, Reidar; Nilsen, Roy M; Grude, Nils; Ulvestad, Elling

    2014-11-01

    The aim of this study was to assess the seroprevalence of antibodies to Borrelia burgdorferi sensu lato in a healthy adult population from Sogn and Fjordane county in western Norway by different assays. Sera from 1213 blood donors at four different blood banks were analysed in Enzygnost Lyme link VlsE/IgG (IgG), Enzygnost Borreliosis IgM (IgM), and Immunetics C6 Lyme ELISA kit (C6). Sera showing positive or grey-zone reactivities were further examined with Borrelia-EUROLine-RN-AT IgG blot and Borrelia-EUROLine-RN-AT IgM blot. The seroprevalences were 9.6%, 8.2%, 8.4%, 6.4% and 5.7%, respectively. The seroprevalence for IgG was lower in the eastern part of the county and in owners of pet animals. It was higher in men, and increased with age and number of tick bites. C6 and IgG gave comparable results. IgM only was found in 4.5%, more often in women, did not increase with age, and showed no relationship with geography, and 56.4% were positive in IgM blot. In conclusion, antibodies to B. burgdorferi s.l. are common in blood donors in western Norway. The results may be used for evaluation of predictive values of test results in patients, as well as a basis for test algorithms in the laboratory. © 2014 APMIS. Published by John Wiley & Sons Ltd.

  19. Th1/Th2 balance and humoral immune response to potential antigens as early diagnostic method of equine Strongylus nematode infection

    Directory of Open Access Journals (Sweden)

    Faten A. M. Abo-Aziza

    2017-06-01

    Full Text Available Aim: The aim of this study was to investigate the early diagnosis of strongyle infection based on early changes in Th1 and Th2 cytokines beside the diagnostic accuracy values and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE and western blotting profiles using prepared strongyles antigens. Materials and Methods: A total of 73 donkeys had a mean age of 4-32 years old were parasitologically examined for strongyle infection. The early changes in Th1 and Th2 cytokines were determined, and the diagnostic accuracy values and SDS-PAGE and western blotting profiles were performed using prepared strongyles antigens; crude somatic Strongylus vulgaris (CSS, excretory-secretory S. vulgaris (ESS, crude somatic Cyathostomins (CSC, and excretory-secretory Cyathostomins (ESC. Results: The results revealed highest 437.04% and lowest 37.81% immunoglobulin G (IgG in high and low egg shedder groups when using ESC and CSS antigens, respectively. Antibodies index for ESS and CSC were significantly higher in moderate egg shedder group while that for ESS and CSC, ESC was significantly higher in high egg shedder group. Tumor necrosis factor alpha (TNF-α/interleukin-4 (IL-4 balance in S. vulgaris infected donkeys was approximately equal in apparently healthy, low and high egg shedder groups while TNF-α < IL-4 in moderate egg shedder. In Cyathostomins infected animals, TNF-α/IL-4 balance was approximately equal in apparently healthy group while it was low in moderate and high egg shedder groups. The diagnostic accuracy showed that the higher specificity (46.6% and prevalence (95.40% were recorded by CSS and ESC antigens, respectively. However, SDS-PAGE and western blotting profiling proved that the band at molecular weight 25 kDa is exhibited by CSS antigen. Conclusion: Combination of detecting level of TNF-α/IL-4 balance, CSS antigen and IgG concentration is good tool for appropriate diagnosis of such infection. More advancement research must be

  20. Evaluation of the BioPlex 2200 syphilis system as a first-line method of reverse-sequence screening for syphilis diagnosis.

    Science.gov (United States)

    Marangoni, Antonella; Nardini, Paola; Foschi, Claudio; Moroni, Alessandra; D'Antuono, Antonietta; Bacchi Reggiani, Letizia; Cevenini, Roberto

    2013-07-01

    Despite recent technological advances, the diagnosis of syphilis remains a challenging enterprise. Actually, most high-volume laboratories have adopted the "reverse algorithm" due several factors, including the potential to automate testing. Recently, immunoassays processed on random-access systems have been proposed as screening tests. The purpose of this study was to evaluate diagnostic performances of BioPlex 2200 Syphilis IgG and BioPlex 2200 Syphilis IgM, tests based on Multiplex Flow technology, in comparison with the performance of Architect Syphilis TP, a chemiluminescent immunoassay for the detection of IgG and/or IgM anti-Treponema pallidum antibodies. A retrospective study was performed with a panel of 100 blood donor sera, a panel of 350 clinical and laboratory-characterized syphilitic sera, and 170 samples obtained from subjects with potentially interfering conditions. Moreover, 200 unselected samples submitted to the Microbiology Laboratory of St. Orsola Hospital in Bologna for routine screening for syphilis were evaluated. As confirmatory tests, T. pallidum hemagglutination and Western blot assays were used. Considering the IgG Western blot (WB) assay to be the gold standard method, BioPlex 2200 Syphilis IgG specificity was far higher than Architect Syphilis TP specificity (89.7% versus 78.4%, respectively), whereas the sensitivity was 100% for both automated methods. Compared to the IgM WB assay, BioPlex 2200 Syphilis IgM performed with a specificity of 94.9%, whereas the sensitivity was 84.8%. Considering the excellent ease of use and automation, the high sample throughput and its valuable analytical performances, BioPlex Syphilis 2200 IgG could represent a suitable choice for high-volume laboratories. BioPlex Syphilis 2200 IgM could be considered a good addition to IgG testing for uncovering active infections.

  1. Expert Western Classical Music Improvisers' Strategies

    Science.gov (United States)

    Després, Jean-Philippe; Burnard, Pamela; Dubé, Francis; Stévance, Sophie

    2017-01-01

    The growing interest in musical improvisation is exemplified by the body of literatures evidencing the positive impacts of improvisation learning on the musical apprentice's aptitudes and the increasing presence of improvisation in Western classical concert halls and competitions. However, high-level Western classical music improvisers' thinking…

  2. Body image in non-western societies

    NARCIS (Netherlands)

    Edmonds, A.; Cash, T.

    2012-01-01

    This article discusses a range of body modification and conceptions of the body in non-Western societies. It also analyzes difficulties in applying the primarily Western psychological notion of body image to different societies. Body modification is a near human universal, but has many meanings and

  3. Malaria in Sokoto, North Western Nigeria

    African Journals Online (AJOL)

    STORAGESEVER

    2009-12-15

    Dec 15, 2009 ... Malaria prevalence studies had been undertaken in many parts of Nigeria but there is probably no data available from the far North Western region. This research study was undertaken to determine the prevalence, monthly distribution of malaria in Sokoto, North Western Nigeria in order to generate base-.

  4. Management strategies for sustainable western water

    Science.gov (United States)

    Scott Tyler; Sudeep Chandra; Gordon Grant

    2017-01-01

    With the effects of the dramatic western US drought still reverberating through the landscape, researchers gathered in advance of the 20th annual Lake Tahoe Summit to discuss western US water issues in the 21st century. This two-day workshop brought together ~40 researchers from universities and agencies (federal and state) to discuss the prospects that...

  5. Health behaviour among non-Western immigrants with Danish citizenship

    DEFF Research Database (Denmark)

    Hansen, Anne R; Ekholm, Ola; Kjøller, Mette

    2008-01-01

    AIMS: To compare belief in own effort to stay healthy, health behaviour and body mass index (BMI) among non-Western immigrants with Danish citizenship and citizens with Danish background. METHODS: Based on the National Health Interview Survey 2005, logistic regression analyses were used to examine...... differences in belief in own effort to stay healthy, in health behaviour and in BMI between 136 non-Western immigrants with Danish citizenship and 9,901 citizens with Danish background in the age group 25-64 years. RESULTS: Non-Western immigrants had lower odds for reporting that own effort is very important...... to maintain good health (odds ratio (OR) 0.45; 95% confidence interval (CI) 0.32-0.62) and for reporting consuming more alcohol on a weekly basis than recommended by the Danish National Board of Health (OR 0.21, 95% CI 0.09-0.51). The odds were higher for non-Western immigrants for than citizens with Danish...

  6. Future of oil and gas development in the western Amazon

    International Nuclear Information System (INIS)

    Finer, Matt; Babbitt, Bruce; Novoa, Sidney; Ferrarese, Francesco; Pappalardo, Salvatore Eugenio; Marchi, Massimo De; Saucedo, Maria; Kumar, Anjali

    2015-01-01

    The western Amazon is one of the world’s last high-biodiversity wilderness areas, characterized by extraordinary species richness and large tracts of roadless humid tropical forest. It is also home to an active hydrocarbon (oil and gas) sector, characterized by operations in extremely remote areas that require new access routes. Here, we present the first integrated analysis of the hydrocarbon sector and its associated road-building in the western Amazon. Specifically, we document the (a) current panorama, including location and development status of all oil and gas discoveries, of the sector, and (b) current and future scenario of access (i.e. access road versus roadless access) to discoveries. We present an updated 2014 western Amazon hydrocarbon map illustrating that oil and gas blocks now cover 733 414 km 2 , an area much larger than the US state of Texas, and have been expanding since the last assessment in 2008. In terms of access, we documented 11 examples of the access road model and six examples of roadless access across the region. Finally, we documented 35 confirmed and/or suspected untapped hydrocarbon discoveries across the western Amazon. In the Discussion, we argue that if these reserves must be developed, use of the offshore inland model—a method that strategically avoids the construction of access roads—is crucial to minimizing ecological impacts in one of the most globally important conservation regions. (letter)

  7. On the Eastern and Western Cultures as Reflected in Differences in Food Culture

    Institute of Scientific and Technical Information of China (English)

    黄卓; 张海南

    2015-01-01

    When talking about differences between Eastern and Western culture,we should first think of the eating cultural differences.There are many differences in Eastern and Western food cultures,in this paper it will introduce the different food concepts,the different eating goals,the different eating habits,etc.A comparison study of Chinese and Western food culture still makes sense through the analysis of cultural differences between Chinese and Western food,we can understand their own cultural traditions in China and the West.At the same time it is able to carry out improvement and innovation of Chinese culture.Throughout the comparisons,coupled with the differences of the concept of Western food culture,objects,methods,ownership and nature,it studies these differences,identifies areas for mastery of the place,promotes cultural exchange.Thus it enables China to the world,and to make the world know China better.

  8. On the Eastern and Western Cultures as Reflected in Differences in Food Culture

    Institute of Scientific and Technical Information of China (English)

    黄卓; 张海南

    2015-01-01

    When talking about differences between Eastern and Western culture,we should first think of the eating cultural differences.There are many differences in Eastern and Western food cultures,in this paper it will introduce the different food concepts,the different eating goals,the different eating habits,etc. A comparison study of Chinese and Western food culture still makes sense through the analysis of cultural differences between Chinese and Western food,we can understand their own cultural traditions in China and the West.At the same time it is able to carry out improvement and innovation of Chinese culture. Throughout the comparisons,coupled with the differences of the concept of Western food culture,objects,methods,ownership and nature,it studies these differences,identifies areas for mastery of the place,promotes cultural exchange.Thus it enables China to the world,and to make the world know China better.

  9. OPG Western Waste Management Facility

    Energy Technology Data Exchange (ETDEWEB)

    Julian, J. [Ontario Power Generation, Western Waste Management Facility, Tiverton, ON (Canada)

    2011-07-01

    The Ontario Power Generation (OPG) Western Waste Management Facility (WWMF) uses a computer based Supervisory Control and Data Acquisition (SCADA) system to monitor its facility, and control essential equipment. In 2007 the WWMF Low and Intermediate Level Waste (L&ILW) technical support section conducted a review of outstanding corrective maintenance work. Technical support divided all work on a system by system basis. One system under review was the Waste Volume Reduction Building (WVRB) control room SCADA system. Technical support worked with control maintenance staff to assess all outstanding work orders on the SCADA system. The assessment identified several deficiencies in the SCADA system. Technical support developed a corrective action plan for the SCADA system deficiencies, and in February of 2008 developed an engineering change package to correct the observed deficiencies. OPG Nuclear Waste Engineering approved the change package and the WVRB Control Room Upgrades construction project started in January of 2009. The WVRB control room upgrades construction work was completed in February of 2009. This paper provides the following information regarding the WWMF SCADA system and the 2009 WVRB Control Room Upgrades Project: A high-level explanation of SCADA system technology, and the various SCADA system components installed in the WVRB; A description of the state of the WVRB SCADA system during the work order assessment, identifying all deficiencies; A description of the new design package; A description of the construction project; and, A list of lessons learned during construction and commissioning, and a path forward for future upgrades. (author)

  10. 2000 Western Canada activity forecast

    International Nuclear Information System (INIS)

    Kuntz, D.L.

    1999-10-01

    All wells drilled in Western Canada during the first nine months of 1999 are listed and sorted into 12 geographical areas used in the Petroleum Services Association of Canada (PSAC) well cost study. Each area represents wells of common drilling, production and depth characteristics. Area totals for well counts and meters drilled were determined from the sorting process. Previous years' activities are reviewed and various operators and PSAC members contacted to review upcoming programs. In addition, trends and other projections were consulted to develop an estimate of drilling activity for the rest of 1999 as well as a projection of drilling activity for 2000. The historical and projected drilling activities were tabulated and plotted for each area. Average drilling costs for each area were determined, and the total expenditures were calculated for each area by multiplying the the projected meterage by the adjusted drilling costs. All costs were allocated to various services and products utilizing percentages determined in the Well Cost Study. During the sorting process, a list was developed of the major operators in each area, which list is included in the report along with average depths and types of wells drilled by the various operators in each area. The costs included in the report include only drilling and completion operations, starting with the building of the location prior to drilling, and ending with the installation of the wellhead after construction. 5 tabs

  11. Western Canada drilling cycle optimization

    International Nuclear Information System (INIS)

    2003-06-01

    The oil and gas industry in western Canada operates in annual and seasonal cycles with peak activity periods that require a large skilled labour force for short periods of time. This study examines why seismic and drilling activity is greatest during the first quarter of the year instead of being distributed evenly over the year. The objective of the study was to provide recommendations that would help optimize the industry cycle. The study includes an analysis of historical trends that validate the industry first quarter peaking activity. It also includes interviews with 36 industry representatives and provides insight and validation of trends. The final phase of the report includes recommendations that both industry and governments may wish to implement. The study includes financial, operational and environmental considerations. It was shown that natural gas directed drilling activity is strongly correlated with changes in natural gas prices. In the case of oil drilling activity, peak activity responds to oil prices from the prior quarter. In general, drilling and seismic costs are higher in the winter months because of increased demand for equipment and services. In addition winter drilling operations require a diesel fired boiler to generate steam. 36 refs., 2 tabs., 52 figs

  12. Western water and climate change

    Science.gov (United States)

    Dettinger, Michael; Udall, Bradley; Georgakakos, Aris P.

    2015-01-01

    The western United States is a region long defined by water challenges. Climate change adds to those historical challenges, but does not, for the most part, introduce entirely new challenges; rather climate change is likely to stress water supplies and resources already in many cases stretched to, or beyond, natural limits. Projections are for continued and, likely, increased warming trends across the region, with a near certainty of continuing changes in seasonality of snowmelt and streamflows, and a strong potential for attendant increases in evaporative demands. Projections of future precipitation are less conclusive, although likely the northernmost West will see precipitation increases while the southernmost West sees declines. However, most of the region lies in a broad area where some climate models project precipitation increases while others project declines, so that only increases in precipitation uncertainties can be projected with any confidence. Changes in annual and seasonal hydrographs are likely to challenge water managers, users, and attempts to protect or restore environmental flows, even where annual volumes change little. Other impacts from climate change (e.g., floods and water-quality changes) are poorly understood and will likely be location dependent.

  13. Heat pumps in western Switzerland

    International Nuclear Information System (INIS)

    Freymond, A.

    2003-01-01

    The past ten years have seen an extraordinary expansion of heat-pump market figures in the western (French speaking) part of Switzerland. Today, more than 14,000 units are in operation. This corresponds to about 18% of all the machines installed in the whole country, compared to only 10 to 12% ten years ago. This success illustrates the considerable know-how accumulated by the leading trade and industry during these years. It is also due to the promotional program 'Energy 2000' of the Swiss Federal Department of Energy that included the heat pump as a renewable energy source. Already in 1986, the Swiss Federal Institute of Technology in Lausanne was equipped with a huge heat pump system comprising two electrically driven heat pumps of 3.5 MW thermal power each. The heat source is water drawn from the lake of Geneva at a depth of 70 meters. An annual coefficient of performance of 4.5 has been obtained since the commissioning of the plant. However, most heat pump installations are located in single-family dwellings. The preferred heat source is geothermal heat, using borehole heat exchangers and an intermediate heat transfer fluid. The average coefficient of performance of these installations has been increased from 2.5 in 1995 to 3.1 in 2002

  14. Risk perception in western Europe

    International Nuclear Information System (INIS)

    Sjoeberg, Lennart

    1999-01-01

    This paper describes empirical work on risk perception and some related dimensions, in particular with regard to radiation and nuclear power hazards. Most of the data cited come from a current CEC project in which 5 countries in Western Europe have participated. Models of risk perception are discussed and some generally valid findings concerning risk perception are summarized. Risk is seen to be a primary factor in many policy matters and clearly, to the public, more important than utility considerations. Previously formulated models (the Psychometric Model and Cultural Theory) are found to be deficient and a much more efficient alternative is suggested. It is stressed that risk perception is of interest foremost because it can be of value to decision makers in making difficult policy decisions in matters of risk. Hence, it is important to ask what facets of perceived risk are most strongly related to demand for risk mitigation. It is found that expected severity of consequences of an hazard is the clearly most important dimension. The paper concludes with a brief summary of a case study of Swedish experience with high-level nuclear waste repository siting

  15. OPG Western Waste Management Facility

    International Nuclear Information System (INIS)

    Julian, J.

    2011-01-01

    The Ontario Power Generation (OPG) Western Waste Management Facility (WWMF) uses a computer based Supervisory Control and Data Acquisition (SCADA) system to monitor its facility, and control essential equipment. In 2007 the WWMF Low and Intermediate Level Waste (L&ILW) technical support section conducted a review of outstanding corrective maintenance work. Technical support divided all work on a system by system basis. One system under review was the Waste Volume Reduction Building (WVRB) control room SCADA system. Technical support worked with control maintenance staff to assess all outstanding work orders on the SCADA system. The assessment identified several deficiencies in the SCADA system. Technical support developed a corrective action plan for the SCADA system deficiencies, and in February of 2008 developed an engineering change package to correct the observed deficiencies. OPG Nuclear Waste Engineering approved the change package and the WVRB Control Room Upgrades construction project started in January of 2009. The WVRB control room upgrades construction work was completed in February of 2009. This paper provides the following information regarding the WWMF SCADA system and the 2009 WVRB Control Room Upgrades Project: A high-level explanation of SCADA system technology, and the various SCADA system components installed in the WVRB; A description of the state of the WVRB SCADA system during the work order assessment, identifying all deficiencies; A description of the new design package; A description of the construction project; and, A list of lessons learned during construction and commissioning, and a path forward for future upgrades. (author)

  16. Data report: western United States

    International Nuclear Information System (INIS)

    Cook, J.R.; Fay, W.M.

    1982-04-01

    This abbreviated summary data report, presents results of ground water and stream surface sediment reconnaissance in the western United States. Surface sediment samples were collected at 67,741 sites, at a target sampling density of one site per 13 square kilometers. Ground water samples were collected at 13,979 sites, and surface water samples were collected at 2,958 sites. Neutron activaton analysis results are given for uranium and 16 other elements in sediments, and for uranium and 9 other elements in waters. Mass spectrometry results are given for helium in ground waters. Supplemental analyses of the sediments for extractable uranium and 22 other elements are given where they are available. Supplemental analyses of water samples for 33 additional elements are also reported where they are available. Analytical data and field measurements are presented in tables on microfiche. Data from ground water sites (on microfiche in pocket) include (1) water chemistry measurements (pH, conductivity, and alkalinity), (2) physical measurements where applicable (water temperature, well description, and scintillometer reading), and (3) elemental analyses (U, Al, Br Cl, Dy, F, Mg, Mn, Na, and V by neutron activation and Ag, Al, As, B, Ba, Be, Ca, Ce, Co, Cr, Cu, Fe, K, Li, Mg, Mn, Mo, Na, Nb, Ni, P, Sc, Se, Si, Sr, Th, Ti, V, Y, Zn, and Zr by spectrophotometry). Helium analyses are given for ground water

  17. Assess the toxycity and hazard class moldboard rocks Western Donbass

    Directory of Open Access Journals (Sweden)

    Kroyik H.A. О. М. , Т. D.

    2011-11-01

    Full Text Available Researches of the contents of water-soluble forms of heavy metals in different types of dumping mine rocks of the Western Donbass are executed. According to existing state sanitary rules and norms two methods calculate indexes of toxicity for lead, cadmium, zinc, cobalt, nickel, copper. The estimation of an opportunity of application of existing norms and rules at definition of a class of danger of firm waste of the coal-mining industry. ,,

  18. Early and Forced Child Marriages in Rural Western Nepal

    OpenAIRE

    Pitambar Acharya; Benjamin Welsh

    2017-01-01

    After reviewing the state of early and forced child marriage (ECM) globally and nationally within Nepal, this research assessed the determinants, consequences and preventive measures of ECM in rural municipalities in Nepal today. This mixed method surveyed 167 households taking 15 % sample from the clusters of three wards of Badhaiyatal Rural Municipality in Bardiya and Dullu Municipality in Dailekh of Western Nepal. Besides household survey, six Focus Group Discussions (FGDs), 16 Key Informa...

  19. Attitudes towards sex selection in the Western world.

    Science.gov (United States)

    Van Balen, Frank

    2006-07-01

    It appears that in most Western countries, son preference is somewhat stronger than daughter preference. However, when one considers the preference of women it looks as though the opposite pattern is emerging. There is a considerable social acceptance of 'light' methods of sex selection (such as diets), even though these methods are not proven to be effective. The inclination to use sperm separation methods appears to be greater in the United States than in some European countries. There are indications that a preference for boys or for girls is associated with attitudes towards technology, child-rearing style and the stereotyping of boys or girls. Copyright 2006 John Wiley & Sons, Ltd.

  20. Fiscal 1998 geothermal development promotion survey. Annex to on-the-ground survey report, geothermal development promotion survey (Part 2. Electromagnetic surveillance using simplified MT method - No. 30: Western part of Hakkoda area); 1989 nendo chinetsu kaihatsu sokushin chosa. Chinetsu kaihatsu sokushin chosa chijo chosa hokokusho futai shiryo (Denji tansa (MT kan'i ho) hokokusho - No.30 Hakkoda seibu chiiki - 2)

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1991-01-01

    An electromagnetic surveillance using the simplified MT (magnetotelluric) method was conducted to help clarify the geothermal structure in the western part of the Hakkoda area, Aomori Prefecture. The survey covered approximately 60 km{sup 2,} and involved 38 survey points, 3 magnetic field components, 2 electric field components, not less than 10 measuring frequencies in the range of 0.01-125 Hz, and a measuring time of not less than 2 hours. An analysis was performed, with data previously collected at 60 points added to the data collected at the said 38 points. As the result, the resistivity structure in this area was divided into 3 layers of high-low-high as described from the surface to the depth, and the middle layer was further divided into 3 layers of high-middle-low and the bottom layer into 2 layers of high-low. The result conformed though roughly to the result obtained by the electrical logging carried out in the structure boring. In the geothermal zone including the Sukayu hot spa in the southeastern part of this area and along the Jogakura valley, there exist a complicated resistivity structure attributable to geothermal activities and a number of discontinuous lines of resistivity running in the directions of WNW-ESE and N-S. This enabled an inference that there is a deep-seated geothermal source between Odake and Akamizusawa extending eastward from the said geothermal zone. (NEDO)

  1. Fiscal 1998 geothermal development promotion survey. Annex to on-the-ground survey report, geothermal development promotion survey (Part 2. Electromagnetic surveillance using simplified MT method - No. 30: Western part of Hakkoda area); 1989 nendo chinetsu kaihatsu sokushin chosa. Chinetsu kaihatsu sokushin chosa chijo chosa hokokusho futai shiryo (Denji tansa (MT kan'i ho) hokokusho - No.30 Hakkoda seibu chiiki - 2)

    Energy Technology Data Exchange (ETDEWEB)

    NONE

    1991-01-01

    An electromagnetic surveillance using the simplified MT (magnetotelluric) method was conducted to help clarify the geothermal structure in the western part of the Hakkoda area, Aomori Prefecture. The survey covered approximately 60 km{sup 2,} and involved 38 survey points, 3 magnetic field components, 2 electric field components, not less than 10 measuring frequencies in the range of 0.01-125 Hz, and a measuring time of not less than 2 hours. An analysis was performed, with data previously collected at 60 points added to the data collected at the said 38 points. As the result, the resistivity structure in this area was divided into 3 layers of high-low-high as described from the surface to the depth, and the middle layer was further divided into 3 layers of high-middle-low and the bottom layer into 2 layers of high-low. The result conformed though roughly to the result obtained by the electrical logging carried out in the structure boring. In the geothermal zone including the Sukayu hot spa in the southeastern part of this area and along the Jogakura valley, there exist a complicated resistivity structure attributable to geothermal activities and a number of discontinuous lines of resistivity running in the directions of WNW-ESE and N-S. This enabled an inference that there is a deep-seated geothermal source between Odake and Akamizusawa extending eastward from the said geothermal zone. (NEDO)

  2. [The beginning of western medical education].

    Science.gov (United States)

    Kee, C D

    1992-01-01

    Our country had quite an advanced system of medical education during the era of the Koryo Kingdom, and during the Choson Dynasty, the Kyong Guk Dae Jon, in which a systematized medical education was clearly described, was compiled in the era of King Sejong. However, the educational system was not for Western medicine. Western medicine was first introduced to our country in the 9th year of King Injo (1631) when Chong Du Won, Yi Yong Jun, etc. returned from Yon Gyong (Beiuin) with Chik Bang Oe Gi. Knowledge of Western medicine was disseminated by Shil Hak (practical learning) scholars who read a translation in Chinese characters, of Chik Bang Oe Gi. Yi Ik (Song Ho), Yi Gyu Gyong (O ju), Choe Han Gi (Hye Gang), Chong Yak Yong (Ta San), etc., read books of Western medicine and introduced in writing the excellent theory of Western medicine. In addition, Yu Hyong Won (Pan Gye), Pak Ji Won (Yon Am), Pak Je Ga (Cho Jong), etc., showed much interest in Western medicine, but no writings by them about western medicine can be found. With the establishment of a treaty of amity with Japan in the 13th year of King Kojong (1876), followed by the succession of amity treaties with Western powers, foreigners including medical doctors were permitted to flow into this country. At that time, doctors Horace N. Allen, W. B. Scranton, John W. Heron, Rosetta Sherwood (Rosetta S. Hall), etc., came to Korea and inaugurated hospitals, where they taught Western medicine to Korean students. Dr. Horace N. Allen, with the permission of king Kojong, established Che Jung Won in April 1885, and in March 1886, he began at the hospital to provide education of Western medicine to Korean students who were recrutied by the Korean Government. However, the education was not conduted on a regular basis, only training them for work as assistants. This is considered to be the pioneer case of Western medical education in this country. Before that time, Japanese medical doctors came to Korea, but there are no

  3. Reproductive health issues in rural Western Kenya

    Directory of Open Access Journals (Sweden)

    Ouma Peter

    2008-03-01

    Full Text Available Abstract Background We describe reproductive health issues among pregnant women in a rural area of Kenya with a high coverage of insecticide treated nets (ITNs and high prevalence of HIV (15%. Methods We conducted a community-based cross-sectional survey among rural pregnant women in western Kenya. A medical, obstetric and reproductive history was obtained. Blood was obtained for a malaria smear and haemoglobin level, and stool was examined for geohelminths. Height and weight were measured. Results Of 673 participants, 87% were multigravidae and 50% were in their third trimester; 41% had started antenatal clinic visits at the time of interview and 69% reported ITN-use. Malaria parasitemia and anaemia (haemoglobin Conclusion In this rural area with a high HIV prevalence, the reported use of condoms before pregnancy was extremely low. Pregnancy health was not optimal with a high prevalence of malaria, geohelminth infections, anaemia and underweight. Chances of losing a child after birth were high. Multiple interventions are needed to improve reproductive health in this area.

  4. The Western Barents Sea: where is the Caledonian Deformation Front?

    Science.gov (United States)

    Shulgin, Alexey; Aarseth, Iselin; Faleide, Jan Inge; Mjelde, Rolf; Huismans, Ritske

    2017-04-01

    The basement architecture below the Paleozoic sedimentary basins is still not fully understood in the Western Barents Sea region. It has been proposed that the early Devonian Caledonian orogeny has formed structural framework over which major basins have developed lately. However, the geometry of the Caledonian suture zone (its location, orientation and the extent of the deformation front) is still poorly constrained and is ambiguous in the Barents Sea. The crustal evolution of the Barents Sea and the basin-basement interaction is heavily dependent on the spatial extent and orientation of the Caledonian Deformation Front (CDF). In 2014 an active marine seismic experiment was conducted in the Western Barents Sea. One of the goals of the experiment is to discriminate between two existing models for orientations of the CDF: north-south from the potential fields data, and southwest-northeast from seismic data. We also aim to constrain the location of the CDF offshore northern Norway. We present the joint interpretation of collocated newly collected wide-angle seismic data (Ocean Bottom Seismometers) and reprocessing of the reflection seismic dataset (Multi-channel seismics) collected in the mid 1980's, using modern computational techniques. The two seismic methods provide best resolution at different depth ranges, and in our modeling we combine the results from the two methods to constrain the location of the CDF along transect running Northwest-Southeast across the Western Barents Sea.

  5. Development of diagnostic RI test method for glutamate decarboxylase (GAD) antibody, an autoantibody of nerve intractable diseases and I-type diabetes

    International Nuclear Information System (INIS)

    Ota, Mitsuhiro; Ota, Kiyoe; Nishimura, Masataka; Ma Jie; Obayashi, Hiroshi; Saita, Takahiko

    1999-01-01

    Characterization of brain GAD derived from various animals was made using anti-GAD65 peptide and anti-GAD67 peptide antibodies, and the effects of the peptide antibodies on GAD activities were investigated. Enzyme fractions of GAD were prepared from the brains of mouse, rat, bovine and humans to perform Western blot analysis and GAD enzyme assay. When the brain homogenate was applied to Western blotting analysis, anti-GAD65 N-peptide antibody and anti-GAD67 N-peptide one specifically reacted with 67 kDa and 65 kDa isoform, respectively, whereas their C-peptide antibodies were reactive to both respective isoforms. There was no difference in each isoform molecular weight among the species. The immuno-specificity of these antipeptide antibodies was confirmed by immune absorbance assay in the presence of each peptide. Then, effects of the anti-peptide antibody on GAD activity were investigated. The activity of GAD immobilized on the column was dose-dependently increased by adding the anti-serum containing GAD65 or GAD67 N-peptide antibody, but the GAD activity was fully inactivated in the presence of GAD67 C-peptide antibody as well as in the normal serum. These results showed that GAD65 and GAD67 could be isolated by selective use of the respective N-peptide antibodies. However, the yield of isolation by antibody affinity column chromatography was considerably low (only several %) and the enzyme activity obtained was almost inactivated. Therefore, further improvement of the isolation method was thought necessary to use for convenient screening. (M.N.)

  6. Development of diagnostic RI test method for glutamate decarboxylase (GAD) antibody, an autoantibody of nerve intractable diseases and I-type diabetes

    Energy Technology Data Exchange (ETDEWEB)

    Ota, Mitsuhiro; Ota, Kiyoe; Nishimura, Masataka; Ma Jie; Obayashi, Hiroshi; Saita, Takahiko [Utano National Hospital, Kyoto (Japan)

    1999-02-01

    Characterization of brain GAD derived from various animals was made using anti-GAD65 peptide and anti-GAD67 peptide antibodies, and the effects of the peptide antibodies on GAD activities were investigated. Enzyme fractions of GAD were prepared from the brains of mouse, rat, bovine and humans to perform Western blot analysis and GAD enzyme assay. When the brain homogenate was applied to Western blotting analysis, anti-GAD65 N-peptide antibody and anti-GAD67 N-peptide one specifically reacted with 67 kDa and 65 kDa isoform, respectively, whereas their C-peptide antibodies were reactive to both respective isoforms. There was no difference in each isoform molecular weight among the species. The immuno-specificity of these antipeptide antibodies was confirmed by immune absorbance assay in the presence of each peptide. Then, effects of the anti-peptide antibody on GAD activity were investigated. The activity of GAD immobilized on the column was dose-dependently increased by adding the anti-serum containing GAD65 or GAD67 N-peptide antibody, but the GAD activity was fully inactivated in the presence of GAD67 C-peptide antibody as well as in the normal serum. These results showed that GAD65 and GAD67 could be isolated by selective use of the respective N-peptide antibodies. However, the yield of isolation by antibody affinity column chromatography was considerably low (only several %) and the enzyme activity obtained was almost inactivated. Therefore, further improvement of the isolation method was thought necessary to use for convenient screening. (M.N.)

  7. Evaluation of Rural Primary Health Care in Western China: A Cross-Sectional Study.

    Science.gov (United States)

    Wang, Manli; Fang, Haiqing; Bishwajit, Ghose; Xiang, Yuanxi; Fu, Hang; Feng, Zhanchun

    2015-10-29

    China's Ministry of Health has enacted Rural Primary Health Care Program (2001-2010) (HCP) guidelines to improve the quality of people's health. However, the program's success in Western China remains unevaluated. Thus, this study aims to begin to fill that gap by analyzing the provision and utilization of Rural Primary Health Care (RPHC) in Western China. A cross-sectional study was conducted to collect secondary data on the socio-economic characteristics, system construction, services use and implementation of RPHC, and the residents' health status of the sampled areas. Four hundred counties from 31 provinces in China were selected via stratified random sampling, including 171 counties from 12 Western provinces. Twenty-seven analysis indicators, covering system construction, services use and implementation of RPHC were chosen to assess Western China's primary health quality. Analysis of Variance (ANOVA) and Least Significant Difference (LSD) methods were used to measure the RPHC disparities between Western and Eastern and Central China. Technique for Order Preference by Similarity to Ideal Solution (TOPSIS) was used to rank Western, Eastern and Central internal provinces regarding quality of their RPHC. Of the 27 indicators, 13 (48.15%) were below the standard in Western China. These focused on rural health service system construction, Chinese medicine services, and public health. In the comparison between Western, Central and Eastern China, 12 indicators had statistical significance (p China, all indicators were statistically significant (p China overall. Western China's RPHC has proceeded well, but remains weaker than that of Eastern and Central China. Differences within Western internal provinces threaten the successful implementation of RPHC.

  8. (via/vili) in khwisero, western kenya

    African Journals Online (AJOL)

    2013-10-10

    Oct 10, 2013 ... KHWISERO, WESTERN KENYA: LESSON FROM THE FIELD AFFECTING POLICY AND PRACTICE. S. K. Ngichabe ... In SSA, cervical cancer affects mostly women in .... inaccessible to the lower socio-economic population.

  9. Ozone climatology over western Mediterranean Sea

    International Nuclear Information System (INIS)

    Pibiri, G.; Randaccio, P.; Serra, A.; Sollai, A.

    1984-01-01

    A preliminary climatology of atmospheric ozone over Western Mediterranean Sea is given by analysis of the upper observations of O 3 carried out at Cagliari-Elmas station from 1968 to 1976. Some peculiarities are here illustrated and discussed

  10. Western Indian Ocean Journal of Marine Science

    African Journals Online (AJOL)

    Western Indian Ocean Journal of Marine Science. Journal Home · ABOUT THIS JOURNAL · Advanced Search · Current Issue · Archives · Journal Home > Vol 3, No 2 (2004) >. Log in or Register to get access to full text downloads.

  11. Western Indian Ocean Journal of Marine Science

    African Journals Online (AJOL)

    Western Indian Ocean Journal of Marine Science. Journal Home · ABOUT THIS JOURNAL · Advanced Search · Current Issue · Archives · Journal Home > Vol 6, No 2 (2008) >. Log in or Register to get access to full text downloads.

  12. Western Alaska ESI: FISHL (Fish Lines)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This data set contains sensitive biological resource data for anadromous fish species in Western Alaska. Vector lines in this data set represent species occurrences...

  13. Western Alaska ESI: LAKES (Lake Polygons)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This data set contains vector polygons representing lakes and land masses used in the creation of the Environmental Sensitivity Index (ESI) for Western Alaska. The...

  14. Western Alaska ESI: FISH (Fish Polygons)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — This data set contains sensitive biological resource data for marine, estuarine, and anadromous fish species in Western Alaska. Vector polygons in this data set...

  15. 108 WESTERN INFLUENCE ON CHINESE AND NIGERIAN ...

    African Journals Online (AJOL)

    Ike Odimegwu

    extinction of indigenous languages and cultures of many societies in ... Ogirisi: a new journal of African studies vol 9 2012. 109 ... Odinye & Odinye: Western Influence on Chinese & Nigerian Cultures. 110 .... The Ibo People and the Europeans.

  16. Extreme winds in the Western North Pacific

    Energy Technology Data Exchange (ETDEWEB)

    Ott, S.

    2006-11-15

    A statistical model for extreme winds in the western North Pacific is developed, the region on the Planet where tropical cyclones are most common. The model is based on best track data derived mostly from satellite images of tropical cyclones. The methods used to estimate surface wind speeds from satellite images is discussed with emphasis on the empirical basis, which, unfortunately, is not very strong. This is stressed by the fact that Japanese and US agencies arrive at markedly different estimates. On the other hand, best track data records cover a long period of time and if not perfect they are at least coherent over time in their imperfections. Applying the the Holland model to the best track data, wind profiles can be assigned along the tracks. From this annual wind speed maxima at any particular point in the region can be derived. The annual maxima, in turn, are fitted to a Gumbel distribution using a generalization Abild's method that allows for data wind collected from multiple positions. The choice of this method is justified by a Monte Carlo simulation comparing it to two other methods. The principle output is a map showing fifty year winds in the region. The method is tested against observed winds from Philippine synoptic stations and fair agreement is found for observed and predicted 48 year maxima. However, the almost biasfree performance of the model could be fortuitous, since precise definitions of 'windspeed' in terms averaging time, height above ground and assumed surface roughness are not available, neither for best tracks nor for the synoptic data. The work has been carried out under Danish Research Agency grant 2104-04-0005 'Offshore wind power' and it also covers the findings and analysis carried out in connection with task 1.6 of the project 'Feasibility Assessment and Capacity Building for Wind Energy Development in Cambodia, The Philippines and Vietnam' during 2005-06 under contract 125-2004 with EU

  17. Niimina Ahubiya: Western Mono Song Genres

    OpenAIRE

    Loether, Christopher

    1993-01-01

    Although Native American communities may lose their ancestral language or other aspects of their traditional culture, music seems to be more resistant to the continual onslaught of the dominant Euro-American culture. Even today, traditional music remains a vital part of Native American communities throughout the United States. In this article I examine one aspect of the musical traditions of the Western Mono, specifically the different types of songs, and their functions within Western Mono s...

  18. Western Radicalization: Rethinking the Psychology of Terrorism

    Science.gov (United States)

    2017-03-01

    to do so, they assert, locks both the researcher and the research subject into a “spiral of ignorance and suspicion.” Moreover, “social identity ...percentage of firefighters who commit this type of arson do so to establish an identity and sense of belonging. Similarly, disconnected Westerners...percentage of firefighters who commit this type of arson do so to establish an identity and sense of belonging. Similarly, disconnected Westerners radicalize

  19. Asian and western Intellectual Capital in encounter

    OpenAIRE

    Boom, van den, Marien; Andriessen, Daan

    2007-01-01

    The purpose of the paper is to start a dialogue about differences between Western and Eastern cultures in the way they conceptualize knowledge and discuss the implications of these differences for a global intellectual capital (IC) theory and practice. A systematic metaphor analysis of the concept of knowledge and IC is used to identify common Western conceptualizations of knowledge in IC literature. A review of philosophical and religious literature was done to identify knowledge conceptuali...

  20. Expert Western Classical Music Improvisers' Strategies

    OpenAIRE

    Despres, JP; Burnard, Pamela Anne; Dube, F; Stevance, S

    2017-01-01

    The growing interest in musical improvisation is exemplified by the body of literatures evidencing the positive impacts of improvisation learning on the musical apprentice’s aptitudes and the increasing presence of improvisation in Western classical concert halls and competitions. However, high-level Western classical music improvisers’ thinking processes are not yet thoroughly documented. As a result of this gap, our research addresses the following question: What strategies are implement...

  1. Bioethical concerns are global, bioethics is Western

    OpenAIRE

    Chattopadhyay, Subrata; De Vries, Raymond

    2008-01-01

    Modern bioethics was born in the West and thus reflects, not surprisingly, the traditions of Western moral philosophy and political and social theory. When the work of bioethics was confined to the West, this background of socio-political theory and moral tradition posed few problems, but as bioethics has moved into other cultures - inside and outside of the Western world - it has become an agent of moral imperialism. We describe the moral imperialism of bioethics, discuss its dangers, and su...

  2. Audiomagnetotelluric investigation of Snake Valley, eastern Nevada and western Utah

    Science.gov (United States)

    McPhee, Darcy K.; Pari, Keith; Baird, Frank

    2009-01-01

    Audiomagnetotelluric (AMT) data along four profiles in western Snake Valley and the corresponding two-dimensional (2-D) inverse models reveal subsurface structures that may be significant to ground-water investigations in the area. The AMT method is a valuable tool for estimating the electrical resistivity of the earth over depth ranges from a few meters to less than one kilometer. The method has the potential to identify faults and stratigraphy within basins of eastern Nevada, thereby helping define the hydrogeologic framework of the region.

  3. Matrix-assisted laser desorption-ionization mass spectrometry peptide mass fingerprinting for proteome analysis: identification efficiency after on-blot or in-gel digestion with and without desalting procedures.

    Science.gov (United States)

    Lamer, S; Jungblut, P R

    2001-03-10

    In theory, peptide mass fingerprinting by matrix assisted laser desorption-ionization mass spectrometry (MALDI-MS) has the potential to identify all of the proteins detected by silver staining on gels. In practice, if the genome of the organism investigated is completely sequenced, using current techniques, all proteins stained by Coomassie Brilliant Blue can be identified. This loss of identification sensitivity of ten to hundred-fold is caused by loss of peptides by surface contacts. Therefore, we performed digestion and transfer of peptides in the lower microl range and reduced the number of steps. The peptide mix obtained from in-gel or on-blot digestion was analyzed directly after digestion or after concentration on POROS R2 beads. Eight protein spots of a 2-DE gel from Mycobacterium bovis BCG were identified using these four preparation procedures for MALDI-MS. Overall, on-blot digestion was as effective as in-gel digestion. Whereas higher signal intensities resulted after concentration, hydrophilic peptides are better detected by direct measurement of the peptide mix without POROS R2 concentration.

  4. Communication and relationship satisfaction in Chinese, Western, and intercultural Chinese-Western couples.

    Science.gov (United States)

    Hiew, Danika N; Halford, W Kim; van de Vijver, Fons J R; Liu, Shuang

    2016-03-01

    The current study compared Chinese, Western, and intercultural Chinese-Western couples' communication and examined how culture moderates the association of communication with relationship satisfaction. We coded the communication of 33 Western couples, 36 Chinese couples, and 54 intercultural Chinese-Western couples when discussing a relationship problem and when reminiscing about positive relationship events. Couples with Chinese female partners showed fewer positive behaviors and more negative behaviors (as classified in existing Western coding systems) than couples with Western female partners. The male partner's culture had few associations with couples' rates of communication behavior. Relationship satisfaction was associated with low rates of negative behaviors and high rates of most of the positive behaviors across cultural groups, and these associations were more evident in problem discussions than positive reminiscences. (c) 2016 APA, all rights reserved).

  5. Exploring Risk Perception and Attitudes to Miscarriage and Congenital Anomaly in Rural Western Kenya

    OpenAIRE

    Dellicour, Stephanie; Desai, Meghna; Mason, Linda; Odidi, Beatrice; Aol, George; Phillips-Howard, Penelope A.; Laserson, Kayla F.; ter Kuile, Feiko O.

    2013-01-01

    Background\\ud Understanding the socio-cultural context and perceptions of adverse pregnancy outcomes is important for informing the best approaches for public health programs. This article describes the perceptions, beliefs and health-seeking behaviours of women from rural western Kenya regarding congenital anomalies and miscarriages.\\ud \\ud Methods\\ud Ten focus group discussions (FGDs) were undertaken in a rural district in western Kenya in September 2010. The FGDs included separate groups c...

  6. Oral Cavity Squamous Cell Carcinoma - Characteristics and Survival in Aboriginal and Non-Aboriginal Western Australians

    OpenAIRE

    Frydrych, A.M; Slack-Smith, L.M; Parsons, R; Threlfall, T

    2014-01-01

    Background: Squamous cell carcinoma (SCC) is the most common type of malignancy affecting the oral cavity. While exposures to main risk factors for oral SCC such as smoking and alcohol use are higher amongst the Aboriginal people, little is known about oral cancer in this population. This study aimed to describe characteristics and survival of oral SCC in Aboriginal and non-Aboriginal Western Australians. Methods: All primary oral SCC cases reported to the Western Australian Cancer Registry (...

  7. Achieving Balance Through the Art of Eating: Demystifying Eastern Nutrition and Blending it with Western Nutrition

    OpenAIRE

    Shannon Wongvibulsin; Suzie Seoyang Lee; Ka-Kit Hui, MD, FACP

    2012-01-01

    Eastern and Western approaches to nutrition are unique and possess both strengths and weaknesses. Blending the best of both techniques will allow for the development of an integrative nutrition system that is more effective than either tradition on its own. The Western view to nutrition is already adopting certain attributes of the Eastern medicine philosophy as exemplified by the progression towards individualized nutrition through methods such as nutrigenetics. Nevertheless, many difference...

  8. Obesity and nutrition behaviours in Western and Palestinian outpatients with severe mental illness

    Directory of Open Access Journals (Sweden)

    Aljeesh Yousef

    2011-10-01

    Full Text Available Abstract Background While people with severe mental illness have been found to be more overweight and obese in Western nations, it is unknown to what extent this occurs in Middle Eastern nations and which eating behaviours contribute to obesity in Middle Eastern nations. Method A total of 665 responses were obtained from patients with serious mental illness attending out-patient clinics in Western developed countries (Germany, UK and Australia; n = 518 and Palestine (n = 147. Patients were evaluated by ICD-10 clinical diagnosis, anthropometric measurements and completed a self-report measure of frequencies of consuming different food items and reasons for eating. Nutritional habits were compared against a Western normative group. Results More participants from Palestine were overweight or obese (62% compared to Western countries (47%. In the Western sample, obese patients reported consuming more low-fat products (OR 2.54, 95% CI 1.02-6.33 but also greater eating due to negative emotions (OR 1.84, 95% CI 1.31-2.60 than patients with a healthy body-mass index. In contrast, obese patients from Palestine reported increased consumption of unhealthy snacks (OR 3.73 95% CI 1.16-12.00. Conclusion Patients with mental illness have poorer nutritional habits than the general population, particularly in Western nations. Separate interventions to improve nutritional habits and reduce obesity are warranted between Western nations and Palestine.

  9. New Method to Disaggregate and Analyze Single Isolated Helminthes Cells Using Flow Cytometry: Proof of Concept

    Directory of Open Access Journals (Sweden)

    Karen Nava-Castro

    2011-01-01

    Full Text Available In parasitology, particularly in helminthes studies, several methods have been used to look for the expression of specific molecules, such as RT-PCR, western blot, 2D-electrophoresis, and microscopy, among others. However, these methods require homogenization of the whole helminth parasite, preventing evaluation of individual cells or specific cell types in a given parasite tissue or organ. Also, the extremely high interaction between helminthes and host cells (particularly immune cells is an important point to be considered. It is really hard to obtain fresh parasites without host cell contamination. Then, it becomes crucial to determine that the analyzed proteins are exclusively from parasitic origin, and not a consequence of host cell contamination. Flow cytometry is a fluorescence-based technique used to evaluate the expression of extra-and intracellular proteins in different type cells, including protozoan parasites. It also allows the isolation and recovery of single-cell populations. Here, we describe a method to isolate and obtain purified helminthes cells.

  10. Pulsed Dilution Method for the Recovery of Aggregated Mouse TNF-α

    Directory of Open Access Journals (Sweden)

    Merat Mahmoodi

    2017-05-01

    Full Text Available Background: The expression of mouse tumor necrosis factor alpha (TNF-α in Escherichia coli is a favorable way to get high yield of protein; however, the formation of cytoplasmic inclusion bodies, which is the consequence of insoluble accumulated proteins, is a major obstacle in this system. To overcome this obstacle, we used a pulsed dilution method to convert the product to its native conformation. Methods: Reducing agent and guanidine hydrochloride were used to solubilize inclusion bodies formed after TNF-(α expression. Then, the refolding procedure was performed by pulsed dilution of the denatured protein into a refolding buffer. The properly-folded protein was purified by metal affinity chromatography. Results: SDS-PAGE showed a 19.9 kDa band related to the mature TNF-(α protein. The protein was recognized by anti-mouse TNF-(α on western blots. The final concentration of the purified recombinant TNF-(α was 62.5 μg/mL. Conclusions: Our study demonstrates the efficiency of this method to produce a high yield of folded mature TNF- (α.

  11. Long-term monitoring of western aspen--lessons learned.

    Science.gov (United States)

    Strand, E K; Bunting, S C; Starcevich, L A; Nahorniak, M T; Dicus, G; Garrett, L K

    2015-08-01

    Aspen woodland is an important ecosystem in the western United States. Aspen is currently declining in western mountains; stressors include conifer expansion due to fire suppression, drought, disease, heavy wildlife and livestock use, and human development. Forecasting of tree species distributions under future climate scenarios predicts severe losses of western aspen within the next 50 years. As a result, aspen has been selected as one of 14 vital signs for long-term monitoring by the National Park Service Upper Columbia Basin Network. This article describes the development of a monitoring protocol for aspen including inventory mapping, selection of sampling locations, statistical considerations, a method for accounting for spatial dependence, field sampling strategies, and data management. We emphasize the importance of collecting pilot data for use in statistical power analysis and semi-variogram analysis prior to protocol implementation. Given the spatial and temporal variability within aspen stem size classes, we recommend implementing permanent plots that are distributed spatially within and among stands. Because of our careful statistical design, we were able to detect change between sampling periods with desired confidence and power. Engaging a protocol development and implementation team with necessary and complementary knowledge and skills is critical for success. Besides the project leader, we engaged field sampling personnel, GIS specialists, statisticians, and a data management specialist. We underline the importance of frequent communication with park personnel and network coordinators.

  12. Explaining ecological clusters of maternal depression in South Western Sydney

    Science.gov (United States)

    2014-01-01

    Background The aim of the qualitative study reported here was to: 1) explain the observed clustering of postnatal depressive symptoms in South Western Sydney; and 2) identify group-level mechanisms that would add to our understanding of the social determinants of maternal depression. Methods Critical realism provided the methodological underpinning for the study. The setting was four local government areas in South Western Sydney, Australia. Child and Family practitioners and mothers in naturally occurring mothers groups were interviewed. Results Using an open coding approach to maximise emergence of patterns and relationships we have identified seven theoretical concepts that might explain the observed spatial clustering of maternal depression. The theoretical concepts identified were: Community-level social networks; Social Capital and Social Cohesion; "Depressed community"; Access to services at the group level; Ethnic segregation and diversity; Supportive social policy; and Big business. Conclusions We postulate that these regional structural, economic, social and cultural mechanisms partially explain the pattern of maternal depression observed in families and communities within South Western Sydney. We further observe that powerful global economic and political forces are having an impact on the local situation. The challenge for policy and practice is to support mothers and their families within this adverse regional and global-economic context. PMID:24460690

  13. Social policies, separation, and second birth spacing in Western Europe

    Directory of Open Access Journals (Sweden)

    Michaela Kreyenfeld

    2017-10-01

    Full Text Available Objective: This paper studies postseparation fertility behavior. The aim is to investigate whether, and if so how, separation affects second birth spacing in Western European countries. Methods: This analysis makes use of rich survey data from Belgium, France, Germany, Italy, Spain, and the United Kingdom, as well as from Finnish register data. We thus cover the behavior of a large proportion of the population of Western Europe. We also use descriptive measures, such as Kaplan‒Meier survival functions and cumulative incidence curves. In the multivariate analysis, we employ event history modeling to show how education relates to postseparation fertility behavior. Results: There are large differences in postseparation fertility behavior across European countries. For Spain and Italy, we find that only a negligibly small proportion of the population have a second child after separating from the other parent of the firstborn child. The countries with the highest proportion of second children with a new partner are the United Kingdom, Germany, and Finland. In all countries, separation after first birth leads to a sharp increase in the birth interval between first and second births. Contribution: Our study is a contribution to the demographic literature that aims at understanding birth spacing patterns in Western Europe. Furthermore, we draw attention to the role of postseparation policies in explaining country differences in fertility behavior in contemporary societies.

  14. Erosion by rain in the western Congo

    International Nuclear Information System (INIS)

    Ploey, J. de

    1967-01-01

    Vast expanses of the western part of central and southern Africa are covered with uniform, sandy formations of the Kalahari type. The topography of these areas and their present morphological characteristics are mainly the result of erosion by rain. Information on the hydrology of the surface waters in these areas is fairly limited and is insufficient to permit any conclusions regarding the way in which erosion by rain takes place. To obtain a better understanding of these phenomena, the author devised a series of experiments based on the use of 46 Sc-labelled radioactive sand. These experiments began at the beginning of the 1964/65 rainy season and are continuing. The experimental plot corresponds to convex and rectilinear portions of a hillside with a slope varying between 0 and 12 degrees. The vegetation consists of grassy savannah of substeppe appearance and secondary forests. Series of labelled samples were placed successively on the surface of the experimental plot and the erosive effect of rain was determined by measuring the residual concentrations after rainfall. Some samples were placed below a shield so as to eliminate the effects of splash and reveal the part played by runoff. Radiographic films were used to study the dispersion of labelled particles in the surrounding area. This radiographic method made it possible to determine the scale of erosion by splash for different rainfall conditions. The erosion diagrams obtained from these experiments show the correlations that exist between the intensity and duration of the rainfall and the erosion of the soil. Examination of the erosion diagrams and the shielded samples and analysis of the radiographs showed that erosion by rain on Kalahari ground covered with substeppe savannah is caused mainly by splash erosion and by dispersed, intermittent runoff. Sheet wash plays no part if the slope is less than 12 degrees. (author) [fr

  15. Corporatism In Western Europe: Current State And Prospects For Evolution

    Directory of Open Access Journals (Sweden)

    Р. Ja. Feldman

    2018-01-01

    Full Text Available The article conducts a political analysis of the Western European institutions of corporatism. The main task of the author is the study of the policy of harmonizing the interests of labor and capital (trade unions and employers’ associations, which is implemented in countries such as the UK, Germany, France, Italy, Norway, Sweden, Denmark etc. Dynamics of political processes unfolding in the space of Western Europe, suggests that the mechanisms of articulation and political representation of social and labour interests have significantly transformed over the past 30 years. The use of institutional and systemic approaches along with the empirical methods, leads to the conclusion that the most developed European countries are moving from the classical model of corporatism to a more pluralistic forms of interaction between the state, labour and capital. Social partnership as an instrument of collective bargaining between employees and employers is displaced from the political sphere to the sectoral and organizational levels. The typical institutions of democratic corporatism (tripartite commissions, socio-economic councils, etc., who played a crucial role in rebuilding post-war Europe, become rudimentary organs of the national political systems. In addition, there is a tendency to weaken the political influence of trade unions, who successfully struggled for the satisfaction of collective demands of workers in the beginning of XX century. Large multinational companies prefer to influence the political decision-making centers autonomously, ignoring the associative membership in the guild organizations. As a consequence, corporatist bargaining is being replaced by direct and indirect lobbying, Government Relations and election fundraising. When accounting for identified trends, the author presents a hypothesis that the evolution of corporatism in Western Europe will lead to its gradual degeneration. Taking into account the identified trends, the

  16. Developing Biomass Equations for Western Hemlock and Red Alder Trees in Western Oregon Forests

    Science.gov (United States)

    Krishna Poudel; Hailemariam Temesgen

    2016-01-01

    Biomass estimates are required for reporting carbon, assessing feedstock availability, and assessing forest fire threat. We developed diameter- and height-based biomass equations for Western hemlock (Tsuga heterophylla (Raf.) Sarg.) and red alder (Alnus rubra Bong.) trees in Western Oregon. A system of component biomass...

  17. Phenotypic and genetic characterization of western prairie clover collections from the western USA

    Science.gov (United States)

    Kishor Bhattarai; B. Shaun Bushman; Douglas A. Johnson; John G. Carman

    2010-01-01

    Few North American legumes are available for rangeland revegetation in the semiarid western United States. Western prairie clover (Dalea ornata [Douglas ex Hook.] Eaton & J. Wright) is a perennial legume with desirable forage characteristics and is distributed in the northern Great Basin, Snake River Basin, and southern Columbia Plateau. Understanding the...

  18. Communication Patterns in Adult-Infant Interactions in Western and Non-Western Cultures.

    Science.gov (United States)

    Keller, Heidi; And Others

    1988-01-01

    Analyzes the early communication structure in adult-child interactions with two- to six-month old babies in Western (West Germany, Greece) and non-Western (Yanomami, Trobriand) societies. Discusses universal international verbal and non-verbal structures reflecting intuitive parenting programs. (FMW)

  19. Achieving Balance Through the Art of Eating: Demystifying Eastern Nutrition and Blending it with Western Nutrition

    Science.gov (United States)

    Wongvibulsin, Shannon; Lee, Suzie Seoyang; Hui, Ka-Kit

    2012-01-01

    Eastern and Western approaches to nutrition are unique and possess both strengths and weaknesses. Blending the best of both techniques will allow for the development of an integrative nutrition system that is more effective than either tradition on its own. The Western view to nutrition is already adopting certain attributes of the Eastern medicine philosophy as exemplified by the progression towards individualized nutrition through methods such as nutrigenetics. Nevertheless, many differences still remain between Eastern and Western nutritional concepts. Becoming fluent in both Western and Eastern methodologies can ensure the extraction of the best from both techniques for the development of a comprehensive, systematic, and holistic nutritional approach to achieve optimal health. PMID:24716109

  20. Achieving Balance Through the Art of Eating: Demystifying Eastern Nutrition and Blending it with Western Nutrition

    Directory of Open Access Journals (Sweden)

    Shannon Wongvibulsin

    2012-01-01

    Full Text Available Eastern and Western approaches to nutrition are unique and possess both strengths and weaknesses. Blending the best of both techniques will allow for the development of an integrative nutrition system that is more effective than either tradition on its own. The Western view to nutrition is already adopting certain attributes of the Eastern medicine philosophy as exemplified by the progression towards individualized nutrition through methods such as nutrigenetics. Nevertheless, many differences still remain between Eastern and Western nutritional concepts. Becoming fluent in both Western and Eastern methodologies can ensure the extraction of the best from both techniques for the development of a comprehensive, systematic, and holistic nutritional approach to achieve optimal health.

  1. 2-tiered antibody testing for early and late Lyme disease using only an immunoglobulin G blot with the addition of a VlsE band as the second-tier test.

    Science.gov (United States)

    Branda, John A; Aguero-Rosenfeld, Maria E; Ferraro, Mary Jane; Johnson, Barbara J B; Wormser, Gary P; Steere, Allen C

    2010-01-01

    Standard 2-tiered immunoglobulin G (IgG) testing has performed well in late Lyme disease (LD), but IgM testing early in the illness has been problematic. IgG VlsE antibody testing, by itself, improves early sensitivity, but may lower specificity. We studied whether elements of the 2 approaches could be combined to produce a second-tier IgG blot that performs well throughout the infection. Separate serum sets from LD patients and control subjects were tested independently at 2 medical centers using whole-cell enzyme immunoassays and IgM and IgG immunoblots, with recombinant VlsE added to the IgG blots. The results from both centers were combined, and a new second-tier IgG algorithm was developed. With standard 2-tiered IgM and IgG testing, 31% of patients with active erythema migrans (stage 1), 63% of those with acute neuroborreliosis or carditis (stage 2), and 100% of those with arthritis or late neurologic involvement (stage 3) had positive results. Using new IgG criteria, in which only the VlsE band was scored as a second-tier test among patients with early LD (stage 1 or 2) and 5 of 11 IgG bands were required in those with stage 3 LD, 34% of patients with stage 1, 96% of those with stage 2, and 100% of those with stage 3 infection had positive responses. Both new and standard testing achieved 100% specificity. Compared with standard IgM and IgG testing, the new IgG algorithm (with VlsE band) eliminates the need for IgM testing; it provides comparable or better sensitivity, and it maintains high specificity.

  2. Regional competitiveness: The case of Western China

    Directory of Open Access Journals (Sweden)

    Vuković Darko

    2010-01-01

    Full Text Available This paper explains the concept of regional competitiveness and the factors that influence on it. A large number of various authors explain this concept, based on its different aspects, including: productivity, mikroaspekts (firm, quality of human capital, innovation, technology, infrastructure, social capital, etc.. Taking into account complex nature of regional competitiveness, it is difficult to determine a standard definition of this term. The second part of this paper refers to the case of western China. Substantial disparity in regional development is a reality in every geographically large country, and the causes of the disparity are numerous and complex. Regional inequality has been an important issue in China. This paper generally summarized China's Western regions geography, government policies and development situation. The authors put forward some practical strategies on how to help the western regions create a favorable environment to attract national and international investment.

  3. 2012 Annual Report [WESTERN AREA POWER ADMINISTRATION

    Energy Technology Data Exchange (ETDEWEB)

    None

    2012-01-01

    Fiscal Year 2012 brought some tumultuous and uncertain times to Western. The utility industry and technology continued to evolve, and the demand for constant flow of power and transmission system reliability continued to increase. Western kept pace by continuing to deliver reliable, cost-based hydropower while reviewing and updating business practices that took into account how the energy industry is evolving. During this time of exponential change, Western tackled many challenges, including: Reviewing the Transmission Infrastructure Program processes and procedures; Responding to Secretary of Energy Steven Chu’s memorandum to create a modern, efficient and reliable transmission grid; Weathering record-breaking natural disasters in our service territory; Completing our role in TIP’s flagship project—the Montana Alberta Tie Ltd. transmission line; Incorporating new, far-reaching regulations and industry trends.

  4. Upper mantle flow in the western Mediterranean

    Energy Technology Data Exchange (ETDEWEB)

    Panza, G F [Dipartimento di Scienze della Terra, Universita degli Studi di Trieste, Trieste (Italy) and Abdus Salam International Centre for Theoretical Physics, Trieste (Italy); Raykova, R [Geophysical Institute of BAS, Sofia (Bulgaria) and Istituto Nazionale di Geofisica e Vulcanologia, Sezione Bologna, Bologna (Italy); Carminati, E; Doglioni, C [Dipartimento di Scienze della Terra, Universita degli Studi di Trieste, Trieste (Italy)

    2006-07-15

    Two cross-sections of the western Mediterranean Neogene-to-present backarc basin are presented, in which geological and geophysical data of the Transmed project are tied to a new shear-wave tomography. Major results are i) the presence of a well stratified upper mantle beneath the older African continent, with a marked low-velocity layer between 130-200 km of depth; ii) the dilution of this layer within the younger western Mediterranean backarc basin to the north, and iii) the easterly raising of a shallower low-velocity layer from about 140 km to about 30 km in the Tyrrhenian active part of the backarc basin. These findings suggest upper mantle circulation in the western Mediterranean backarc basin, mostly easterly-directed and affecting the boundary between upper asthenosphere (LVZ) and lower asthenosphere, which undulates between about 180 km and 280 km. (author)

  5. Upper mantle flow in the western Mediterranean

    International Nuclear Information System (INIS)

    Panza, G.F.; Raykova, R.; Carminati, E.; Doglioni, C.

    2006-07-01

    Two cross-sections of the western Mediterranean Neogene-to-present backarc basin are presented, in which geological and geophysical data of the Transmed project are tied to a new shear-wave tomography. Major results are i) the presence of a well stratified upper mantle beneath the older African continent, with a marked low-velocity layer between 130-200 km of depth; ii) the dilution of this layer within the younger western Mediterranean backarc basin to the north, and iii) the easterly raising of a shallower low-velocity layer from about 140 km to about 30 km in the Tyrrhenian active part of the backarc basin. These findings suggest upper mantle circulation in the western Mediterranean backarc basin, mostly easterly-directed and affecting the boundary between upper asthenosphere (LVZ) and lower asthenosphere, which undulates between about 180 km and 280 km. (author)

  6. Immunogenicity of Bothrops atrox (Ophidia: Viperidae venom and its evaluation by immunoenzymatic methods Inmunogenicidad del veneno de Bothrops atrox (Ophidia: Viperidae y su evaluación por métodos inmunoenzimáticos

    Directory of Open Access Journals (Sweden)

    Gustavo A. Sandoval

    2012-02-01

    Full Text Available The immunogenicity of Bothrops atrox, “jergón”, venom was studied using ELISA and Western Blot methods, as well as cross-reactivity patterns against venoms of Bothrops brazili, Lachesis muta and Crotalus durissus. For this purpose, New Zealand white rabbits (2 kg aprox were immunized with four 500 μg doses of B. atrox venom in a period of 90 days. Antibody production was followed using ELISA technique, and title of hiper-immune serum was determined at the end of immunization protocol. Additionally, electrophoretic patterns of venoms were analyzed by SDS-PAGE and venom reactivity against obtained serum by ELISA and Western Blot. Immunization schedule allowed a pronounced antibody production since day 20 of protocol. At the end of process, serum title was 256000, which demonstrated both efficacy and usefulness of the developed procedure. On the other hand, studied venoms showed a heterogenic protein composition according to their electrophoretic patterns, whereas cross-reactivity values of 23,7%, 4,0% and 1,8% were obtained between B. atrox venom and B. brazili, L. muta and C. durissus venoms, respectively, using immunoenzymatic methods. According to our results, this procedure constitutes an initial step for further assays directed to optimization in immunoserum production for envenoming treatment and development of kits for diagnosis and species identification of snakes.Se estudió la inmunogenicidad del veneno de la serpiente Bothrops atrox, “jergón”, utilizando los métodos inmunoenzimáticos de ELISA y Western Blot, así como los patrones de reactividad cruzada empleando los venenos de las serpientes Bothrops brazili, Lachesis muta y Crotalus durissus. Para este fin se inmunizaron conejos albinos Nueva Zelanda (2 kg aprox con cuatro dosis de 500 μg del veneno de B. atrox en un periodo de 90 días. La producción de anticuerpos fue monitoreada mediante la técnica de ELISA, determinándose el título del suero hiperinmune obtenido

  7. Bioethical concerns are global, bioethics is Western

    Science.gov (United States)

    Chattopadhyay, Subrata; De Vries, Raymond

    2009-01-01

    Modern bioethics was born in the West and thus reflects, not surprisingly, the traditions of Western moral philosophy and political and social theory. When the work of bioethics was confined to the West, this background of socio-political theory and moral tradition posed few problems, but as bioethics has moved into other cultures - inside and outside of the Western world - it has become an agent of moral imperialism. We describe the moral imperialism of bioethics, discuss its dangers, and suggest that global bioethics will succeed only to the extent that it is local. PMID:19593391

  8. Oil and gas competition in Western Europe

    International Nuclear Information System (INIS)

    Carrie, J.L.

    1991-01-01

    This paper reports that in Western Europe, the competition between oil and gas began on a large scale during the 1960s. Indeed, natural gas accounted for only 2 percent of Western Europe's energy consumption in 1960 and for already 7 percent in 1970. It now accounts for about 17 percent. Almost all of this increase took place at the detriment of oil products. The competition between those two energy sources has resulted in the development of natural gas supply on one hand, and in several political and economic factors on the other

  9. AN INNER STRUGGLE: UNDER WESTERN EYES

    Directory of Open Access Journals (Sweden)

    Hale Yağlıdere

    2013-06-01

    Full Text Available This study aims to shed light on the most significant aspect of the modern man’s inner struggle in Joseph Conrad’s Under Western Eyes. Joseph Conrad is one of the authors of the twentieth century who was influenced by the psychological philosophy and he makes use of this influence in his works deeply. He who synthesizes the inner nature of the modern man, having conflicts himself with the deeper analysis field of the psychology aims to point out the incomprehensible and unsettled inner nature of the modern man in Under Western Eyes.

  10. Middle Class Fortunes in Western Europe

    OpenAIRE

    Kochhar, Rakesh

    2017-01-01

    This paper examines the state of the middle classes in the U.S. and 11 countries in Western Europe and how it has changed since 1991. Among Western Europe's six largest economies, the shares of adults living in middle-income households increased in France, the Netherlands and the United Kingdom from 1991 to 2010, but shrank in Germany, Italy and Spain. France, the Netherlands and the UK also experienced notable growth in disposable household income, but incomes were either stagnant or falling...

  11. Bioethical concerns are global, bioethics is Western.

    Science.gov (United States)

    Chattopadhyay, Subrata; De Vries, Raymond

    2008-07-01

    Modern bioethics was born in the West and thus reflects, not surprisingly, the traditions of Western moral philosophy and political and social theory. When the work of bioethics was confined to the West, this background of socio-political theory and moral tradition posed few problems, but as bioethics has moved into other cultures - inside and outside of the Western world - it has become an agent of moral imperialism. We describe the moral imperialism of bioethics, discuss its dangers, and suggest that global bioethics will succeed only to the extent that it is local.

  12. 75 FR 5760 - Proposed Information Collection; Comment Request; Western Alaska Community Development Quota Program

    Science.gov (United States)

    2010-02-04

    ... before April 5, 2010. ADDRESSES: Direct all written comments to Diana Hynek, Departmental Paperwork... and diversified local economies in western Alaska. CDQ and prohibited species quota (PSQ) allocations... collection. II. Method of Collection Respondents have a choice of either electronic or paper forms. Methods...

  13. Microbial dolomite crusts from the carbonate platform off western India

    Digital Repository Service at National Institute of Oceanography (India)

    Rao, V.P.; Kessarkar, P.M.; Krumbein, W.E.; Krajewski, K.P.; Schneider, R.J.

    Association of Sedimentologists, Sedimentology, 50, 819-830 Microbial dolomite crusts off western India 821 dolomite crusts [2 x 2·5 em to 3 x 0'5 em) and pebbles occur within these sediments. METHODS Thin sections of the crusts were studied petro graphically....E. (19S7) Holocene dolomitization Df supr

  14. Peabody Western Coal cuts costs with bottom-dump haulers

    Energy Technology Data Exchange (ETDEWEB)

    Perla, S.; Baecker, G.; Morgan, W. [Empire Machinery, Mesa, AZ (United States)

    1995-04-01

    A new hauling concept has been introduced at the Black Mesa and Kayenta coal mines of the Peabody Western Coal Co. in northern Arizona, USA. The article describes the switch from Caterpillar 992 wheel loaders with 136 t bottom-dump trucks to 272 t bottom-dump trucks. Cat 789 off-highway trucks were modified to pull bottom-dump trucks. Haulage costs per ton of coal and cost per ton-mile have fallen significantly since the introduction of the new large hauling method. 7 figs., 2 photos.

  15. Chemical Composition and In Vitro Cytotoxicity of Essential Oils from Leaves and Flowers of Callistemon citrinus from Western Himalayas

    Science.gov (United States)

    Kumar, Dharmesh; Sukapaka, Mahesh; Babu, G. D. Kiran; Padwad, Yogendra

    2015-01-01

    Background Plant-based traditional system of medicine continues to play an important role in healthcare. In order to find new potent source of bioactive molecules, we studied the cytotoxic activity of the essential oils from the flowers and leaves of Callistemon citrinus. This is the first report on anticancer potential of essential oils of C. citrinus. Methods Cytotoxicity of essential oil was evaluated using sulfo-rhodamine B (SRB) assay against human lung carcinoma (A549), rat glioma (C-6), human colon cancer (Colo-205) and human cervical cancer (SiHa) cells. Apoptosis induction was evaluated by caspase-3/7 activity which was further confirmed by western blotting. Percentage cell apoptosis was determined by Annexin V based dead cell assay followed by DNA content as cell cycle analysis against A549 and C-6 cells. While 3-(4,5-dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay was used to check the toxicity against normal human peripheral blood mononuclear cells (PBMCs), the immunomodulatory activity on mouse splenocytes was evaluated using SRB assay. Results The GC and GC-MS analysis of these essential oils revealed high content of α-pinene (32.3%), limonene (13.1%) and α-terpineol (14.6%) in leaf sample, whereas the flower oil was dominated by 1,8-cineole (36.6%) followed by α-pinene (29.7%). The leaf oil contained higher amount of monoterpene hydrocarbons (52.1%) and sesquiterpenoids (14%) as compared to flower oil (44.6% and 1.2%, respectively). However, the flower oil was predominant in oxygenated monoterpenes (43.5%). Although both leaf and flower oils showed highest cytotoxicity on A549 cells (61.4%±5.0 and 66.7%±2.2, respectively), only 100 μg/mL flower oil was significantly active against C-6 cells (69.1%±3.1). Interestingly, no toxicity was recorded on normal cells. Conclusion Higher concentration of 1,8-cineole and/or synergistic effect of the overall composition were probably responsible for the efficacy of flower and leaf oils

  16. A Novel Method of Imaging Lysosomes in Living Human Mammary Epithelial Cells

    Directory of Open Access Journals (Sweden)

    Kristine Glunde

    2003-01-01

    Full Text Available Cancer cells invade by secreting degradative enzymes which, under normal conditions, are sequestered in lysosomal vesicles. The ability to noninvasively label lysosomes and track lysosomal trafficking would be extremely useful to understand the mechanisms by which degradative enzymes are secreted in the presence of pathophysiological environments, such as hypoxia and acidic extracellular pH, which are frequently encountered in solid tumors. In this study, a novel method of introducing a fluorescent label into lysosomes of human mammary epithelial cells (HMECs was evaluated. Highly glycosylated lysosomal membrane proteins were labeled with a newly synthesized compound, 5-dimethylamino-naphthalene-1-sulfonic acid 5-amino-3,4,6-trihydroxy-tetrahydro-pyran-2-ylmethyl ester (6-O-dansyl-GlcNH2. The ability to optically image lysosomes using this new probe was validated by determining the colocalization of the fluorescence from the dansyl group with immunofluorescent staining of two well-established lysosomal marker proteins, LAMP-1 and LAMP-2. The location of the dansyl group in lysosomes was also verified by using an anti-dansyl antibody in Western blots of lysosomes isolated using isopycnic density gradient centrifugation. This novel method of labeling lysosomes biosynthetically was used to image lysosomes in living HMECs perfused in a microscopy-compatible cell perfusion system.

  17. A Time- and Cost-Saving Method of Producing Rat Polyclonal Antibodies

    International Nuclear Information System (INIS)

    Wakayama, Tomohiko; Kato, Yukio; Utsumi, Rie; Tsuji, Akira; Iseki, Shoichi

    2006-01-01

    Producing antibodies usually takes more than three months. In the present study, we introduce a faster way of producing polyclonal antibodies based on preparation of the recombinant oligopeptide as antigen followed by immunization of rats. Using this method, we produced antisera against two mouse proteins: ERGIC-53 and c-Kit. An expression vector ligated with a pair of complementary synthetic oligodeoxyribonucleotides encoding the protein was introduced into bacteria, and the recombinant oligopeptide fused with the carrier protein glutathione-S-transferase was purified. Wistar rats were immunized by injecting the emulsified antigen subcutaneously into the hind footpads, followed by a booster injection after 2 weeks. One week after the booster, the sera were collected and examined for the antibody titer by immunohistochemistry. Antisera with 1600-fold titer at the maximum were obtained for both antigens and confirmed for their specificity by Western blotting. Anti-ERGIC-53 antisera recognized acinar cells in the sublingual gland, and anti-c-Kit antisera recognized spermatogenic and Leydig cells in the testis. These antisera were applicable to fluorescent double immunostaining with mouse monoclonal or rabbit polyclonal antibodies. Consequently, this method enabled us to produce specific rat polyclonal antisera available for immunohistochemistry in less than one month at a relatively low cost

  18. Western Option - Disarmament of Russian Weapon Plutonium

    International Nuclear Information System (INIS)

    Tveiten, B.; Petroll, M.R.

    2002-01-01

    The Western Option concept describes an approach to the conversion of weapon-grade plutonium from Russian nuclear warheads under the special aspects of meeting the criteria of irreversible utilization. Putting this concept of plutonium conversion into non-weapon-grade material into effect would make a major contribution to improving security worldwide. This study is based on an agreement between the Russian Federation and the United States of America concluded in September 2000. It provides for the conversion of 34 t of weapon-grade plutonium in each of the two states. This goal is also supported by other G8 countries. While the United States performs its part of the agreement under its sole national responsibility, the Russian program needs financial support by Western states. Expert groups have pointed out several options as a so-called basic scenario. The funds of approx. US Dollar 2 billion required to put them into effect have not so far been raised. The Western Option approach described in this contribution combines results of the basic scenario with other existing experience and with technical solutions available for plutonium conversion. One of the attractions of the Western Option lies in its financial advantages, which are estimated to amount to approx. US Dollar 1 billion. (orig.) [de

  19. Geothermal Energy Potential in Western United States

    Science.gov (United States)

    Pryde, Philip R.

    1977-01-01

    Reviews types of geothermal energy sources in the western states, including hot brine systems and dry steam systems. Conversion to electrical energy is a major potential use of geothermal energy, although it creates environmental disruptions such as noise, corrosion, and scaling of equipment. (AV)

  20. Identity and Islamic Radicalization in Western Europe

    NARCIS (Netherlands)

    S.M. Murshed (Syed); S. Pavan (Sara)

    2009-01-01

    textabstractThis paper argues that both socio-economic disadvantage and political factors, such as the West’s foreign policy with regard to the Muslim world, along with historical grievances, play a part in the development of Islamic radicalized collective action in Western Europe. We emphasise the

  1. Western hemlock as a veneer resource.

    Science.gov (United States)

    Thomas D. Fahey; Jr. Woodfin

    1982-01-01

    Presents recovery of veneer grade and volume from western hemlock from Oregon and Washington. Veneer grade recovery varied by grade and size of logs. Veneer volume recovered was about 45 percent of the cubic volume of the log and varied somewhat with log diameter.

  2. Stumpage market integration in western national forests

    Science.gov (United States)

    Jean M. Daniels

    2011-01-01

    This study presents results of statistical tests for stumpage market integration on 62 national forests in the Western United States. Quarterly stumpage prices from 1984 to 2007 obtained from cut and sold reports for USDA Forest Service Regions 1, 4, 5, and 6 (Northern, Intermountain, Pacific Southwest, and Pacific Northwest, respectively) were analyzed to establish...

  3. When science became Western: historiographical reflections.

    Science.gov (United States)

    Elshakry, Marwa

    2010-03-01

    While thinking about the notion of the "global" in the history of the history of science, this essay examines a related but equally basic concept: the idea of "Western science." Tracing its rise in the nineteenth century, it shows how it developed as much outside the Western world as within it. Ironically, while the idea itself was crucial for the disciplinary formation of the history of science, the global history behind this story has not been much attended to. Drawing on examples from nineteenth-century Egypt and China, the essay begins by looking at how international vectors of knowledge production (viz., missionaries and technocrats) created new global histories of science through the construction of novel genealogies and through a process of conceptual syncretism. Turning next to the work of early professional historians of science, it shows how Arabic and Chinese knowledge traditions were similarly reinterpreted in light of the modern sciences, now viewed as part of a diachronic and universalist teleology ending in "Western science." It concludes by arguing that examining the global emergence of the idea of Western science in this way highlights key questions pertaining to the relation of the history of science to knowledge traditions across the world and the continuing search for global histories of science.

  4. Potato pathogens in Northern and Western Europe

    NARCIS (Netherlands)

    Schepers, H.T.A.M.; Cooke, L.R.

    2015-01-01

    The climatic conditions in Northern and Western (NW) Europe, which are very suitable for the cultivation of potatoes, are also very favourable for a range of diseases of which late blight, caused by Phytophthora infestans, is by far the most important with the potential to cause total crop loss. It

  5. Indicators of cull in western Oregon conifers.

    Science.gov (United States)

    Paul E. Aho

    1982-01-01

    Descriptions and color photographs of important fungal sporophores (conks), other indicators of cull (wounds), and associated decays in western Oregon conifers are provided to aid timber markers, cruisers, and scalers in identifying them. Cull factors are given for the indicators by tree species.

  6. Capitalism in Six Westerns by John Ford

    Science.gov (United States)

    Braun, Carlos Rodriguez

    2011-01-01

    The economic and institutional analysis of capitalism can be illustrated through John Ford's Westerns. This article focuses on six classics by Ford that show the move toward modern order, the creation of a new society, and the rule of law. Economic features are pervading, from property rights and contracts to markets, money, and trade. Ford has…

  7. Medical humanities as expressive of Western culture.

    Science.gov (United States)

    Hooker, Claire; Noonan, Estelle

    2011-12-01

    In this paper we articulate a growing awareness within the field of the ways in which medical humanities could be deemed expressive of Western cultural values. The authors suggest that medical humanities is culturally limited by a pedagogical and scholarly emphasis on Western cultural artefacts, as well as a tendency to enact an uncritical reliance upon foundational concepts (such as 'patient' and 'experience') within Western medicine. Both these tendencies within the field, we suggest, are underpinned by a humanistic emphasis on appreciative or receptive encounters with 'difference' among patients that may unwittingly contribute to the marginalisation of some patients and healthcare workers. While cultural difference should be acknowledged as a central preoccupation of medical humanities, we argue that the discipline must continue to expand its scholarly and critical engagements with processes of Othering in biomedicine. We suggest that such improvements are necessary in order to reflect the cultural diversification of medical humanities students, and the geographical expansion of the discipline within non-Western and/or non-Anglophone locations.

  8. Asian and western Intellectual Capital in encounter

    NARCIS (Netherlands)

    Marien van den Boom; Dr. Daan Andriessen

    2007-01-01

    The purpose of the paper is to start a dialogue about differences between Western and Eastern cultures in the way they conceptualize knowledge and discuss the implications of these differences for a global intellectual capital (IC) theory and practice. A systematic metaphor analysis of the concept

  9. Estimating Western U.S. Reservoir Sedimentation

    Science.gov (United States)

    Bensching, L.; Livneh, B.; Greimann, B. P.

    2017-12-01

    Reservoir sedimentation is a long-term problem for water management across the Western U.S. Observations of sedimentation are limited to reservoir surveys that are costly and infrequent, with many reservoirs having only two or fewer surveys. This work aims to apply a recently developed ensemble of sediment algorithms to estimate reservoir sedimentation over several western U.S. reservoirs. The sediment algorithms include empirical, conceptual, stochastic, and processes based approaches and are coupled with a hydrologic modeling framework. Preliminary results showed that the more complex and processed based algorithms performed better in predicting high sediment flux values and in a basin transferability experiment. However, more testing and validation is required to confirm sediment model skill. This work is carried out in partnership with the Bureau of Reclamation with the goal of evaluating the viability of reservoir sediment yield prediction across the western U.S. using a multi-algorithm approach. Simulations of streamflow and sediment fluxes are validated against observed discharges, as well as a Reservoir Sedimentation Information database that is being developed by the US Army Corps of Engineers. Specific goals of this research include (i) quantifying whether inter-algorithm differences consistently capture observational variability; (ii) identifying whether certain categories of models consistently produce the best results, (iii) assessing the expected sedimentation life-span of several western U.S. reservoirs through long-term simulations.

  10. Amphibian distribution patterns in western Europe

    NARCIS (Netherlands)

    Zuiderwijk, Annie

    1980-01-01

    Mechanisms controlling the distribution of amphibians in western Europe have been studied in France where related species, isolated from each other at least during the last glacial period, are now sympatric. Occurrences and biotope preferences of the various species were investigated in several

  11. Social anxiety in three Western societies.

    NARCIS (Netherlands)

    Dam-Baggen, R.M.J. van; Kraaimaat, F.W.; Elal, G.

    2003-01-01

    The present study investigates whether empirical data support the notion that people in Western societies do not differ with regard to social anxiety. Social anxiety in Dutch students (N = 425) was compared with that experienced by students in the United States (N = 440) and Turkey (N = 349). Social

  12. Miocene freshwater Mollusca from western Brazilian Amazonia

    NARCIS (Netherlands)

    Wesselingh, F.P.; Ranzi, A.; Räsänen, M.E.

    2006-01-01

    Thirteen species of fossil molluscs are reported from the Solimões Formation of western Brazilian Amazonia. Based on mammalian chronology of the Solimões Formation and radiometric ages reported from coeval deposits in adjacent Peru, the age of the fauna is established as Late Miocene. The fauna

  13. Rising synchrony controls western North American ecosystems

    Science.gov (United States)

    Bryan A. Black; Peter van der Sleen; Emanuele Di Lorenzo; Daniel Griffin; William J. Sydeman; Jason B. Dunham; Ryan R. Rykaczewski; Marisol García-Reyes; Mohammad Safeeq; Ivan Arismendi; Steven J. Bograd

    2018-01-01

    Along the western margin of North America, the winter expression of the North Pacific High (NPH) strongly influences interannual variability in coastal upwelling, storm track position, precipitation, and river discharge. Coherence among these factors induces covariance among physical and biological processes across adjacent marine and terrestrial ecosystems. Here, we...

  14. Eghindi among Sahrawi refugees of Western Sahara

    NARCIS (Netherlands)

    Volpato, G.

    2014-01-01

    Eghindi is an illness built around a set of pathological states experienced by Sahrawi in the desert environment of Western Sahara. Its core symptoms are caused by osmotic imbalances related to salt consumption. In 1975, many Sahrawi were exiled into refugee camps, and they have since experienced

  15. Parenting in non-Western migrant families

    NARCIS (Netherlands)

    Freek Bucx; Simone de Roos

    2015-01-01

    Original Title: Opvoeden in niet-westerse migrantengezinnen This report describes the parenting of young children in families of non-Western origin. The focus is mainly on parents and children of Turkish, Moroccan, Surinamese and Antillean origin. Based on earlier qualitative research and

  16. Women's and Feminist Activism in Western Europe

    NARCIS (Netherlands)

    Buikema, R.L.

    First- and second-wave Western European feminists struggled to realize full access to civil rights for women and the creation of a participatory democracy that ensured social solidarity. They consequently stressed the fact that in addition to the struggle for civil rights, women needed to contest

  17. Western cratonic domain in Uruguay: geochronology

    International Nuclear Information System (INIS)

    Preciozzi, F.; Peel, F.; Muzio, R.; Ledesma, J.; Guerequiz, R.

    2009-01-01

    In this article has been studied the Western cratonic in Uruguay are divided into three major units: Piedra Alta Terrane, Valentines Block and Pavas Terrane. Piedra Alta Terrane has of evidence of Neo proterozoic orogenesis . Sarandi del Yi -Arroyo Solis Grande shear zone separate, it from Valentine block . Valentine Block separate it from Pavas terrane by Cueva del Tigre shear zone

  18. Nickel-accumulating plant from Western Australia

    Energy Technology Data Exchange (ETDEWEB)

    Severne, B C; Brooks, R R

    1972-01-01

    A small shrub Hybanthus floribundus (Lindl.) F. Muell. Violaceae growing in Western Australia accumulates nickel and cobalt to a very high degree. Values of up to 23% nickel in leaf ash may represent the highest relative accumulation of a metal on record. The high accumulation of nickel poses interesting problems in plant physiology and plant biochemistry. 9 references, 2 figures, 1 table.

  19. Western European Art Foundations and Publishing

    Science.gov (United States)

    King, Lindsay M.; Clement, Russell T.

    2011-01-01

    Western European art foundations create invaluable opportunities for research and exhibition by artists, curators, and scholars. These activities are often documented and disseminated via high-quality publications. This article highlights an important but under-recognized collecting resource for academic and museum libraries by profiling several…

  20. Corporate actors in Western European television news

    NARCIS (Netherlands)

    Verhoeven, P.

    2009-01-01

    News about corporations can be understood as an interdependent relationship among the public relations function, organizational logic and the logic of the media. This research addresses the visibility and role of corporate actors in Western European public television news. A quantitative content