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Sample records for western blot study

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

    Science.gov (United States)

    Hall, Randy A

    2015-01-01

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

  2. The western blot

    Science.gov (United States)

    Western blotting is a technique that involves the separation of proteins by gel electrophoresis, their blotting or transfer to a membrane, and selective immunodetection of an immobilized antigen. This is an important and routine method for protein analysis that depends on the specificity of antibod...

  3. Western blot analysis.

    Science.gov (United States)

    Hirano, Seishiro

    2012-01-01

    Electrophoresis and the following western blot analysis are indispensable to investigate biochemical changes in cells and tissues exposed to nanoparticles or nanomaterials. Proteins should be extracted from the cells and tissues using a proper method, especially when phosphorylated proteins are to be detected. It is important to select a good blocking agent and an appropriate pair of primary and peroxidase-tagged secondary antibodies to obtain good results in western blot analysis. One thing that may be specific to nanomaterials, and that you should keep in mind, is that some proteins may be adsorbed on the surface of particulate nanomaterials. In this chapter the whole process of western blot analysis, from sample preparation to quantitative measurement of target proteins, is described.

  4. Western Blot Techniques.

    Science.gov (United States)

    Kim, Brianna

    2017-01-01

    The Western blot is an important laboratory technique that allows for specific identification and characterization of proteins. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)-separated proteins are electophoretically transferred to a polyvinylidene fluoride (PVDF) membrane which is then incubated with specific antibodies, then developed to show the protein of interest. Here, we describe the transfer and detection of Outer surface protein A (OspA), a protein only found on the surface of Borrelia burgdorferi, the bacteria responsible for Lyme disease.

  5. Western Blot Analysis of C. elegans Proteins.

    Science.gov (United States)

    Jeong, Dae-Eun; Lee, Yujin; Lee, Seung-Jae V

    2018-01-01

    C. elegans has been widely used as a model organism for basic biological research and is particularly amenable for molecular genetic studies using a broad repertoire of techniques. Biochemical approaches, including Western blot analysis, have emerged as a powerful tool in C. elegans biology for understanding molecular mechanisms that link genotypes to phenotypes. Here, we provide a protocol for Western blot analysis using protein extracts obtained from C. elegans samples.

  6. Autophagy and Proteases: Basic Study of the Autophagic Flux by Western Blot.

    Science.gov (United States)

    Fernández, Álvaro F

    2018-01-01

    Autophagy is a catabolic process triggered in the cell by a wide range of stress stimuli, both external (including nutrient deprivation) and internal (like the presence of protein aggregates or damaged organelles). First described in yeast, this pathway has recently gained major importance due to its role in several pathologies, from inflammatory processes to cancer or aging. However, its analysis can be easily misinterpreted if it is not done properly, leading to conflicting results. Here, the classical autophagy flux study by Western blot is described, as a first and basic analysis of the status of autophagy in a given system.

  7. A Study of Rubisco through Western Blotting and Tissue Printing Techniques

    Science.gov (United States)

    Ma, Zhong; Cooper, Cynthia; Kim, Hyun-Joo; Janick-Buckner, Diane

    2009-01-01

    We describe a laboratory exercise developed for a cell biology course for second-year undergraduate biology majors. It was designed to introduce undergraduates to the basic molecular biology techniques of Western blotting and immunodetection coupled with the technique of tissue printing in detecting the presence, relative abundance, and…

  8. Lectin-probed western blot analysis.

    Science.gov (United States)

    Sato, Takeshi

    2014-01-01

    Lectin-probed western blot analysis, the so-called lectin blot analysis, is a useful method to yield basic information on the glycan structures of glycoproteins, based on the carbohydrate-binding specificities of lectins. By lectin blot analysis, researchers can directly analyze the glycan structures without releasing the glycans from glycoproteins. Here, the author describes protocols for standard analysis, and applies analysis in combination with glycosidase digestion of blot.

  9. Western blot: technique, theory, and trouble shooting.

    Science.gov (United States)

    Mahmood, Tahrin; Yang, Ping-Chang

    2012-09-01

    Western blotting is an important technique used in cell and molecular biology. By using a western blot, researchers are able to identify specific proteins from a complex mixture of proteins extracted from cells. The technique uses three elements to accomplish this task: (1) separation by size, (2) transfer to a solid support, and (3) marking target protein using a proper primary and secondary antibody to visualize. This paper will attempt to explain the technique and theory behind western blot, and offer some ways to troubleshoot.

  10. Risk factors for repeatedly reactive HIV-1 EIA and indeterminate western blots. A population-based case-control study.

    Science.gov (United States)

    Celum, C L; Coombs, R W; Jones, M; Murphy, V; Fisher, L; Grant, C; Corey, L; Inui, T; Wener, M H; Holmes, K K

    1994-05-23

    Causes of indeterminate results of Western blot testing (IWB) for human immunodeficiency virus (HIV) type 1 include seroconversion, HIV-2 cross-reactivity, and autoimmune disease, but most IWB results remain unexplained. This case-control study assessed risk factors for IWB results, including early HIV infection, other retroviral infection, autoantibodies, and other medical conditions. Prospective study to determine HIV seroconversion rate, with a case-control design to assess other risk factors for IWB. Cases (persons with one or more repeatedly reactive HIV-1 enzyme immunoassay with IWB), their current sexual partners, and controls (persons with negative enzyme immunoassay and Western blot results) were recruited from blood banks, health department and prenatal clinics, and private providers in Washington and Oregon. Of 244 cases enrolled, 206 were followed up for 6 months or longer, and six (3.0%; 95% confidence interval [CI], 0.7% to 5.3%) with recent HIV risk behaviors seroconverted. The Western blot banding patterns differed among groups; cases usually had p17 or p24 bands, while controls and cases' sexual partners usually had polymerase bands. Conditional logistic regression indicated that independent risk factors for IWB among male cases and controls were a tetanus booster in the past 2 years (odds ratio, 3.2; 95% CI, 1.2 to 8.6) and sexual contact with a prostitute (odds ratio, 3.0; 95% CI, 1.0 to 9.5). Independent risk factors for women were parity (odds ratio, 1.2; 95% CI, 1.02 to 1.4) and autoantibodies, either rheumatoid factor or antinuclear antibodies (odds ratio, 2.3; 95% CI, 1.03 to 5.6). No cross-reactivity was detected with HIV-2, human T-lymphotrophic virus type 1, feline immunodeficiency or feline leukemia, or bovine immunodeficiency viruses. Evaluation of persons with reactive HIV-1 enzyme immunoassays and IWB should include an assessment of HIV risk and other possible risk factors, such as alloimmunization (ie, parity or recent immunization

  11. Interpretation criteria in Western blot diagnosis of Lyme borreliosis.

    Science.gov (United States)

    Mavin, S; McDonagh, S; Evans, R; Milner, R M; Chatterton, J M W; Ho-Yen, D O

    2011-01-01

    This study reviews the Lyme borreliosis Western blot interpretation process, including what bands are classed as specific, the number of bands needed for a positive result, the role of band intensity and the use of clinical information. In 2008, 3688 patients (4223 serum samples) were tested by enzyme immunoassay (EIA), with 832 patients tested by confirmatory in-house IgG Western blot: 272 patients were Western blot-positive, 170 were weak positive, 156 were equivocal and 234 were negative. These results were assessed, and a review of interpretation criteria from both the USA and Europe was carried out. New interpretation criteria and a testing algorithm were developed. The revised criteria changed the results in 109/3688 (3%) patients and produced significantly more Western blot-positive and weak-positive patients than with the current criteria (485 vs. 442, P blot interpretation and improved the sensitivity and robustness of their Western blot method. Using a protocol tailored to patients that incorporates clinical characteristics means that the entire process will be easier and will aid the management of patients.

  12. Clinical, immunohistochemical, Western blot, and genetic analysis in dystrophinopathy.

    Science.gov (United States)

    Na, Sang-Jun; Kim, Won-Joo; Kim, Seung Min; Lee, Kee Ook; Yoon, Bora; Choi, Young-Chul

    2013-08-01

    Dystrophin-deficient muscular dystrophies (dystrophinopathies) are the most common form of muscular dystrophy, with variable clinical phenotypes ranging from the severe Duchenne (DMD) to the milder Becker (BMD) forms. In this study, we investigated the relationship between clinical characteristics, findings at immunohistochemistry (IHC) and Western blot, and the pattern of exon deletions in 24 male patients with dystrophinopathies. We retrospectively reviewed findings from clinical and laboratory examinations, IHC for dystrophin of muscle biopsy tissue, Western blot analysis, and multiplex polymerase chain reaction (PCR) examination of genomic DNA. All tests were performed in every patient. PCR examination revealed exon deletions in 13 patients (54.2%). At Western blot analysis, 15 patients (62.5%) were negative at all three dystrophin domains. Most of these patients had a clinical presentation consistent with the DMD phenotype. Nine (37.5%) others were weakly positive at one or more domains. Most of these patients presented clinically as BMD phenotype. One patient whose clinical presentation was consistent with BMD phenotype had normal findings at IHC and was weakly positive at all three domains on Western blot analysis; however, with the exception of this patient, the findings at IHC and Western blot were consistent for individual patients. Based on these findings, we conclude that Western blot analysis appears useful for confirmation of dystrophinopathy in BMD patients with normal staining on IHC. Exon deletion analysis by multiplex PCR using peripheral blood is also a simple and useful test for the diagnosis of dystrophinopathy, although it has limited sensitivity. Copyright © 2012 Elsevier Ltd. All rights reserved.

  13. 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

  14. A study of the technique of western blot for diagnosis of lyme disease caused by Borrelia afzelii in China.

    Science.gov (United States)

    Liu, Zhi Yun; Hao, Qin; Hou, Xue Xia; Jiang, Yi; Geng, Zhen; Wu, Yi Mou; Wan, Kang Lin

    2013-03-01

    To study the technique of Western blot for the diagnosis of Lyme disease caused by Borrelia afzelii in China and to establish the standard criteria by operational procedure. FP1, which is the representative strain of B. afzelii in China, was analyzed by SDS-PAGE, electro transfer and immunoblotting assays. The molecular weights of the protein bands of FP1 were analyzed by Gel-Pro analysis software. In a study using 451 serum samples (159 patients with Lyme disease and 292 controls), all observed bands were recorded. The accuracy of the WB as a diagnostic test was established by using the ROC curve and Youden index. Criteria for a positive diagnosis of Lyme disease were established as at least one band of P83/100, P58, P39, OspB, OspA, P30, P28, OspC, P17, and P14 in the IgG test and at least one band of P83/100, P58, P39, OspA, P30, P28, OspC, P17, and P41 in the IgM test. For IgG criteria, the sensitivity, specificity and Youden index were 69.8%, 98.3%, and 0.681, respectively; for IgM criteria, the sensitivity, specificity and Youden index were 47%, 94.2%, and 0.412, respectively. Establishment of WB criteria for B. afzelii is important in validating the diagnostic assays for Lyme disease in China. Copyright © 2013 The Editorial Board of Biomedical and Environmental Sciences. Published by China CDC. All rights reserved.

  15. Carbonic anhydrase IX as a specific biomarker for clear cell renal cell carcinoma: comparative study of Western blot and immunohistochemistry and implications for diagnosis.

    Science.gov (United States)

    Giménez-Bachs, José M; Salinas-Sánchez, Antonio S; Serrano-Oviedo, Leticia; Nam-Cha, Syong H; Rubio-Del Campo, Antonio; Sánchez-Prieto, Ricardo

    2012-10-01

    This study aimed to evaluate the usefulness of carbonic anhydrase IX (CA-IX) expression in clear cell renal cell carcinoma (CCRCC) using two different techniques to detect protein expression. An experimental, cross-sectional, analytical study was conducted to analyse proteins in renal tumour and healthy tissue specimens from 38 consecutive patients who underwent nephrectomy for renal cancer. CA-IX protein expression was measured by immunohistochemistry and Western blot analysis and quantified. Statistical analysis was performed with the positive and negative specific agreements and kappa coefficient. The sensitivity and specificity of both techniques were assessed. Statistical tests were conducted to analyse the association between CA-IX expression quantitation and normal prognosis factors (TNM stage and Fuhrman nuclear grade), only in CCRCC. The mean patient age was 65 years, 78.9% of patients were men and 57.9% of tumours were CCRCC. CA-IX protein expression was positive in 63.2% of tumours by immunohistochemistry and in 60.5% by Western blot. Both techniques detected CA-IX expression only in CCRCC and unclassifiable tumours. High concordance indices were observed for CCRCC diagnosis. Western blot and immunohistochemistry had a sensitivity of 95.5% and 100%, respectively; the specificity was 100% in both techniques. CA-IX expression quantitation did not correlate with tumour stage or Fuhrman nuclear grade. Immunochemistry and Western blot techniques can be used to detect abnormal CA-IX protein expression in CCRCC and to support morphology-based diagnostic techniques.

  16. Dealing with large sample sizes: comparison of a new one spot dot blot method to western blot.

    Science.gov (United States)

    Putra, Sulistyo Emantoko Dwi; Tsuprykov, Oleg; Von Websky, Karoline; Ritter, Teresa; Reichetzeder, Christoph; Hocher, Berthold

    2014-01-01

    Western blot is the gold standard method to determine individual protein expression levels. However, western blot is technically difficult to perform in large sample sizes because it is a time consuming and labor intensive process. Dot blot is often used instead when dealing with large sample sizes, but the main disadvantage of the existing dot blot techniques, is the absence of signal normalization to a housekeeping protein. In this study we established a one dot two development signals (ODTDS) dot blot method employing two different signal development systems. The first signal from the protein of interest was detected by horseradish peroxidase (HRP). The second signal, detecting the housekeeping protein, was obtained by using alkaline phosphatase (AP). Inter-assay results variations within ODTDS dot blot and western blot and intra-assay variations between both methods were low (1.04-5.71%) as assessed by coefficient of variation. ODTDS dot blot technique can be used instead of western blot when dealing with large sample sizes without a reduction in results accuracy.

  17. Validation of western blot for Histoplasma capsulatum antibody detection assay.

    Science.gov (United States)

    Almeida, Marcos de Abreu; Pizzini, Cláudia Vera; Damasceno, Lisandra Serra; Muniz, Mauro de Medeiros; Almeida-Paes, Rodrigo; Peralta, Regina Helena Saramago; Peralta, José Mauro; Oliveira, Raquel de Vasconcelos Carvalhaes; Vizzoni, Alexandre Gomes; de Andrade, Carla Lourenço Tavares; Zancopé-Oliveira, Rosely Maria

    2016-02-24

    Histoplasmosis is worldwide systemic mycoses caused by the dimorphic fungus Histoplasma capsulatum. The isolation and identification of H. capsulatum in culture is the reference test for histoplasmosis diagnosis confirmation. However, in the absence of it, serology has been used as a presumptive diagnosis through antibody and antigen detection. The purpose of the present study was to validate an immunoassay method (western blot) for antibodies detection in the diagnosis of histoplasmosis. To validate the western blot (WB) a study was conducted using 118 serum samples from patients with histoplasmosis and 118 serum controls collected from January 2000 to December 2013 in residents of the Rio de Janeiro State, Brazil. Diagnostic validation parameters were calculated based on the categorization of results obtained in a 2 × 2 table and subjected to statistical analysis. In addition, the viability of deglycosylated histoplasmin antigen (ptHMIN) onto nitrocellulose membranes previously sensitized was evaluated during the same period. The WB test showed sensitivity of 94.9 %, specificity of 94.1 %, positive predictive value of 94.1 %, negative predictive value of 94.9 %, accuracy of 94.5 %, and almost perfect precision. Besides, the strips have proved to be viable for using at least 5 years after ptHMIN antigen sensitization. Western blot test using ptHMIN provides sensitive, specific, and faster results. Therefore, could be considered a useful tool in the diagnosis of histoplasmosis being used by public health system, even in situations where laboratory facilities are relatively limited.

  18. A defined methodology for reliable quantification of Western blot data.

    Science.gov (United States)

    Taylor, Sean C; Berkelman, Thomas; Yadav, Geetha; Hammond, Matt

    2013-11-01

    Chemiluminescent western blotting has been in common practice for over three decades, but its use as a quantitative method for measuring the relative expression of the target proteins is still debatable. This is mainly due to the various steps, techniques, reagents, and detection methods that are used to obtain the associated data. In order to have confidence in densitometric data from western blots, researchers should be able to demonstrate statistically significant fold differences in protein expression. This entails a necessary evolution of the procedures, controls, and the analysis methods. We describe a methodology to obtain reliable quantitative data from chemiluminescent western blots using standardization procedures coupled with the updated reagents and detection methods.

  19. 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.

  20. The design of a quantitative western blot experiment.

    Science.gov (United States)

    Taylor, Sean C; Posch, Anton

    2014-01-01

    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.

  1. 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.

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

    Science.gov (United States)

    Takesue, Masataka; Osaka, Yuki; Muranaka, Masanori; Katayama, Yoshinari; Ikadai, Hiromi

    2016-01-01

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

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

    Science.gov (United States)

    Sackstein, Robert; Fuhlbrigge, Robert

    2015-01-01

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

  4. Application of Intermittent Microwave Irradiation to Western Blot Analysis.

    Science.gov (United States)

    Liu, Yu-Ting; Toyokuni, Shinya

    2015-01-01

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

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

    Science.gov (United States)

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

    2014-03-01

    An important concern for the use of antibodies in various applications, such as western blot (WB) or immunohistochemistry (IHC), is specificity. This calls for systematic validations using well-designed conditions. Here, we have analyzed 13 000 antibodies using western blot with lysates from human cell lines, tissues, and plasma. Standardized stratification showed that 45% of the antibodies yielded supportive staining, and the rest either no staining (12%) or protein bands of wrong size (43%). A comparative study of WB and IHC showed that the performance of antibodies is application-specific, although a correlation between no WB staining and weak IHC staining could be seen. To investigate the influence of protein abundance on the apparent specificity of the antibody, new WB analyses were performed for 1369 genes that gave unsupportive WBs in the initial screening using cell lysates with overexpressed full-length proteins. Then, more than 82% of the antibodies yielded a specific band corresponding to the full-length protein. Hence, the vast majority of the antibodies (90%) used in this study specifically recognize the target protein when present at sufficiently high levels. This demonstrates the context- and application-dependence of antibody validation and emphasizes that caution is needed when annotating binding reagents as specific or cross-reactive. WB is one of the most commonly used methods for validation of antibodies. Our data implicate that solely using one platform for antibody validation might give misleading information and therefore at least one additional method should be used to verify the achieved data. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  6. 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.

  7. Metabotropic glutamate receptor 3 (mGlu3; mGluR3; GRM3) in schizophrenia: Antibody characterisation and a semi-quantitative western blot study.

    Science.gov (United States)

    García-Bea, Aintzane; Walker, Mary A; Hyde, Thomas M; Kleinman, Joel E; Harrison, Paul J; Lane, Tracy A

    2016-11-01

    Metabotropic glutamate receptor 3 (mGlu3, mGluR3), encoded by GRM3, is a risk gene for schizophrenia and a therapeutic target. It is unclear whether expression of the receptor is altered in the disorder or related to GRM3 risk genotype. Antibodies used to date to assess mGlu3 in schizophrenia have not been well validated. To characterise six commercially available anti-mGlu3 antibodies for use in human brain, and then conduct a semi-quantitative study of mGlu3 immunoreactivity in schizophrenia. Antibodies tested using Grm3-/- and Grm2-/-/3-/- mice and transfected HEK293T/17 cells. Western blotting on membrane protein isolated from superior temporal cortex of 70 patients with schizophrenia and 87 healthy comparison subjects, genotyped for GRM3 SNP rs10234440. One (out of six) anti-mGlu3 antibodies was fully validated, a C-terminal antibody which detected monomeric (~100kDa) and dimeric (~200kDa) mGlu3. A second, N-terminal, antibody detected the 200kDa band but also produced non-specific bands. Using the C-terminal antibody for western blotting in human brain, mGlu3 immunoreactivity was found to decline with age, and was affected by pH and post mortem interval. There were no differences in monomeric or dimeric mGlu3 immunoreactivity in schizophrenia or in relation to GRM3 genotype. The antibody was not suitable for immunohistochemistry. These data highlight the value of knockout mouse tissue for antibody validation, and the need for careful antibody characterisation. The schizophrenia data show that involvement of GRM3 in the disorder and its genetic risk architecture is not reflected in total membrane mGlu3 immunoreactivity in superior temporal cortex. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2015-05-30

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

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

    Science.gov (United States)

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

    2014-01-01

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

  10. The fastest Western in town: a contemporary twist on the classic Western blot analysis.

    Science.gov (United States)

    Silva, Jillian M; McMahon, Martin

    2014-02-05

    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 conventional Laemmli system, generates better protein separation and resolution, maintains protein integrity, and reduces electrophoresis to a 35 min run time. Moreover, the iBlot dry blotting system, dramatically improves the efficacy and speed of protein transfer to the membrane in 7 min, which is in contrast to the traditional protein transfer methods that are often more inefficient with lengthy transfer times. In combination with these highly innovative modifications, protein detection using infrared fluorescent imaging results in higher-quality, more accurate and consistent data compared to the standard Western blotting technique of chemiluminescence. This technology can simultaneously detect two different antigens on the same membrane by utilizing two-color near-infrared dyes that are visualized in different fluorescent channels. Furthermore, the linearity and broad dynamic range of fluorescent imaging allows for the precise quantification of both strong and weak protein bands. Thus, this protocol describes the key improvements to the classic Western blotting method, in which these advancements significantly increase the quality of data while greatly reducing the performance time of this experiment.

  11. The Multispot rapid HIV-1/HIV-2 differentiation assay is comparable with the Western blot and an immunofluorescence assay at confirming HIV infection in a prospective study in three regions of the United States.

    Science.gov (United States)

    Pandori, Mark W; Westheimer, Emily; Gay, Cindy; Moss, Nicholas; Fu, Jie; Hightow-Weidman, Lisa B; Craw, Jason; Hall, Laura; Giancotti, Francesca R; Mak, Mae Ling; Madayag, Carmela; Tsoi, Benjamin; Louie, Brian; Patel, Pragna; Owen, S Michele; Peters, Philip J

    2013-12-01

    A new HIV diagnostic algorithm has been proposed which replaces the use of the HIV-1 Western blot and HIV-1 immunofluorescence assays (IFA) as the supplemental test with an HIV-1/HIV-2 antibody differentiation assay. To compare an FDA-approved HIV-1/HIV-2 antibody differentiation test (Multispot) as a confirmatory test with the HIV-1 Western blot and IFA. Participants were screened with an HIV-1/HIV-2 combination Antigen/Antibody (Ag/Ab) screening assay. Specimens with repeatedly reactive results were tested with Multispot and either Western blot or IFA. Specimens with discordant screening and confirmatory results were resolved with HIV-1 RNA testing. Individuals (37,876) were screened for HIV infection and 654 (1.7%) had a repeatedly reactive Ag/Ab assay result. On Multispot, 554 (84.7%) were HIV-1 reactive, 0 (0%) were HIV-2 reactive, 1 (0.2%) was reactive for both HIV-1 and HIV-2 (undifferentiated), 9 (1.4%) were HIV-1 indeterminate, and 90 (13.8%) were non-reactive. HIV-1 RNA was detected in 47/90 Multispot non-reactive (52.2%) specimens. Among specimens confirmed to have HIV infection (true positives), Multispot and Western blot detected HIV-1 antibody in a similar proportion of cases (93.7% vs. 94.4% respectively) while Multispot and IFA also detected HIV-1 antibody in a similar proportion of cases (84.5% vs. 83.4% respectively). In this study, Multispot confirmed HIV infections at a similar proportion to Western blot and IFA. Multispot, Western blot, and IFA, however, did not confirm all of the reactive Ag/Ab assay results and underscores the importance of HIV NAT testing to resolve discordant screening and confirmatory results. Copyright © 2013 The Authors. Published by Elsevier B.V. All rights reserved.

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

    Science.gov (United States)

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

    2015-09-01

    Toxoplasma gondii can infect almost all warm-blooded animals, and many serological methods have been developed to detect T. gondii infection in a variety of animal species. In the present study, the seroprevalence of T. gondii infection in farmed mink in northeast China was determined using the modified agglutination test (MAT), a Western blot (WB), and 3 enzyme-linked immunosorbent assays (ELISAs) with protein A/G conjugate, using either of 2 recombinant dense granule antigens, GRA1 and GRA7, or Toxoplasma soluble antigens (TSA). There was no significant difference between the detection results of the GRA1-, GRA7-, and TSA-ELISAs and WB (McNemar chi-square, P > 0.05), but a significant difference was observed between MAT and WB (P < 0.05). A near perfect agreement (97.0%) was found between the GRA7-ELISA and WB (κ = 0.83), and a substantial agreement (92.4-93.1%) was observed in the TSA- and GRA1-ELISAs (κ = 0.68-0.73). The GRA7-ELISA showed the highest sensitivity and specificity, and the lowest false-positive and negative rates, while the MAT gave both a low sensitivity and frequent false positives in comparison to the WB. Receiver operating characteristic analysis revealed the largest area under curve of 0.85 (95% confidence interval: 0.74-0.96), and the highest relative sensitivity (72.7%) and specificity (99.0%) for a cutoff value of 0.19 in the GRA7-ELISA. These results indicate that the GRA7-ELISA is suitable for detection of T. gondii infection in mink and that MAT should be used with caution. © 2015 The Author(s).

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

    Science.gov (United States)

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

    2012-06-15

    Comparing protein levels from single cells in tissue has not been achieved through Western blot. Laser capture microdissection allows for the ability to excise single cells from sectioned tissue and compile an aggregate of cells in lysis buffer. In this study we analyzed proteins from cells excised individually from brain and muscle tissue through Western blot. After we excised individual neurons from the substantia nigra of the brain, the accumulated surface area of the individual cells was 120,000, 24,000, 360,000, 480,000, 600,000 μm2. We used an optimized Western blot protocol to probe for tyrosine hydroxylase in this cell pool. We also took 360,000 μm2 of astrocytes (1700 cells) and analyzed the specificity of the method. In muscle we were able to analyze the proteins of the five complexes of the electron transport chain through Western blot from 200 human cells. With this method, we demonstrate the ability to compare cell-specific protein levels in the brain and muscle and describe for the first time how to visualize proteins through Western blot from cells captured individually. Copyright © 2012 Elsevier Inc. All rights reserved.

  14. 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.

  15. Serological follow-up of patients with paracoccidioidomycosis treated with itraconazole using Dot-blot, ELISA and Western-blot

    Directory of Open Access Journals (Sweden)

    Rosana MARTINS

    1997-09-01

    Full Text Available Twenty-seven mycologically proven cases of paracoccidioidomycosis (PCM were treated with itraconazole (100-200 mg/day in month 1 and 100 mg/day until month 6-8 and evaluated clinically and serologically, up to 3.5 years post-therapy, using Dot-blot and ELISA for measuring the titers of IgG, IgA and IgM anti-P. brasiliensis antibodies and Western-blot for determining IgG, IgA and IgM antibodies against the antigen components of the fungus. Before treatment, 81.5% (Dot-blot and 84% (ELISA of the patients presented elevated IgG anti-P. brasiliensis antibody titers which dropped slightly with treatment. On the other hand, the percentages of pre-treatment high-titered sera for IgA and IgM anti-P.brasiliensis were lower (5l.9% and 5l.8%: Dot-blot; 16.5 and 36%: ELISA, respectively but the titers tended to become negative more frequently with treatment. Prior to treatment, the percentages of positivity for IgG, IgA and IgM anti-P.brasiliensis antibodies in Western-blot were 96%, 20.8% and 41.6%, respectively. Antigens with molecular weights varying from 16-78 kDa, from 21-76 kDa and from 27-78 kDa were reactive for IgG, IgA and IgM antibodies, respectively. The most frequently reactive antigenic components had molecular weights of 27, 33 and 43 kDa for IgG, and 70 for IgA and IgM antibodies. During the period of study, the patients responded well to treatment. The present data confirm the diversity and complexity of the humoral response in PCM, and the importance of utilizing different serological tests to detect IgG, IgA and IgM anti-P. brasiliensis antibodiesVinte e sete pacientes portadores de paracoccidioidomicose (PCM foram tratados com itraconazole (100-200 mg/dia no primeiro mês e 100 mg/dia até 6-8 meses e avaliados sob o ponto de vista clínico e sorológico, até 3 e meio anos após o início do tratamento, utilizando-se os testes de Dot-blot e ELISA para medir os títulos de anticorpos IgG, IgA e IgM anti-P. brasiliensis, e Western-blot

  16. Evaluation of an adaptive virtual laboratory environment using Western Blotting for diagnosis of disease.

    Science.gov (United States)

    Polly, Patsie; Marcus, Nadine; Maguire, Danni; Belinson, Zack; Velan, Gary M

    2014-10-20

    Providing large numbers of undergraduate students in scientific disciplines with engaging, authentic laboratory experiences is important, but challenging. Virtual laboratories (vLABs) are a potential means to enable interactive learning experiences. A vLAB focusing on Western Blotting was developed and implemented in a 3rd year undergraduate Pathology course for science students to facilitate learning of technical molecular laboratory skills that are linked to development of diagnostic skills. Such skills are important for undergraduates in building a conceptual understanding of translation of laboratory techniques to changes in human biology due to disease. The Western Blotting vLAB was developed and deployed using the Adaptive eLearning Platform (AeLP) developed by Smart Sparrow (https://www.smartsparrow.com/). The vLAB was evaluated to assess students' perceptions of their laboratory skills relevant to the diagnosis of Muscular Dystrophy. A blended learning rotation model was applied in which wet laboratory and vLAB environments for Western Blotting were both delivered to three consecutive cohorts of 3rd year science undergraduates undertaking a Muscle Diseases practical class. Evaluation questionnaires were administered at the completion of the practical classes. Students indicated in online questionnaires that the Western Blotting vLAB was at least equivalent to the real lab in their perceived development of concepts, laboratory skills and diagnosis of disease. vLABs have great potential for improving students' development of diagnostic skills. Further studies are required to determine the impact of vLABs on student learning.

  17. Blame it on Southern, but it's a western blot.

    Science.gov (United States)

    Klionsky, Daniel J

    2017-01-02

    Edwin M. Southern is a professor emeritus at the University of Oxford. He is perhaps best known for development of the "Southern blot" (Dr. Southern was at the University of Edinburgh when he wrote his landmark paper). The Southern blot provided a scientific breakthrough by allowing scientists to detect a particular DNA sequence without first purifying it from the rest of the genome; the basic method involves the transfer of the DNA to a membrane, followed by detection with a specific probe. Although few people perform Southern blots as originally carried out by Southern, due in part to the more recent technique of the polymerase chain reaction, the basic concept continues to play an important role in molecular biology.

  18. Native Electrophoresis and Western Blot Analysis (NEWeB): Methods and Applications.

    Science.gov (United States)

    Manoussopoulos, Ioannis N; Tsagris, Mina

    2015-01-01

    Native Electrophoresis and Western Blot Analysis (NEWeB) has been developed for the study of plant virus characteristics, among others, virus particle-protein interactions, electrophorotype formation, and strain separation. The method is based on the property of electrophoretic mobility of virus particles (VP) and proteins and combines the analytical capacity of electrophoresis with the specificity of western blot. One of its advantages is that it deals with entire VP that can be studied in cause and effect or in time-interval experiments. Some of the most interesting approaches include VP structural studies, VP interaction with host or viral proteins, and also the characterization of VP-protein complexes. In this protocol, NEWeB is used to demonstrate the interaction of Plum pox virus particles with the helper component, a virus encoded protein. It is expected that the method could be used in analogous studies of other viruses or large protein complexes, where similar principles apply.

  19. 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

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

    Science.gov (United States)

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

    2016-01-01

    Currently, Western blot is used to confirm the initial serodiagnosis of HIV infection by antibody detection. However, a major deficiency of the Western blot relates to a lack of sufficient sensitivity in detecting HIV antibodies. This report describes a simple, sensitive and inexpensive bead-based assay for detection of early HIV infection. A panel of 138 positive specimens including 105 blood donors and 33 MSM with known Western blot results were evaluated using Luminex xMAP at Tianjin Centers for Disease Control and Prevention (CDC). We demonstrate a superior sensitivity of Luminex xMAP compared with Western blot. Of the 87 confirmed HIV positive blood donors, Western blot only confirmed 65 cases with 74.7% (65/87) sensitivity while Luminex xMAP identified 72 cases with 82.8% (72/87) sensitivity (pWestern blot and Luminex xMAP verified 13 and 19 of 33 MSM specimens, respectively. The sensitivity was 39.4% (13/33) for Western blot and 57.6% (19/33) for Luminex xMAP (pWestern blot improves the diagnostic sensitivity of HIV infection at an early stage, and reduces the chances of missed diagnosis. Copyright © 2015 Elsevier B.V. All rights reserved.

  1. 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…

  2. When less is more: a simple Western blotting amendment allowing data acquisition on human single fibers

    DEFF Research Database (Denmark)

    Jensen, Thomas Elbenhardt; Richter, Erik

    2011-01-01

    This editorial discusses a simple western blotting-amendment allowing rapid data-acquisition on single fibers obtained from freeze-dried human skeletal muscle biopsies.......This editorial discusses a simple western blotting-amendment allowing rapid data-acquisition on single fibers obtained from freeze-dried human skeletal muscle biopsies....

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

    Science.gov (United States)

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

    2015-06-01

    Simple Western is a new technology that allows for the separation, blotting, and detection of proteins similar to a traditional Western except in a capillary format. Traditionally, identity assays for biological products are performed using either an enzyme-linked immunosorbent assay (ELISA) or a manual dot blot Western. Both techniques are usually very tedious, labor-intensive, and complicated for multivalent vaccines, and they can be difficult to transfer to other laboratories. An advantage this capillary Western technique has over the traditional manual dot blot Western method is the speed and the automation of electrophoresis separation, blotting, and detection steps performed in 96 capillaries. This article describes details of the development of an automated identity assay for a 15-valent pneumococcal conjugate vaccine, PCV15-CRM197, using capillary Western technology. Copyright © 2015 Elsevier Inc. All rights reserved.

  4. An appropriate loading control for western blot analysis in animal models of myocardial ischemic infarction.

    Science.gov (United States)

    Nie, Xin; Li, Chen; Hu, Sheng; Xue, Fulai; Kang, Y James; Zhang, Wenjing

    2017-12-01

    An appropriate loading control is critical for Western blot analysis. Housekeeping proteins (HKPs), such as β-actin, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and β-tubulin, are commonly used to normalize protein expression. But HKP expression can be impacted by certain experimental conditions, such as ischemic myocardial infarction. This study was undertaken to look for an appropriate loading control for western blot analysis of ischemic myocardium. Myocardial ischemic infarction was induced by left anterior descending coronary artery (LAD) ligation in Rhesus monkeys and C57BL/6 mice. The heart tissue samples from different areas and time points after surgery were subjected to western blot or gel staining. The level of β-actin, GAPDH, β-tubulin, and total protein were tested. The total protein level was consistent in all groups, whereas the protein level of β-tubulin and β-actin were different in all groups. However, the protein level of GAPDH was stable in the Rhesus monkey model. We concluded that total protein was the most appropriate internal control in different stages of myocardial ischemic disease of various animal models. GAPDH is a reliable internal control only for ischemic myocardium of Rhesus monkey.

  5. β-actin as a loading control for plasma-based Western blot analysis of major depressive disorder patients.

    Science.gov (United States)

    Zhang, Rufang; Yang, Deyu; Zhou, Chanjuan; Cheng, Ke; Liu, Zhao; Chen, Liang; Fang, Liang; Xie, Peng

    2012-08-15

    Western blot analysis is a commonly used technique for determining specific protein levels in clinical samples. For normalization of protein levels in Western blot, a suitable loading control is required. On account of its relatively high and constant expression, β-actin has been widely employed in Western blot of cell cultures and tissue extracts. However, β-actin's presence in human plasma and this protein's putative role as a plasma-based loading control for Western blot analysis remain unknown. In this study, an enzyme-linked immunosorbent assay was used to determine the concentration of β-actin in human plasma, which is 6.29±0.54 ng/ml. In addition, the linearity of β-actin immunostaining and loaded protein amount was evaluated by Western blot, and a fine linearity (R²=0.974±0.012) was observed. Furthermore, the expression of plasma β-actin in major depressive disorder subjects and healthy controls was compared. The data revealed no statistically significant difference between these two groups. Moreover, the total coefficient of variation for β-actin expression in the two groups was 9.2±1.2%. These findings demonstrate that β-actin is present in human plasma and may possibly be used as a suitable loading control for plasma-based Western blot analysis in major depressive disorder. Copyright © 2012 Elsevier Inc. All rights reserved.

  6. The diagnostic value of Western blot method in patients with cystic echinococcosis.

    Science.gov (United States)

    Aslan, Mustafa; Yüksel, Pelin; Polat, Erdal; Cakan, Huseyin; Ergin, Sevgi; Öner, Y Ali; Zengin, Kagan; Arıkan, Soykan; Saribas, Suat; Torun, Muzeyyen Mamal; Kocazeybek, Bekir

    2011-04-01

    Cystic echinococcosis (CE) is the larval cystic stage (called echinococcal cysts) of a small taeniid-type tapeworm (Echinococcus granulosus). Carnivores such as dogs are usually definitive hosts. Intermediate hosts are typically herbivores such as sheep and cattle. CE can be detected using various imaging techniques such as ultrasonography or radiology. Moreover the primary diagnosis has to be confirmed by serological tests since the clinical signs of the disease are non-specific. This study examined the antigenic band patterns useful for serologic diagnosis of hydatidosis. We also report on the post-operative evolution of patients treated for this disease and also determined the diagnostic performance of Western blot IgG kit. Twenty-five (16 females and 9 males) non-operated patients with hydatid cysts (NOP) and 33 (21 females and 12 males) operated patients with hydatid cysts (OP) were included as study group and 22 healthy individuals (14 females and 8 males) with no known chronic diseases were included as a control group. The ages of the patients and control group individuals were between 16-83 years. Patient and control groups were matched for age and sex. Cyst hydatid IgG antibodies were detected in the sera from all patient groups but no antibodies were found in the sera from the control group using ELISA IgG method. Twenty-three (92%) non-operated patients and 18 (54.5%) operated patients exhibited positive results when Western blot IgG kit was used. The P7 band pattern was detected in the sera from all operated and non-operated patients. Twenty-seven of these positive cases had p7 and (p7+p16/18), (p7+p24/26) or (p7+p16/18+p24/26). No antibodies against p7, p16/18 ve p24/26 band patterns were seen in sera from the control group A statistically significant difference was detected between operated and nonoperated patients for Western blot positivity.(pWestern blot kit for 25 cases with CE and 22 healthy controls were calculated as 92%, 100%, 100% and 91

  7. Western blot as a seroepidemiologic tool for detecting foci of Mediterranean spotted fever (MSF).

    Science.gov (United States)

    Raoult, D; Arzouni, J P; Jambon, M C; Beytout, J; Ramousse, O

    1994-02-01

    We conducted a serosurvey on Mediterranean spotted fever (MSF), in a nonendemic area using western blot and microimmunofluorescence. Among 262 tested sera, 53 were positive by micro-immunofluorescence at a titer of 50. When 48 positive sera were western blot tested, 15 did not exhibited any reaction, 17 reacted against the non-specific lipopolysaccharide, and only 16 reacted against the specific protein antigens. Fourteen of the sera with a specific reaction were sampled in a village with a unique submediterranean climate. Western blot may be a more specific tool to determine the real seroprevalence of MSF.

  8. Immunochemische detectiemethoden na western blotting van cytochroom P-450 iso-enzymen

    NARCIS (Netherlands)

    Laan CA; Jansen EHJM

    1992-01-01

    In this report a number of staining techniques on Western blots have been compared with respect to sensitivity, background staining, practical applicability and cost aspects. After electrophoresis of a rat microsomal liver sample followed by blotting, an incubation was performed of a primary

  9. Comparisons of ELISA and Western blot assays for detection of autophagy flux.

    Science.gov (United States)

    Oh, Sung-Hee; Choi, Yong-Bok; Kim, June-Hyun; Weihl, Conrad C; Ju, Jeong-Sun

    2017-08-01

    We analyzed autophagy/mitophagy flux in vitro (C2C12 myotubes) and in vivo (mouse skeletal muscle) following the treatments of autophagy inducers (starvation, rapamycin) and a mitophagy inducer (carbonyl cyanide m-chlorophenylhydrazone, CCCP) using two immunodetection methods, ELISA and Western blotting, and compared their working range, accuracy, and reliability. The ELISAs showed a broader working range than that of the LC3 Western blots (Table 1). Table 2 showed that data value distribution was tighter and the average standard error from the ELISA was much smaller than those of the Western blot, directly relating to the accuracy of the assay. Test-retest reliability analysis showed good reliability for three individual ELISAs (interclass correlation, ≥ 0.7), but poor reliability for three individual Western blots (interclass correlation, ≤ 0.4) (Table 3).

  10. A duplex approach for immunochemical staining and typing of protein in western blots

    NARCIS (Netherlands)

    Kuczius, T.; Brandstädter, L.; Karch, H.; Langeveld, J.P.M.

    2011-01-01

    The qualitative and semiquantitative Western blotting technique enables the detection of separate proteins and the determination of subtypes and fragments by specific immunological reactions. Protein typing on immunoblots is restricted to antibody-specific determination, with the result of a

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

    DEFF Research Database (Denmark)

    Uhlén, Mathias

    2017-01-01

    and then heated to very high temperatures (normally >100 °C) in a procedure that is sometimes termed 'epitope retrieval'. Obviously, this procedure might influence the target protein differently than the procedure used to prepare proteins for a western blot, in which the sample is instead treated with a detergent...... (SDS) before the electrophoresis step. Thus, as concluded by the members of the IWGAV1, the results obtained for a given antibody in western blot applications cannot be used to predict the specificity of the antibody in another assay based on an entirely different epitope-retrieval method, such as IHC...... for specificity in IHC or immunofluorescence microscopy (IF) but which do not stain the correctly sized band in a western blot, and vice versa3, 4, 5. However, as pointed out by Lund-Johansen and Browning, western blot analysis has indeed been found to be useful as a general validation tool for antibody...

  12. Development and evaluation of a Western blot kit for diagnosis of schistosomiasis

    NARCIS (Netherlands)

    Sulahian, Annie; Garin, Yves Jean François; Izri, Arezki; Verret, Caroline; Delaunay, Pascal; van Gool, Tom; Derouin, Francis

    2005-01-01

    We evaluated the performance of Western blot (WB) analysis using commercially available antigen strips and compared the results with those of indirect hemagglutination (IHA) and indirect immunofluorescence (IFAT) for the serodiagnosis of human schistosomiasis. The antigen preparation was a crude

  13. Total Protein Analysis as a Reliable Loading Control for Quantitative Fluorescent Western Blotting

    OpenAIRE

    Eaton, Samantha L.; Roche, Sarah L.; Maica Llavero Hurtado; Oldknow, Karla J.; Colin Farquharson; Gillingwater, Thomas H.; Wishart, Thomas M

    2013-01-01

    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 ...

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

    Science.gov (United States)

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

    2012-03-01

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

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

    Science.gov (United States)

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

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

  16. 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.

  17. SDS-PAGE and Western blot of urinary proteins in dogs with leishmaniasis.

    Science.gov (United States)

    Zaragoza, Concepción; Barrera, Rafael; Centeno, Francisco; Tapia, Jose A; Durán, Esther; González, Marta; Mañé, M Cinta

    2003-01-01

    Canine leishmaniasis is an endemic disease in the Mediterranean area caused by the protozoan Leishmania infantum, which usually produces renal failure. Sodium dodecyl sulphate polyacrylamide gel electrophoresis and Western blot using antibodies to IgG and IgA from dogs were carried out in the urine of 22 dogs with leishmaniasis diagnosed by ELISA and confirmed by PCR, and 20 healthy dogs. The results were compared to renal function laboratory tests and to those from a histopathological study of the kidneys from sick animals that died naturally or were euthanized. Five different bands with molecular weights ranging from 10 to 110 kDa were obtained from the electrophoresis of the urine of healthy dogs. 33.5% of total proteins corresponded to low molecular weight proteins and the other proteins had middle and high molecular weights. However, in the group with leishmaniasis, a maximum of 11 different bands with molecular weights ranging from 10 kDa to 150 kDa were displayed in the electrophoresis of the urine. The urine electrophoretic pattern in the sick dogs was classified as mixed (proteins with high and low molecular weights) because low molecular weight proteins made up 57.9% and the rest of the proteins had middle and high molecular weights. In Western blot, none of the healthy dogs showed excretion of IgG and/or IgA, whereas IgG and IgA were detected in the Western blot of urine of 68% and 55% respectively of dogs with leishmaniasis. The results obtained in the leishmaniasis group agreed with glomerular and tubular damage, which were confirmed by the histopathological findings.

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

    OpenAIRE

    Meshgi, B; A Eslami; Halajian, A.

    2009-01-01

    "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 ...

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

    Science.gov (United States)

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

    2000-01-01

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

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

    Science.gov (United States)

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

    2015-01-01

    Antibodies are important tools for the study of protein expression but are often used without full validation. In this study, we used Western blots to characterize antibodies targeted to the N or C terminal (NT or CT, respectively) and the second or third intracellular loop (IL2 or IL3, respectively) of the endothelin B receptor (ETB). The IL2-targeted antibody accurately detected endogenous ETB expression in rat brain and cultured rat astrocytes by labeling a 50-kDa band, the expected weight of full-length ETB. However, this antibody failed to detect transfected ETB in HEK293 cultures. In contrast, the NT-targeted antibody accurately detected endogenous ETB in rat astrocyte cultures and transfected ETB in HEK293 cultures by labeling a 37-kDa band but failed to detect endogenous ETB in rat brain. Bands detected by the CT- or IL3-targeted antibody were found to be unrelated to ETB. Our findings show that functional ETB can be detected at 50 or 37kDa on Western blot, with drastic differences in antibody affinity for these bands. The 37-kDa band likely reflects ETB processing, which appears to be dependent on cell type and/or culture condition. Copyright © 2014 Elsevier Inc. All rights reserved.

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

    Science.gov (United States)

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

    2014-11-20

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

  2. Enrichment of PrPSc in formalin-fixed, paraffin-embedded tissues prior to analysis by Western blot.

    Science.gov (United States)

    Nicholson, Eric M

    2011-07-01

    Diagnosis of prion disease is primarily through immunodetection of the infectious agent. Typically, 2 distinct procedures are recommended for a definitive diagnosis, with immunohistochemistry and Western blot providing the most information as to the specific isolate in question. In the past, these approaches required formalin-fixed, paraffin-embedded tissue and fresh or frozen tissue, respectively; however, methods have been developed that allow for use of fixed tissue for Western blot. The present study describes a method of enriching PrP(Sc) in formalin-fixed, paraffin-embedded tissues prior to Western blot analysis for the detection of PrP(Sc). With this modified procedure, 5 times the previously reported sample size may be used for analysis, greatly enhancing the sensitivity of this procedure.

  3. Construction of multiple-epitope tag sequence by PCR for sensitive Western blot analysis.

    Science.gov (United States)

    Nakajima, K; Yaoita, Y

    1997-01-01

    Epitope tagging is a powerful technique to characterize a recombinantly expressed protein encoded by cDNA without the purification of the protein and the immunization of animals. In some cases, however, the expression of a tagged protein is too low to analyze by Western blot. We have developed a simple method to generate tandem repetitive nucleotide sequence by PCR, which allows us to label a protein of interest with a multiple-epitope tag. When five myc epitopes were attached to vaccinia virus protein CrmA, its signal was multiplied 5.8 times in Western blot analysis, compared with that of one epitope-tagged CrmA. PMID:9153327

  4. Western blot analysis of cells encapsulated in self-assembling peptide hydrogels.

    Science.gov (United States)

    Burgess, Kyle A; Miller, Aline F; Oceandy, Delvac; Saiani, Alberto

    2017-12-01

    Continuous optimization of in vitro analytical techniques is ever more important, especially given the development of new materials for tissue engineering studies. In particular, isolation of cellular components for downstream applications is often hindered by the presence of biomaterials, presenting a major obstacle in understanding how cell-matrix interactions influence cell behavior. Here, we describe an approach for western blot analysis of cells that have been encapsulated in self-assembling peptide hydrogels (SAPHs), which highlights the need for complete solubilization of the hydrogel construct. We demonstrate that both the choice of buffer and multiple cycles of sonication are vital in obtaining complete solubilization, thereby enabling the detection of proteins otherwise lost to SAP aggregation. Moreover, we show that the presence of self-assembling peptides (SAPs) does not interfere with the standard immunoblotting technique, offering the potential for use in more full-scale proteomic studies.

  5. 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…

  6. 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...

  7. Two-dimensional gel-based protein standardization verified by western blot analysis.

    Science.gov (United States)

    Haniu, Hisao; Watanabe, Daisuke; Kawashima, Yusuke; Matsumoto, Hiroyuki

    2015-01-01

    In data presentation of biochemical investigation the amount of a target protein is shown in the y-axis against the x-axis representing time, concentrations of various agents, or other parameters. Western blot is a versatile and convenient tool in such an analysis to quantify and display the amount of proteins. In western blot, so-called housekeeping gene product(s), or "housekeeping proteins," are widely used as internal standards. The rationale of using housekeeping proteins for standardization of western blot is based on the assumption that the expression of chosen housekeeping gene is always constant, which could be false under certain physiological or pathological conditions. We have devised a two-dimensional gel-based standardization method in which the protein content of each sample is determined by scanning the total protein density of two-dimensional gels and the expression of each protein is quantified as the density ratio of each protein divided by the density of the total proteins on the two-dimensional gel. The advantage of this standardization method is that it is not based on any presumed "housekeeping proteins" that are supposed to be being expressed constantly under all physiological conditions. We will show that the total density of a two-dimensional gel can render a reliable protein standardization parameter by running western blot analysis on one of the proteins analyzed by two-dimensional gels.

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

    Science.gov (United States)

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

  9. A western blot protocol for detection of proteins heterologously expressed in Xenopus laevis oocytes

    DEFF Research Database (Denmark)

    Jørgensen, Morten Egevang; Nour-Eldin, Hussam Hassan; Halkier, Barbara Ann

    2016-01-01

    at the individual oocyte level is often desirable when comparing properties of wild type and mutant transporters. However, a large content of yolk platelets in the oocyte cytoplasm makes this a challenging task. Here we report a method for fast and easy, semiquantitative Western blot analysis of proteins...

  10. Indeterminate human immunodeficiency virus Western blot profiles in ethiopians with discordant screening-assay results

    NARCIS (Netherlands)

    Meles, Hailu; Wolday, Dawit; Fontanet, Arnaud; Tsegaye, Aster; Tilahun, Tesfaye; Aklilu, Mathias; Sanders, Eduard; Rinke de Wit, Tobias F.

    2002-01-01

    The Western blot (WB) assay is the most widely accepted confirmatory assay for the detection of antibodies to human immunodeficiency virus type 1 (HIV-1). However, indeterminate WB reactivity to HIV-1 proteins may occur in individuals who do not appear to be infected with HIV. The profiles of WB

  11. Western Blot Detection of Human Anti-Chikungunya Virus Antibody with Recombinant Envelope 2 Protein.

    Science.gov (United States)

    Yang, Zhaoshou; Lee, Jihoo; Ahn, Hye-Jin; Chong, Chom-Kyu; Dias, Ronaldo F; Nam, Ho-Woo

    2016-04-01

    Chikungunya virus (CHIKV), a tropical pathogen, has re-emerged and has massive outbreaks abruptly all over the world. Containing many dominant epitopes, the envelope E2 protein of CHIKV has been explored for the vaccination or diagnosis. In the present study, the antigenicity of a recombinant expressed intrinsically disorder domain (IUD) of E2 was tested for the detection of the antibody against CHIKV through western blot method. The gene of the IUD of E2 was inserted into 2 different vectors and expressed as recombinant GST-E2 and recombinant MBP-E2 fusion protein, respectively. Two kinds of fusion proteins were tested with 30 CHIKV patient sera and 30 normal sera, respectively. Both proteins were detected by 25 patients sera (83.3%) and 1 normal serum (3.3%). This test showed a relatively high sensitivity and very high specificity of the recombinant E2 proteins to be used as diagnostic antigens against CHIKV infection.

  12. 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.

  13. Characterization of somatic antigens of adult Toxocara canis by western blotting

    Directory of Open Access Journals (Sweden)

    Shivani Sahu

    Full Text Available Aim: The objective of this study was characterize the somatic soluble antigens of adult Toxocara canis (Tc-SA by western blotting. Materials and Methods: T. canis worms were collected from the naturally infected pups after deworming. The somatic antigen was prepared as per standard procedure with slight modification. These antigens were separated using Sodium dodecyl sulphate-electrophoresis (SDS-PAGE. The specific reactivity of the Tc-SA proteins was checked against the serum of naturally infected dogs as well with the hyperimmune serum raised in the rabbit by western blotting. Results: On SDS-PAGE recovered proteins ranged in size from 44 to 300 kDa. The immuno-reactivity of the naturally infected dog sera with the Tc-SA antigens showed 12 prominent immunoreactive bands of distinct sizes at 28.61, 32.60, 38.10, 43.04, 49.99, 67.57, 73.22, 105.77, 144.74, 161.11, 177.84 and 196.31 kDa. The immuno-reactivity of the hyper immune serum raised in rabbits against Tc-SA antigens was observed with 10 prominent bands of distinct sizes at 17.11, 24.15, 34.83, 43.46, 52.47, 55.89, 67.57, 70, 74.60 and 105.6 kDa. Conclusions: Common antigens band were observed at 67 and 105 kDa. These antigens merit further evaluation as candidate for use in diagnosis of toxocarosis in humans and adult dogs. [Vet World 2013; 6(7.000: 424-427

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

    Science.gov (United States)

    Xu, Dong; Mane, Sarthak; Sosic, Zoran

    2015-01-01

    This paper describes the application of an automated size-based capillary Western blot system (Sally instrument) from ProteinSimple, Inc., for biopharmaceutical fusion-Fc protein characterization and evaluation of its purification process. The fusion-Fc protein column purification from an excess of single chain Fc polypeptide and removal of an enzyme coexpressed for protein maturation have been demonstrated using an automated capillary Western system. The clearance of a selected host cell protein (HCP) present in cell culture of fusion-Fc protein was also quantitatively monitored throughout the protein purification process. Additionally, the low levels of fusion-Fc product-related impurities detected by traditional slab gel Western blot were confirmed by the automated capillary Western system. Compared to the manual approach, the automated capillary Western blot provides the advantages of ease of operation, higher sample throughput, greater linearity range, and higher precision for protein quantitation. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. Better management of Western blotting results using professional photo management software.

    Science.gov (United States)

    Iorio-Morin, Christian; Germain, Pascale; Parent, Jean-Luc

    2013-04-01

    Western blotting is a proven technique essential to a significant proportion of molecular biology projects. However, as results accumulate over the years, managing data can become daunting. Recognizing that the needs of a scientist working with Western blotting results are conceptually the same as those of a professional photographer managing a summer's worth of wedding photos, we report here a new workflow for managing Western blotting results using professional photo management software. The workflow involves (i) scanning all film-based results; (ii) importing the scans into the software; (iii) processing the scans; (iv) tagging the files with metadata, and (v) creating appropriate "smart-albums." Advantages of this system include space savings (both on our hard drives and on our desks), safer archival, quicker access, and easier sharing of the results. In addition, metadata-based workflows improve cross-experiment discovery and enable questions like "show me all blots labelled with antibody X" or "show me all experiments featuring protein Y". As project size and breadth increase, workflows delegating results management to the computer will become more and more important so that scientists can keep focussing on science. © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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

    Science.gov (United States)

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

    2016-01-01

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

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

    Science.gov (United States)

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

    2016-05-01

    Serology has a pivotal role in the diagnosis of congenital syphilis (CS), but problems arise because of the passive transfer of IgG antibodies across the placenta. The aim of this study was to assess the diagnostic value of a comparative Western blot (WB) method finalized to match the IgG immunological profiles of mothers and their own babies at birth in order to differentiate between passively transmitted maternal antibodies and antibodies synthesized by the infants against Treponema pallidum Thirty infants born to mothers with unknown or inadequate treatment for syphilis were entered in a retrospective study, conducted at St. Orsola-Malpighi Hospital, Bologna, Italy. All of the infants underwent clinical, instrumental, and laboratory examinations, including IgM WB testing. For the retrospective study, an IgG WB assay was performed by blotting T. pallidum antigens onto nitrocellulose sheets and incubating the strips with serum specimens from mother-child pairs. CS was diagnosed in 11 out of the 30 enrolled infants; 9/11 cases received the definitive diagnosis within the first week of life, whereas the remaining two were diagnosed later because of increasing serological test titers. The use of the comparative IgG WB testing performed with serum samples from mother-child pairs allowed a correct CS diagnosis in 10/11 cases. The CS diagnosis was improved by a strategy combining comparative IgG WB results with IgM WB results, leading to a sensitivity of 100%. The comparative IgG WB test is thus a welcome addition to the conventional laboratory methods used for CS diagnosis, allowing identification and adequate treatment of infected infants and avoiding unnecessary therapy of uninfected newborns. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

  18. 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.

  19. 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.

  20. Imported intraocular gnathostomiasis with subretinal tracks confirmed by western blot assay.

    Science.gov (United States)

    Yang, Ji Ho; Kim, Moosang; Kim, Eung Suk; Na, Byoung-Kuk; Yu, Seung-Young; Kwak, Hyung-Woo

    2012-03-01

    We report a case of intraocular gnathostomiasis diagnosed by western blot assay in a patient with subretinal tracks. A 15-year-old male patient complained of blurred vision in the right eye, lasting for 2 weeks. Eight months earlier, he had traveled to Vietnam for 1 week and ate raw wild boar meat and lobster. His best-corrected visual acuity was 20/20 in both eyes and anterior chamber examination revealed no abnormalities. Fundus examination showed subretinal tracks in the right eye. Fluorescein angiography and indocyanine green angiography showed linear hyperfluorescence of the subretinal lesion observed on fundus in the right eye. Ultrasound examination revealed no abnormalities. Blood tests indicated mild eosinophilia (7.5%), and there was no abnormality found by systemic examinations. Two years later, the patient visited our department again for ophthalmologic evaluation. Visual acuity remained 20/20 in both eyes and the subretinal tracks in the right eye had not changed since the previous examination. Serologic examination was performed to provide a more accurate diagnosis, and the patient's serum reacted strongly to the Gnathostoma nipponicum antigen by western blot assay, which led to a diagnosis of intraocular gnathostomiasis. This is the first reported case of intraocular gnathostomiasis with subretinal tracks confirmed serologically using western blot in Korea.

  1. 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.

  2. More specific bands in the IgG western blot in sera from Scottish patients with suspected Lyme borreliosis.

    Science.gov (United States)

    Evans, Roger; Mavin, Sally; McDonagh, Susan; Chatterton, Jean M W; Milner, Rachel; Ho-Yen, Darrel O

    2010-08-01

    To identify further Western blot bands that may be specific in the diagnosis of Lyme borreliosis. The Borrelia burgdorferi antibody profiles of 270 western blot positive patients and 241 western blot negative patients from 2008 were examined. 27 different non-specific bands were detected in both groups. Six of 27 (22%) of the non-specific bands were detected significantly more in the western blot positive patients compared to the western blot negative patients (20 kDa, p<0.0001; 28 kDa, p<0.002; 36 kDa, p<0.002; 37 kDa, p<0.007; 48 kDa, p<0.023; 56 kDa, p<0.028; two-tailed F test). Results suggest that the 20, 28 and 48 kDa bands should be regarded as specific.

  3. 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.

  4. Detection of early antibodies in human immunodeficiency virus infection by enzyme-linked immunosorbent assay, Western blot, and radioimmunoprecipitation.

    OpenAIRE

    Saah, A J; Farzadegan, H; Fox, R; Nishanian, P; Rinaldo, C R; Phair, J P; Fahey, J L; Lee, T H; Polk, B F

    1987-01-01

    A current concept of the serological response to human immunodeficiency virus (HIV) infection in humans is that antibodies to core antigens (p55, p24, and p15) are detectable earlier during initial stages of antibody production than antibodies against envelope antigens (gp160, gp120, and gp41). Comparative studies of Western blot (immunoblot), radioimmunoprecipitation assay (RIPA), and enzyme-linked immunosorbent assay (ELISA) during initial antibody production are limited to case reports and...

  5. Herpes simplex virus type 2 antibody detection performance in Kisumu, Kenya, using the Herpeselect ELISA, Kalon ELISA, Western blot and inhibition testing.

    Science.gov (United States)

    Smith, J S; Bailey, R C; Westreich, D J; Maclean, I; Agot, K; Ndinya-Achola, J O; Hogrefe, W; Morrow, R A; Moses, S

    2009-04-01

    In certain parts of Africa, type-specific herpes simplex virus type 2 (HSV-2) ELISAs may have limited specificity. To date, no study has been conducted to validate HerpeSelect and Kalon type-specific HSV-2 ELISAs using both the Western blot and recombinant gG ELISA inhibition testing as reference standards. A total of 120 men who were HIV seronegative (aged 18-24 years) provided blood samples. HSV-2 IgG serum antibodies were detected using four different methods: HerpeSelect HSV-2 ELISA (n = 120), Kalon HSV-2 ELISA (n = 120), University of Washington Western blot (n = 101) and a recombinant inhibition test (n = 93). HSV-2 seroprevalence differed significantly by HSV-2 detection method, ranging from 24.8% with the Western blot to 69.8% with the HerpeSelect ELISA. Using the Western blot as the reference standard, the HerpesSelect had the highest sensitivity for HSV-2 antibody detection (100%) yet lowest specificity (40%). Similar results were obtained using the inhibition test as the reference standard. The sensitivity and specificity of the Kalon test versus the Western blot were 92% and 79%, respectively, and 80% and 82% versus the inhibition test. Using the inhibition test as the reference standard, the sensitivity of the Western blot appeared low (49%). In men in western Kenya who were HIV seronegative, the HerpeSelect and Kalon type-specific ELISAs had high sensitivities yet limited specificities using the Western blot as reference standard. Overall, the Kalon ELISA performed better than the HerpeSelect ELISA in these young men from Kisumu. Further understanding is needed for the interpretation of HSV-2 inhibition or ELISA test positive/ Western blot seronegative results. Before HSV-2 seropositivity may be reliably reported in selected areas of Africa, performance studies of HSV-2 serological assays in individual geographical areas are recommended.

  6. IgG and IgM western blot assay for diagnosis of congenital toxoplasmosis

    Directory of Open Access Journals (Sweden)

    Anderson S Machado

    2010-09-01

    Full Text Available The aim of this study was to evaluate the utility of western blot (WB analysis as a diagnostic tool for congenital toxoplasmosis in 215 newborn infants. The children were submitted to clinical examinations to assess macular, neurological and hearing signals. The WB results obtained were compared to the persistence of IgG antibodies at the end of 12 months, which is regarded as the "gold standard" diagnosis of congenital toxoplasmosis. Association between the WB results and the clinical signs presented by the infants was also assessed. Of the 215 children, 177 had a confirmed congenital toxoplasmosis diagnosis and 38 were uninfected. IgG-WB showed a sensitivity of 73.5% and a specificity of 97.4%. IgM-WB showed a sensitivity of 54.8% and a specificity of 94.7%. The IgG-WB and IgM-WB combination increased the sensitivity to 86.5%. The IgM-WB-positive children had a 1.4-fold greater risk of presenting active macular lesions than did those that were IgM-WB-negative. This study showed that the WB assay is a useful tool to confirm a diagnosis of congenital toxoplasmosis and that the IgM-WB-positive results can indicate active macular lesions in newborn infants.

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

    Directory of Open Access Journals (Sweden)

    Mohammad Zibaei

    2013-01-01

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

  8. Glyceraldehyde-3-phosphate dehydrogenase: a universal internal control for Western blots in prokaryotic and eukaryotic cells.

    Science.gov (United States)

    Wu, Yonghong; Wu, Min; He, Guowei; Zhang, Xiao; Li, Weiguang; Gao, Yan; Li, Zhihui; Wang, Zhaoyan; Zhang, Chenggang

    2012-04-01

    In the current study, we examined the expression level of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein in a number of organisms and the stability of GAPDH under various conditions. Our results revealed that GAPDH is present in multiple Escherichia coli strains, the yeast strain GS115, Caenorhabditis elegans, rat PC12 cells, and both mouse and rat brain. Furthermore, GAPDH was stably expressed under different concentrations of inducer and at different times of induction in E. coli (BL21) cells and yeast GS115 cells. Stable expression of GAPDH protein was also observed in C.elegans and PC12 cells that were treated with different concentrations of paraquat or sodium sulfite, respectively. In addition, we were able to detect and identify the endogenous gapA protein in E.coli via immunoprecipitation and MALDI-TOF-MS analysis. Endogenous gapA protein and exogenously expressed (subcloned) GAPDH proteins were detected in E. coli BL21 but not for gapC. With the exception of gapC in E. coli, the various isoforms of GAPDH possessed enzymatic activity. Finally, sequence analysis revealed that the GAPDH proteins were 76% identical, with the exception of E. coli gapC. Taken together, our results indicate that GAPDH could be universally used as an internal control for the Western blot analysis of prokaryotic and eukaryotic samples. Crown Copyright © 2012. Published by Elsevier Inc. All rights reserved.

  9. Serological diagnosis of North American Paragonimiasis by Western blot using Paragonimus kellicotti adult worm antigen.

    Science.gov (United States)

    Fischer, Peter U; Curtis, Kurt C; Folk, Scott M; Wilkins, Patricia P; Marcos, Luis A; Weil, Gary J

    2013-06-01

    Abstract. We studied the value of an IgG Western blot (WB) with Paragonimus kellicotti (Pk) antigen for diagnosis of North American paragonimiasis. The test was evaluated with sera from patients with Pk and Paragonimus westermani infections, with control sera from patients with other helminth infections, and sera from healthy Americans. All 11 proven Pk infection sera and two samples from suspected cases that were negative by P. westermani WB at the Centers for Disease Control and Prevention (CDC) contained antibodies to antigens at 34 kDa and at 21/23 kDa. Seven of 7 P. westermani sera contained antibodies to the 34 kDa antigen, but only 2 recognized the 21/23 kDa doublet. No control samples were reactive with these antigens. Antibody reactivity declined after praziquantel treatment. Thus, the P. kellicotti WB appears to be superior to P. westermani WB for diagnosing Pk infections, and it may be useful for assessing responses to treatment.

  10. An alternative strategy to western blot as a confirmatory diagnostic test for HIV infection.

    Science.gov (United States)

    Feng, Xia; Wang, Jibao; Gao, Zhiyun; Tian, Yu; Zhang, Ling; Chen, Huichao; Zhang, Tong; Xiao, Lin; Yao, Jun; Xing, Wenge; Qiu, Maofeng; Jiang, Yan

    2017-03-01

    In China, western blot (WB) is the recommended procedure for the diagnosis of HIV infection. However, this technique is time consuming and labor intensive, and its complexity restricts wide application in resource-limited regions. The aim of this study was to evaluate the efficacy of a dry blood spots (DBS)-urine paired enzyme-linked immunosorbent assay (ELISA) test, instead of WB, for HIV antibody detection. Plasma, DBS, and urine samples were collected from 1213 subjects from different populations. Two diagnostic testing strategies were conducted in parallel. The equivalence of the paired ELISA and WB strategies was assessed. A diagnosis of HIV was determined in 250 subjects according to the paired ELISA test, and in 249 according to the WB strategy. The discordant case was judged HIV-positive during follow-up. In total, 18 subjects were diagnosed with possible HIV using the paired ELISA test, among whom, 11 subjects tested negative with WB, and one was confirmed to be HIV-positive during follow-up. For the remaining 945 subjects, both strategies indicated a negative result. The kappa test indicated good conformity (kappa=0.954) between the two diagnostic strategies. The DBS-urine paired ELISA could be applied as an alternative to WB in HIV diagnosis, which would be valuable in resource-limited regions owing to the associated affordability and ease of use. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  11. Investigation of telomerase activity and apoptosis on invasive ductal carcinoma of the breast using immunohistochemical and Western blot methods.

    Science.gov (United States)

    Simsek, B C; Turk, B A; Ozen, F; Tuzcu, M; Kanter, M

    2015-08-01

    Invasive ductal carcinoma (IDC) comprises the largest group of breast cancers. This study aimed to investigate telomerase activity and apoptosis using immunohistochemical and Western blot methods. In total, 75 cases that had been diagnosed as IDC and 20 cases that had undergone a freezing procedure were included. The histological sections were stained with Bax, Bcl-2, hTERT and BNIP3. The ages of the patients, as well as their hormonal status and tumour sizes and grades were evaluated, as well as the staining characteristics of the antibodies in question. A decrease in Bcl-2 positivity and an increase in Bax positivity were found immunohistochemically with increasing tumour grades. The data obtained by western blot method showed that Bcl-2 was highest in grade 1 tumours although these results were not statistically significant. The relationship between estrogen and progesterone receptor positivity and Bcl-2 was statistically significant, suggesting there is hormonal control through apoptosis. BNIP3 was found to be decreased with increasing tumour grades. Similarly, BNIP3 was found to be having the lowest value in grade 3 tumours by western blot method. Furthermore, hTERT was found to be increased with increasing tumour grades. In the western blot method, hTERT increased nearly four-fold compared to the control. In addition, hTERT, which was seen in very high levels in tumours, may be a helpful cancer marker. Both hTERT and BNIP3 are important markers that can provide information about prognosis. Big improvements can be achieved in tumour progression control with new treatment modalities that stop telomerase activity and hypoxic cell death.

  12. A Western Blot Protocol for Detection of Proteins Heterologously Expressed in Xenopus laevis Oocytes.

    Science.gov (United States)

    Jørgensen, Morten Egevang; Nour-Eldin, Hussam Hassan; Halkier, Barbara Ann

    2016-01-01

    Oocytes of the African clawed frog, Xenopus laevis, are often used for expression and biochemical characterization of transporter proteins as the oocytes are particularly suitable for uptake assays and electrophysiological recordings. Assessment of the expression level of expressed transporters at the individual oocyte level is often desirable when comparing properties of wild type and mutant transporters. However, a large content of yolk platelets in the oocyte cytoplasm makes this a challenging task. Here we report a method for fast and easy, semiquantitative Western blot analysis of proteins heterologously expressed in Xenopus oocytes.

  13. 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.

  14. 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.

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

    Science.gov (United States)

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

    2012-03-06

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

  16. Standard loading controls are not reliable for Western blot quantification across brain development or in pathological conditions.

    Science.gov (United States)

    Goasdoue, Kate; Awabdy, Doreen; Bjorkman, Stella Tracey; Miller, Stephanie

    2016-02-01

    A frequently utilized method of data quantification in Western blot analysis is comparison of the protein of interest with a house keeping gene or control protein. Commonly used proteins include β-actin, glyceraldehyde 3 phosphate dehydrogenase (GAPDH), and α-tubulin. Various reliability issues have been raised when using this technique for data analysis-particularly when investigating protein expression changes during development and in disease states. In this study, we have demonstrated that β-actin, GAPDH, and α-tubulin are not appropriate controls in the study of development and hypoxic-ischemic induced damage in the piglet brain. We have also shown that using an in-house pooled standard, loaded on all blots is a reliable method for controlling interassay variability and data normalization in protein expression analysis. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  17. Western blot assay for quantitative and qualitative antigen detection in vaccine development.

    Science.gov (United States)

    Kumar, Sanjai; Zheng, Hong; Mahajan, Babita; Kozakai, Yukiko; Morin, Merribeth; Locke, Emily

    2014-05-01

    Immunological methods for quantitative measurement, antigenic characterization, and monitoring the stability of active immunogenic component(s) are a critical need in the vaccine development process. This unit describes an enhanced chemiluminescence-based western blot for quantitative detection of Plasmodium falciparum circumsporozoite protein (PfCSP), a major malaria candidate vaccine antigen. The most salient features of this assay are its high sensitivity and reproducibility; it can reliably detect ∼5 to 10 pg PfCSP expressed on native parasites or recombinantly expressed in Escherichia coli. Although described for a specific vaccine antigen, this assay should be applicable for any antigen-antibody combination for which relevant detection reagents are available. Detailed stepwise experimental procedures and methods for data acquisition and analysis are described. Copyright © 2014 John Wiley & Sons, Inc.

  18. 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

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

    Science.gov (United States)

    Eaton, Samantha L; Roche, Sarah L; Llavero Hurtado, Maica; Oldknow, Karla J; Farquharson, Colin; Gillingwater, Thomas H; Wishart, Thomas M

    2013-01-01

    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 that normalisation

  20. Discrepancies between a new highly sensitive Toxoplasma gondii ELISA assay and other reagents: interest of Toxo IgG Western blot.

    Science.gov (United States)

    Leslé, F; Touafek, F; Fekkar, A; Mazier, D; Paris, L

    2011-10-01

    Immunodiagnostic assays are commonly used to screen for maternal toxoplasmic seroconversion during pregnancy. The introduction to the market of a new highly sensitive IgG assay, the Elecsys Toxo IgG test, has resulted in discrepancy issues with other immunoassays because of a lack of standardisation. Western blot appears to be a good alternative gold standard to the dye test, as the latter is not routinely available. For the present prospective study, we compared the analytical performances of two immunoassays, Elecsys Toxo IgG (Roche Diagnostics) and Platelia Toxo IgG (Bio-Rad, Marnes la Coquette, France), to Toxo II IgG Western blot (LDBio, Lyon, France) using 231 consecutive sera with low or equivocal IgG titres. Of these 231 sera, 213 presented discrepancies, which showed the importance of a confirmation test. Of the Elecsys Toxo IgG-positive results, 100% were confirmed by the Western blot with a positive threshold of 30 IU/ml for Elecsys; in the equivocal area (1-30 IU/ml), Western blot is negative in 54% of cases. Our results suggest that the lower diagnostic cut-off of Platelia Toxo IgG should be further reduced. Our study indirectly confirms that monitoring, especially for pregnant women, must be done in the same laboratory using the same technique. The ability to diagnose very early seroconversion using Western blot merits further study.

  1. Evaluation of the Western blotting method for the diagnosis of congenital toxoplasmosis,

    Directory of Open Access Journals (Sweden)

    Jaqueline Dario Capobiango

    Full Text Available Abstract Objective: To evaluate the Western blotting method for the detection of IgG anti-Toxoplasma gondii (T. gondii (IgG-WB in the serum of children with suspected congenital toxoplasmosis. Methods: We accompanied 47 mothers with acquired toxoplasmosis in pregnancy and their children, between June of 2011 and June of 2014. The IgG-WB was done in house and the test was considered positive if the child had antibodies that recognized at least one band on IgG blots different from the mother's or with greater intensity than the corresponding maternal band, during the first three months of life. Results: 15 children (15.1% met the criteria for congenital toxoplasmosis and 32 (32.3% had the diagnosis excluded. The symptoms were observed in 12 (80.0% children and the most frequent were cerebral calcification in 9 (60.0%, chorioretinitis in 8 (53.3%, and hydrocephalus in 4 (26.6%. IgM antibodies anti-T. gondii detected by chemiluminescence (CL were found in 6 (40.0% children and the polymerase chain reaction (PCR for detection of T. gondii DNA was positive in 5 of 7 performed (71.4%. The sensitivity of IgG-WB was of 60.0% [95% confidence interval (CI 32.3-83.7%] and specificity 43.7% (95% CI 26.7-62.3%. The sensitivity of IgG-WB increased to 76.0 and 89.1% when associated to the research of IgM anti-T. gondii or PCR, respectively. Conclusions: The IgG-WB showed greater sensitivity than the detection of IgM anti-T. gondii; therefore, it can be used for the diagnosis of congenital toxoplasmosis in association with other congenital infection markers.

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

    Science.gov (United States)

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

    2012-03-01

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

  3. 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

  4. Variations in Western blot banding patterns of human T-cell lymphotropic virus type III/lymphadenopathy-associated virus.

    OpenAIRE

    Burke, D S; Redfield, R R; Putman, P; Alexander, S S

    1987-01-01

    Serum samples from 27 patients infected with human T-cell lymphotropic virus type III (14 with acquired immune deficiency syndrome [AIDS] and 13 with AIDS-related complex) were examined for antibodies to viral proteins by the Western blot method and with four different commercial solid-phase enzyme-linked immunosorbent assays (ELISAs). Virus-specific bands on blots at molecular masses of 64, 55, 53, 41, 31, 24, and 17 kilodaltons were observed. Rank correlation matrices were calculated to rel...

  5. Improved high-throughput quantification of luminescent microplate assays using a common Western-blot imaging system.

    Science.gov (United States)

    Hawkins, Liam J; Storey, Kenneth B

    2017-01-01

    Common Western-blot imaging systems have previously been adapted to measure signals from luminescent microplate assays. This can be a cost saving measure as Western-blot imaging systems are common laboratory equipment and could substitute a dedicated luminometer if one is not otherwise available. One previously unrecognized limitation is that the signals captured by the cameras in these systems are not equal for all wells. Signals are dependent on the angle of incidence to the camera, and thus the location of the well on the microplate. Here we show that: •The position of a well on a microplate significantly affects the signal captured by a common Western-blot imaging system from a luminescent assay.•The effect of well position can easily be corrected for.•This method can be applied to commercially available luminescent assays, allowing for high-throughput quantification of a wide range of biological processes and biochemical reactions.

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

    Science.gov (United States)

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

    2012-09-01

    Many CE-based technologies such as imaged capillary IEF, CE-SDS, CZE, and MEKC are well established for analyzing proteins, viruses, or other biomolecules such as polysaccharides. For example, imaged capillary isoelectric focusing (charge-based protein separation) and CE-SDS (size-based protein separation) are standard replacement methods in biopharmaceutical industries for tedious and labor intensive IEF and SDS-PAGE methods, respectively. Another important analytical tool for protein characterization is a Western blot, where after size-based separation in SDS-PAGE the proteins are transferred to a membrane and blotted with specific monoclonal or polyclonal antibodies. Western blotting analysis is applied in many areas such as biomarker research, therapeutic target identification, and vaccine development. Currently, the procedure is very manual, laborious, and time consuming. Here, we evaluate a new technology called Simple Western™ (or Simon™) for performing automated Western analysis. This new technology is based on CE-SDS where the separated proteins are attached to the wall of capillary by a proprietary photo activated chemical crosslink. Subsequent blotting is done automatically by incubating and washing the capillary with primary and secondary antibodies conjugated with horseradish peroxidase and detected with chemiluminescence. Typically, Western blots are not quantitative, hence we also evaluated the quantitative aspect of this new technology. We demonstrate that Simon™ can quantitate specific components in one of our vaccine candidates and it provides good reproducibility and intermediate precision with CV <10%. © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  7. The Prevalence and Significance of HTLV-I/II Seroindeterminate Western Blot Patterns

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    Yoshimi Akahata

    2011-08-01

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

  8. A new Western blot assay for the detection of porcine cytomegalovirus (PCMV).

    Science.gov (United States)

    Plotzki, Elena; Keller, Martina; Ivanusic, Daniel; Denner, Joachim

    2016-10-01

    Porcine cytomegalovirus (PCMV) may be harmful for human recipients if xenotransplantation using pig cell, tissue or organ will be performed transmitting the virus from donor pigs to human recipients. PCMV is widespread in pigs and closely related to human pathogenic herpesviruses, however there are no data concerning infection of humans. In contrast, recently it had been shown that transplantation of organs from pigs infected with PCMV into non-human primate recipients resulted in a significant reduction of the survival time compared with the transplantation of organs from uninfected pigs. To prevent transmission of PCMV in future pig to human xenotransplantations, sensitive and specific detection methods should be used. Here a new Western blot assay using recombinant proteins corresponding to two domains of the glycoprotein gB of PCMV is described. With this assay, the presence of PCMV-specific antibodies in different pig breeds was analysed. Antibodies were detected in a high percentage of animals, in one breed up to 85%. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Utility of Bartonella henselae IgM Western Blot Bands for Serodiagnosis of Cat Scratch Disease.

    Science.gov (United States)

    Otsuyama, Ken-Ichiro; Tsuneoka, Hidehiro; Yoshidomi, Hiroka; Haraguchi, Mio; Yanagihara, Masashi; Tokuda, Nobuko; Nojima, Junzo; Ichihara, Kiyoshi

    2018-01-01

    We evaluated the utility of Western blot (WB) bands of Bartonella henselae in detecting anti-B. henselae immunoglobulin M (IgM) for serodiagnosis of cat scratch disease (CSD). IgM band patterns were examined using sera from 92 patients clinically suspected of having CSD and from 130 healthy individuals. Positive WB bands were observed in 49 (53.5%) of the 92 patient sera. Three bands at 8 to 10, 31 to 35, and 70 kDa were regarded as relevant for B. henselae because all of the positive sera yielded at least one of the three bands, and none of the healthy control sera showed reactivity to any of them. In contrast, the positive rate of the patient sera by conventional indirect fluorescence antibody assay (IFA) for B. henselae IgM was 28.3% (26/92) among the patients. These finding suggest that the IgM-WB assay, although cumbersome to perform, can be used for confirmatory diagnosis of CSD with no false positivity in the control sera. Purification of proteins in the specific bands may contribute to the development of an IgM enzyme-linked immunosorbent assay (IgM-ELISA) with improved specificity and sensitivity. Copyright © 2017 American Society for Microbiology.

  10. Novel chemiluminescent Western blot blocking and antibody incubation solution for enhanced antibody-antigen interaction and increased specificity.

    Science.gov (United States)

    Schwartz, Kimberly; Bochkariov, Dmitry

    2017-10-01

    Western blotting is a ubiquitous tool used in protein and molecular biology research, providing information about the presence, size, relative abundance, and state of a protein in a mixture. First, the proteins in a sample are separated by size using SDS-PAGE then transferred onto a membrane for detection with a set of primary and secondary antibodies. High-quality Western data requires high signal-to-noise ratios, which depend upon reduction of nonspecific antibody interactions. Blocking is a critical step in the Western blot method as it prevents the antibodies from binding nonspecifically to the membrane and irrelevant proteins. A solution of nonfat dry milk (NFDM) in physiological buffer is commonly used for this purpose, but does not perform well with every type of antibody and is not optimal for low-abundance proteins. We present a novel blocking solution for chemiluminescent Western blots, AdvanBlock™-chemi, which outperforms NFDM in experiments with 20 unique antibodies by increasing signal-to-noise ratios and minimizing nonspecific binding. This solution enhances protein detection by Western blot and provides consistent results for detection of low abundant and modified proteins. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  11. Western blot analysis of sera from dogs with suspected food allergy.

    Science.gov (United States)

    Favrot, Claude; Linek, Monika; Fontaine, Jacques; Beco, Luc; Rostaher, Ana; Fischer, Nina; Couturier, Nicolas; Jacquenet, Sandrine; Bihain, Bernard E

    2017-04-01

    Food allergy is often suspected in dogs with clinical signs of atopic dermatitis. This diagnosis is confirmed with an elimination diet and a subsequent challenge with regular food. Laboratory tests for the diagnosis of food allergy in dogs are unreliable and/or technically difficult. Cyno-DIAL ® is a Western blot method that might assist with the selection of an appropriate elimination diet. To evaluate the performance of Cyno-DIAL ® for the selection of an elimination diet and diagnosis of food allergy. Thirty eight dogs with atopic dermatitis completed an elimination diet. Combining the results of the diet trials and the challenges, 14 dogs were classified as food allergic (FA), 22 as nonfood-allergic and two as ambiguous cases. Amongst all dogs and amongst dogs with a clinical diagnosis of FA, 3% and 7% (respectively) were positive to Royal Canin Anallergenic ® , Vet-Concept Kanguru ® or Vet-Concept Dog Sana ® ; 8% and 7% to Hill's d/d Duck and Rice ® ; 8% and 21% to Hill's z/d Ultra Allergen Free ® ; 53% and 64% to Eukanuba Dermatosis FP ® ; and 32% and 43% to a home-cooked diet of horse meat, potatoes and zucchini. The specificity and sensitivity of Cyno-DIAL ® for diagnosing food allergy were 73% and 71%, respectively. Although Cyno-DIAL ® was considered potentially useful for identifying appropriate foods for elimination diet trials, it cannot be recommended for the diagnosis of food allergy. The Cyno-DIAL ® test performed better than some previously evaluated ELISA-based tests. © 2017 ESVD and ACVD.

  12. Enrichment of PrPSc in Formalin Fixed Paraffin Embedded Tissues Prior to Analysis by Western Blot

    Science.gov (United States)

    Diagnosis of prion disease is primarily through immunodetection of the infectious agent. Typically, 2 distinct procedures are recommended for a definitive diagnosis with immunohistochemistry and Western blot providing the most information as to the specific isolate in question. In the past these app...

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

    Science.gov (United States)

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

    2015-12-01

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

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

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    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.

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

    Science.gov (United States)

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

    2013-09-01

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

  16. Comparison of reflectance and transmission densitometry, using document and laser scanners, for quantitation of stained Western blots.

    Science.gov (United States)

    Tarlton, J F; Knight, P J

    1996-05-15

    The quantitation of stained Western blots by reflectance and transmission scanning was explored using a blot of monoclonal immunoglobulin G probed with anti-mouse antibody. The so-called "scanned absorbance" output from a document scanner was found to be directly proportional to the fraction of light absorbed rather than obeying the logarithmic relationship expected for true spectrophotometric absorbance. This explains observations in the literature of a strongly curved relation between loading and "absorbance." Laser transmission densitometry of a blot immersed in a clarifying solvent mixture showed that peak area was linearly related to loading over a wide range. In reflectance mode the document scanner also gave equally linear quantitation of dry blots, providing that a logarithmic correction curve was applied during scanning. It was found advantageous to interpose a red acetate filter sheet between the blot and the scanning table to aid detection of weakly stained bands. The document scanner gave less satisfactory results when used in transmission mode on a clarified blot because weak bands were poorly quantitated.

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

    Science.gov (United States)

    Rivera-Torres, José

    2015-01-01

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

  18. Diagnostic efficacy of Brucella abortus strain RB51 in experimentally inoculated Sprague-Dawley rats using western blot assay.

    Science.gov (United States)

    Rahman, Siddiqur; Baek, Byeong Kirl

    2008-10-01

    To investigate the diagnosis and efficacy of Brucella abortus strain RB51 (SRB51) in experimentally inoculated Sprague-Dawley (SD) rat using western blot assay. Female SD rats were orally administered with 1.0 x 10(7) colony forming unit (cfu) suspension of SRB51 and half of these SD rats were challenged at 4 weeks post inoculation with 1.0 x 10(9) cfu suspension of B. abortus biotype 1 isolated in South Korea. Sera of SD rats were monitored at regular intervals by western blot assay using whole cell antigen of B. abortus strain 1119-3 (S1119-3). The bacteriological examination of blood and clinical examination of the rats were also performed. There were several bands at 120, 70, 45, 30, 20 kDa and clear specific bands were found after vaccination (20, 70 kDa) and challenge (15, 20, 45, 70, 120 kDa). The highest immune response was observed in sera 4 weeks post SRB51 vaccination. SRB51 was recovered from the blood of all of SRB51 inoculated rats until one week post vaccination and there were no clinical signs in that inoculated rats. It is concluded that the SRB51 elicits antigen specific immunity in SD rats based on western blot assay.

  19. Serodiagnosis of grass carp reovirus infection in grass carp Ctenopharyngodon idella by a novel Western blot technique.

    Science.gov (United States)

    He, Yongxing; Jiang, Yousheng; Lu, Liqun

    2013-12-01

    Frequent outbreaks of grass carp hemorrhagic disease, caused by grass carp reovirus (GCRV) infection, pose as serious threats to the production of grass carp Ctenopharyngodon idella. Although various nucleic acids-based diagnostic methods have been shown effective, lack of commercial monoclonal antibody against grass carp IgM has impeded the development of any reliable immunoassays in detection of GCRV infection. The present study describes the preparation and screening of monoclonal antibodies against the constant region of grass carp IgM protein, and the development of a Western blot (WB) protocol for the specific detection of antibodies against outer capsid VP7 protein of GCRV that serves as antibody-capture antigen in the immunoassay. In comparison to a conventional RT-PCR method, validity of the WB is further demonstrated by testing on clinical fish serum samples collected from a grass carp farm in Jiangxi Province during disease pandemic in 2011. In conclusion, the WB technique established in this study could be employed for specific serodiagnosis of GCRV infection. Copyright © 2013 Elsevier B.V. All rights reserved.

  20. Sensitive Western-Blot Analysis of Azide-Tagged Protein Post Translational Modifications Using Thermoresponsive Polymer Self-Assembly.

    Science.gov (United States)

    Liu, Tong; Zhang, Wanjun; Zhang, Zheng; Chen, Mingli; Wang, Jianhua; Qian, Xiaohong; Qin, Weijie

    2018-02-06

    Western-blot (WB) is a powerful analytical technique for protein identification in complex biological samples and has been widely used in biological studies for decades. Detection specificity and sensitivity of WB largely relies on quality of the antibodies and performance of the conjugated HRP. However, the application of WB analysis for the detection of protein post-translational modifications (PTMs) is hampered by the low abundance of protein PTMs and by the limited availability of antibodies that specifically differentiate various kinds of PTMs from their protein substrates. Therefore, new recognition mechanisms and signal amplification strategies for WB analysis of protein PTMs is in high demand. In this work, we prepared a soluble polymer that detects various azide-tagged PTM proteins in WB analysis using triarylphosphine and HRP modified thermoresponsive polymer. Specific and efficient detection of azide-tagged PTM protein is achieved via the bioorthogonal reaction between azide and triarylphosphine. More importantly, the chemiluminiscent signal in the WB analysis is largely amplified by the temperature induced self-assembly of numerous thermoresponsive polymer chains carrying multiple HRPs. As a result, approximately 100 times more sensitive detection than commercial antibodies is achieved by this method using standard PTM proteins. Though, this new reagent does not directly detect native PTMs in cell, tissue or blood samples, it still has important application potential in protein PTM studies, considering the wide availability of azide-tagging techniques to a variety of PTMs.

  1. 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.

  2. Avoiding pitfalls of internal controls: validation of reference genes for analysis by qRT-PCR and Western blot throughout rat retinal development.

    Science.gov (United States)

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

    2012-01-01

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

  3. Western blotting using Strongyloides ratti antigen for the detection of IgG antibodies as confirmatory test in human strongyloidiasis

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    Luciana Pereira Silva

    2003-07-01

    Full Text Available The present study was conducted to evaluate the frequency of antigenic components recognized by serum IgG antibodies in Western blotting (WB using a Strongyloides ratti larval extract for the diagnosis of human strongyloidiasis. In addition, the WB results were compared to the enzyme-linked immunosorbent assay (ELISA and the indirect immunofluorescence antibody test (IFAT results. Serum samples of 180 individuals were analyzed (80 with strongyloidiasis, 60 with other intestinal parasitoses, and 40 healthy individuals. S. ratti was obtained from fecal culture of experimentally infected Rattus rattus. For IFAT, S. ratti larvae were used as antigen and S. ratti larval antigenic extracts were employed in WB and ELISA. Eleven S. ratti antigenic components were predominantly recognized by IgG antibodies in sera of patients with strongyloidiasis. There was a positive concordance for the three tests in 87.5% of the cases of strongyloidiasis. The negative concordance in the three tests was 94% and 97.5%, in patients with other intestinal parasitoses and healthy individuals, respectively. In cases of positive ELISA and negative IFAT results, diagnosis could be confirmed by WB. ELISA, IFAT, and WB using S. ratti antigens showed a high rate of sensitivity and specificity. In conclusion, WB using S. ratti larval extract was able to recognize 11 immunodominant antigenic components, showing to be a useful tool to define the diagnosis in cases of equivocal serology.

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

    Directory of Open Access Journals (Sweden)

    Haitham A. Badr

    2015-12-01

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

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

    Science.gov (United States)

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

    2010-11-01

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

  6. 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 (IgG1, 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.

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

    Science.gov (United States)

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

    2017-01-01

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

  8. 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

  9. 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.

  10. Development of enzyme immunoassays (ELISA and Western blot) for the serological diagnosis of dermatophytosis in symptomatic and asymptomatic cats.

    Science.gov (United States)

    Santana, Aline Elisa; Taborda, Carlos Pelleschi; Severo, Julia So; Rittner, Glauce Mary Gomes; Muñoz, Julian Esteban; Larsson, Carlos Eduardo; Larsson, Carlos Eduardo

    2018-01-01

    Dermatophytosis is the most common fungal infection in cats worldwide and plays an important role in both animal and human health due to their high zoonotic potential. Effective screening is a strong preventive measure and the fungal culture is quite useful but requires full laboratorial experience and it takes a long time to obtain the result. A rapid and accurate screening test for dermatophytosis in cats is crucial for the effective control of disease outbreaks. The aim of this study was to develop and evaluate the diagnostic efficacy of enzyme immunoassays (ELISA and Western blot [WB]) for the rapid and precise diagnosis of dermatophytosis in cats. Seventy cats of various ages were divided into three groups: S (symptomatic, n = 20), AS (asymptomatic, n = 30), and N (negative, n = 20). All animals were submitted to fungal culture and blood samples for carrying out the serological tests. A significant difference (P < 0.05) was found between IgG-specific levels of sera of Microsporum canis positive and negative animals. There was no statistic difference between groups symptomatic and asymptomatic. The ELISA test showed sensitivity of 94% and specificity of 75%. Receiver operating characteristic (ROC) analysis also showed higher diagnostic accuracy (AUC 0.925). The WB technique detected 13 bands, and the 50 kDa protein was considered the most immunogenic protein, observing reactivity in 83.3% in the symptomatic group and 66.6% in the asymptomatic group. The study concluded that ELISA and WB were useful tools to reliably detect cats that have been exposed to M. canis. © The Author 2017. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

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

    Science.gov (United States)

    Seyyedtabaei, Seyyed Javad; Rostami, Ali; Haghighi, Ali; Mohebali, Mehdi; Kazemi, Bahram; Fallahi, Shirzad; Spotin, Adel

    2017-01-01

    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. 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. 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. WB could be useful for screening and serodiagnosis of CL and VL in epidemiologic studies in endemic areas.

  12. [Western blot technique standardization for specific diagnosis of Chagas disease using excretory-secretory antigens of Trypanosoma cruzi epimastigotes].

    Science.gov (United States)

    Escalante, Hermes; Jara, César; Davelois, Kelly; Iglesias, Miguel; Benites, Adderly; Espinoza, Renzo

    2014-01-01

    Evaluate the effectiveness of Western Blot for the specific diagnosis of Chagas disease using excretory-secretory antigens of Trypanosoma cruzi epimastigotes. Antigens were obtained after twenty hours of incubation in Eagle’s Minimum Essential Medium, which were prepared at a protein concentration of 0.2 ug/uL to be faced with 10 mL pool of serum from patients with Chagas disease and a conjugated anti-IgG labeled with peroxidase. The presence of the following antigens was observed: 10, 12, 14, 15, 19, 20, 23, 26, 30, 33, 36, 40, 42, 46, 58, 63, 69, 91, 100, and 112 kDa; of which antigens of 10, 12, 14, 15, 19, 20, 23, and 26 kDa were considered to be specific using pools of serum from patients with other parasitosis and serum from people with no parasites. The sensitivity of the technique was assessed using individual serum from 65 patients with Chagas disease; and the specificity with serum from 40 patients with other parasitosis, and serums from five people who did not have parasites. The technique has a sensitivity of 95.4% in the detection of one to eight specific bands, a specificity of 100%, a positive predictive value of 100%, and a negative predictive value of 93.7%. Western Blot technique with excretory-secretory antigens of T. cruzi epimastigotes is effective in the diagnosis of Chagas disease in Peru; therefore, it can be used as a confirmatory test.

  13. Comparison of a serum indirect fluorescent antibody test with two Western blot tests for the diagnosis of equine protozoal myeloencephalitis.

    Science.gov (United States)

    Duarte, Paulo C; Daft, Barbara M; Conrad, Patricia A; Packham, Andrea E; Gardner, Ian A

    2003-01-01

    A serum indirect fluorescent antibody test (IFAT) was compared with a Western blot (WB) and a modified Western blot (mWB) for diagnosis of equine protozoal myeloencephalitis (EPM). Using receiver-operating characteristic (ROC) analysis, the area under the curve of the IFAT was greater than the areaunder the curves of the WB and the mWB (P = 0.025 and P = 0.044, respectively). There was no statistically significant difference between the areas under the curves of the WBs (P > 0.05). On the basis of an arbitrarily chosen cut-off titer for a positive test result of 1:80 for the IFAT and interpreting weak positive WB results as positive test results, the sensitivities and 95% confidence intervals (CI) of all 3 tests were identical and equal to 88.9% (51.8-99.7%). The specificities and 95% CIs of the IFAT, WB, and mWB test were 100% (91-100%), 87.2% (72.6-95.7%), and 69.2% (52.4-83%), respectively. The overall accuracy of the IFAT was shown to be better than that of the WBs and, therefore, the test has potential for use in the diagnosis of EPM caused by Sarcocystis neurona.

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

    Science.gov (United States)

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

    2017-02-01

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

  15. Evaluation of ELISA coupled with Western blot as a surveillance tool for Trichinella infection in wild boar (Sus scrofa).

    Science.gov (United States)

    Cuttell, Leigh; Gómez-Morales, Maria Angeles; Cookson, Beth; Adams, Peter J; Reid, Simon A; Vanderlinde, Paul B; Jackson, Louise A; Gray, C; Traub, Rebecca J

    2014-01-31

    Trichinella surveillance in wildlife relies on muscle digestion of large samples which are logistically difficult to store and transport in remote and tropical regions as well as labour-intensive to process. Serological methods such as enzyme-linked immunosorbent assays (ELISAs) offer rapid, cost-effective alternatives for surveillance but should be paired with additional tests because of the high false-positive rates encountered in wildlife. We investigated the utility of ELISAs coupled with Western blot (WB) in providing evidence of Trichinella exposure or infection in wild boar. Serum samples were collected from 673 wild boar from a high- and low-risk region for Trichinella introduction within mainland Australia, which is considered Trichinella-free. Sera were examined using both an 'in-house' and a commercially available indirect-ELISA that used excretory-secretory (E/S) antigens. Cut-off values for positive results were determined using sera from the low-risk population. All wild boar from the high-risk region (352) and 139/321 (43.3%) of the wild boar from the low-risk region were tested by artificial digestion. Testing by Western blot using E/S antigens, and a Trichinella-specific real-time PCR was also carried out on all ELISA-positive samples. The two ELISAs correctly classified all positive controls as well as one naturally infected wild boar from Gabba Island in the Torres Strait. In both the high- and low-risk populations, the ELISA results showed substantial agreement (k-value=0.66) that increased to very good (k-value=0.82) when WB-positive only samples were compared. The results of testing sera collected from the Australian mainland showed the Trichinella seroprevalence was 3.5% (95% C.I. 0.0-8.0) and 2.3% (95% C.I. 0.0-5.6) using the in-house and commercial ELISA coupled with WB respectively. These estimates were significantly higher (P<0.05) than the artificial digestion estimate of 0.0% (95% C.I. 0.0-1.1). Real-time PCR testing of muscle from

  16. Prevalence of indeterminate human immunodeficiency virus western blot results in pregnant women attended at a public hospital in Presidente Prudente, Brazil

    National Research Council Canada - National Science Library

    Cremonezi, Denise; Mesquita, Paulo Eduardo de; Romão, Marisa Menezes; Prestes-Carneiro, Luiz Euribel

    2005-01-01

    .... In Brazil, the prevalence of HIV infection among pregnant women is less than 1%. Therefore, the positive predictive value of an HIV EIA test tends to be lower than the more frequent indeterminate Western blot result...

  17. 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

    Reference proteins (RP) or the total protein (TP) loaded is used to correct for uneven loading and/or transfer in Western blotting. However, the signal sensitivity and the influence of physiological conditions may question the normalization methods. Therefore, three widely used reference proteins...... and differences in muscle fiber type. The novel SF technology adds lower variation to the results compared with the existing methods for correcting for loading inaccuracy in Western blotting of human skeletal muscle in applied physiology....

  18. Prevalence of indeterminate human immunodeficiency virus western blot results in pregnant women attended at a public hospital in Presidente Prudente, Brazil.

    Science.gov (United States)

    Cremonezi, Denise; Mesquita, Paulo Eduardo de; Romão, Marisa Menezes; Prestes-Carneiro, Luiz Euribel

    2005-12-01

    The AIDS epidemic is spreading rapidly among women worldwide, offering increasing opportunities for vertical transmission of HIV. In Brazil, the prevalence of HIV infection among pregnant women is less than 1%. Therefore, the positive predictive value of an HIV EIA test tends to be lower than the more frequent indeterminate Western blot result. Pregnant women receiving antenatal care, from 2000 to 2004, at a public secondary hospital in the city of Presidente Prudente, São Paulo, Brazil, were systematically screened for HIV by means of two distinct EIA tests, in order to determine the prevalence of indeterminate Western blot results among pregnant women showing discordance in both HIV EIA tests and indirect immunofluorescence assay. Confirmatory indirect immunofluorescence was performed on material for all women with positive results in both EIA tests. Whenever there were positive results in EIA and IIA, the applicant was retested by the initial screening assay. Only those not showing concordance in results in EIA and IAA had a Western blot performed. The viral load was measured in pregnant women with positive or indeterminate Western blot results. Out of 9,786 sera, 105 (1.0%) were positive in the two HIV EIA screening tests, confirmed by indirect immunofluorescence. Among these women, Western blot was interpreted as indeterminate in 11 (0.1%) cases and their viral load was HIV indeterminate Western blots in pregnant women from Presidente Prudente and the surrounding region; none of these pregnant women had positive HIV viral loads.

  19. 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.’

  20. Quantitative analysis of a biopharmaceutical protein in cell culture samples using automated capillary electrophoresis (CE) western blot.

    Science.gov (United States)

    Xu, Dong; Marchionni, Kentaro; Hu, Yunli; Zhang, Wei; Sosic, Zoran

    2017-10-25

    An effective control strategy is critical to ensure the safety, purity and potency of biopharmaceuticals. Appropriate analytical tools are needed to realize such goals by providing information on product quality at an early stage to help understanding and control of the manufacturing process. In this work, a fully automated, multi-capillary instrument is utilized for size-based separation and western blot analysis to provide an early readout on product quality in order to enable a more consistent manufacturing process. This approach aims at measuring two important qualities of a biopharmaceutical protein, titer and isoform distribution, in cell culture harvest samples. The acquired data for isoform distribution can then be used to predict the corresponding values of the final drug substance, and potentially provide information for remedy through timely adjustment of the downstream purification process, should the expected values fall out of the accepted range. Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.

  1. Development, validation, and pilot application of a semiquantitative Western blot analysis and an ELISA for bovine adiponectin.

    Science.gov (United States)

    Mielenz, M; Mielenz, B; Singh, S P; Kopp, C; Heinz, J; Häussler, S; Sauerwein, H

    2013-04-01

    Adiponectin is an adipose tissue-derived glycoprotein circulating as highly abundant multimers. It regulates glucose metabolism and insulin sensitivity. In ruminants, valid data about serum concentrations and tissue-specific protein expression are lacking, and we, therefore, aimed to generate a polyclonal antibody against bovine adiponectin to apply it in immunodetection. The specificity of the purified anti-adiponectin antibody was established by Western blot analysis with the use of reducing and denaturing conditions applied to both the purified protein and the bovine serum samples. Besides bovine serum, the applicability of the antibody for immunodetection of adiponectin was confirmed for the supernatant fluid of in vitro-differentiated bovine adipocytes, for protein extracts from bovine adipose tissue, and also in a multispecies comparison: bands comparable in size with monomeric bovine adiponectin were obtained under denaturing conditions in serum of camel, horse, human, mouse, pig, roe deer, and sheep. In addition, when used in immunohistochemistry on bovine adipose tissue sections, a characteristic adipocyte-specific staining pattern was obtained with this antibody. The antibody was used for establishing a semiquantitative Western blot procedure and the development of an ELISA. Both methods were extensively validated and were first applied to characterize the serum adiponectin concentrations in multiparous dairy cows during the transition from pregnancy to lactation, that is, 3 wk before until 5 wk after calving. With both assays a time effect (P = 0.017, P = 0.026, respectively) with lowest values at the day of parturition was observed. We thus established 2 useful tools to validly assess bovine adiponectin at the protein level. Copyright © 2013 Elsevier Inc. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    Greenlee Justin J

    2011-09-01

    Full Text Available Abstract Background Three distinct forms of bovine spongiform encephalopathy (BSE, defined as classical (C-, low (L- or high (H- type, have been detected through ongoing active and passive surveillance systems for the disease. The aim of the present study was to compare the ability of two sets of immunohistochemical (IHC and Western blot (WB BSE confirmatory protocols to detect C- and atypical (L- and H-type BSE forms. Obex samples from cases of United States and Italian C-type BSE, a U.S. H-type and an Italian L-type BSE case were tested in parallel using the two IHC sets and WB methods. Results The two IHC techniques proved equivalent in identifying and differentiating between C-type, L-type and H-type BSE. The IHC protocols appeared consistent in the identification of PrPSc distribution and deposition patterns in relation to the BSE type examined. Both IHC methods evidenced three distinct PrPSc phenotypes for each type of BSE: prevailing granular and linear tracts pattern in the C-type; intraglial and intraneuronal deposits in the H-type; plaques in the L-type. Also, the two techniques gave comparable results for PrPSc staining intensity on the C- and L-type BSE samples, whereas a higher amount of intraglial and intraneuronal PrPSc deposition on the H-type BSE case was revealed by the method based on a stronger demasking step. Both WB methods were consistent in identifying classical and atypical BSE forms and in differentiating the specific PrPSc molecular weight and glycoform ratios of each form. Conclusions The study showed that the IHC and WB BSE confirmatory methods were equally able to recognize C-, L- and H-type BSE forms and to discriminate between their different immunohistochemical and molecular phenotypes. Of note is that for the first time one of the two sets of BSE confirmatory protocols proved effective in identifying the L-type BSE form. This finding helps to validate the suitability of the BSE confirmatory tests for BSE

  3. Evaluation of the Bio-Rad Multispot HIV-1/HIV-2 Rapid Test as an alternative to Western blot for confirmation of HIV infection.

    Science.gov (United States)

    Cárdenas, Ana María; Baughan, Eleonore; Hodinka, Richard L

    2013-12-01

    In the United States, a new HIV diagnostic algorithm has been proposed that uses an HIV-1/HIV-2 antibody differentiation immunoassay instead of Western blot or immunofluoresence for confirmatory testing. To evaluate the Multispot HIV-1/HIV-2 Rapid Test (Multispot) as an alternative to Western blot analysis for confirmation of HIV infection. A series of 205 serum and plasma specimens positive for HIV-1 or HIV-2 were used to compare the performance of Multispot to a standard HIV-1 Western blot. Positive samples included 63 specimens from patients>18 months of age, 33 proficiency survey specimens, and 109 specimens from nine commercial seroconversion and performance panels. In addition, 63 specimens from 51 HIV-exposed, uninfected children≤18 months of age in various stages of seroreversion and 192 HIV-negative samples were tested. Specimens were initially screened using a 4th generation HIV Ag/Ab Combo assay. Multispot readily discriminated between individuals with HIV-1 or HIV-2 infection and those who were uninfected. Of the 205 samples repeatedly reactive by the 4th generation screening assay, infection status was correctly confirmed by Multispot in 83.9% (172/205) compared to 68.8% (141/205) for Western blot. Multispot detected HIV-1 earlier in 27.6% of low-titer antibody specimens called indeterminate by Western blot, and effectively reduced the number of indeterminate results in seroreverting HIV-1 exposed, uninfected infants and for HIV-2 infections misinterpreted as indeterminate or positive by HIV-1 Western blot. Multispot offers speed and simplicity over Western blot and has an excellent performance for differentiation and confirmation of antibodies to HIV-1 and HIV-2. Copyright © 2013 Elsevier B.V. All rights reserved.

  4. 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.

  5. Confirmation of the immunoreactivity of monoclonal anti-human C-terminal EGFR antibodies in bronze Corydoras Corydoras aeneus (Callichthyidae Teleostei) by Western Blot method.

    Science.gov (United States)

    Mytych, Jennifer; Satora, Leszek; Kozioł, Katarzyna

    2017-12-12

    Bronze corydoras (Corydoras aeneus) uses the distal part of the intestine as accessory respiratory organ. Our previous study showed the presence of epidermal growth factor receptor (EGFR) cytoplasmic domain in the digestive tract of the bronze corydoras. In this study, using Western Blot method, we validated the results presented in the previous research. In detail, results of Western Blot analysis on digestive and respiratory part of bronze corydoras intestine homogenates confirmed the immunoreactivity of anti-cytoplasmic domain (C-terminal) human EGFR antibodies with protein band of approximately 180kDa (EGFR molecular weight). This indicates a high homology of EGFR domain between these species and the possibility of such antibody use in bronze corydoras. Statistically significantly higher EGFR expression was observed in the respiratory part of intestine when compared to the digestive part. This implies higher proliferation activity and angiogenesis of epithelium in this part of intestine, creating conditions for air respiration. Therefore, Corydoras aeneus may be considered as a model organism for the molecular studies of the mechanisms of epithelial proliferation initiation and inhibition depending on hypoxia and normoxia. Copyright © 2017. Published by Elsevier GmbH.

  6. Characterization of 65 epitope-specific dystrophin monoclonal antibodies in canine and murine models of duchenne muscular dystrophy by immunostaining and western blot.

    Science.gov (United States)

    Kodippili, Kasun; Vince, Lauren; Shin, Jin-Hong; Yue, Yongping; Morris, Glenn E; McIntosh, Mark A; Duan, Dongsheng

    2014-01-01

    Epitope-specific monoclonal antibodies can provide unique insights for studying cellular proteins. Dystrophin is one of the largest cytoskeleton proteins encoded by 79 exons. The absence of dystrophin results in Duchenne muscular dystrophy (DMD). Over the last two decades, dozens of exon-specific human dystrophin monoclonal antibodies have been developed and successfully used for DMD diagnosis. Unfortunately, the majority of these antibodies have not been thoroughly characterized in dystrophin-deficient dogs, an outstanding large animal model for translational research. To fill the gap, we performed a comprehensive study on 65 dystrophin monoclonal antibodies in normal and dystrophic dogs (heart and skeletal muscle) by immunofluorescence staining and western blot. For comparison, we also included striated muscles from normal BL10 and dystrophin-null mdx mice. Our analysis revealed distinctive species, tissue and assay-dependent recognition patterns of different antibodies. Importantly, we identified 15 antibodies that can consistently detect full-length canine dystrophin in both immunostaining and western blot. Our results will serve as an important reference for studying DMD in the canine model.

  7. A research design for the quantification of the neuropeptides substance p and calcitonin gene-related Peptide in rat skin using Western blot analysis.

    Science.gov (United States)

    Lapin, Guilherme Abbud Franco; Hochman, Bernardo; Nishioka, Michele Akemi; Maximino, Jessica Ruivo; Chadi, Gerson; Ferreira, Lydia Masako

    2015-06-01

    To describe and standardize a protocol that overcomes the technical limitations of Western blot (WB) analysis in the quantification of the neuropeptides substance P (SP) and calcitonin gene-related peptide (CGRP) following nociceptive stimuli in rat skin. Male Wistar rats (Rattus norvegicus albinus) weighing 250 to 350 g were used in this study. Elements of WB analysis were adapted by using specific manipulation of samples, repeated cycles of freezing and thawing, more thorough maceration, and a more potent homogenizer; increasing lytic reagents; promoting greater inhibition of protease activity; and using polyvinylidene fluoride membranes as transfer means for skin-specific protein. Other changes were also made to adapt the WB analysis to a rat model. University research center. Western blot analysis adapted to a rat model. This research design has proven effective in collecting and preparing skin samples to quantify SP and CGRP using WB analysis in rat skin. This study described a research design that uses WB analysis as a reproducible, technically accessible, and cost-effective method for the quantification of SP and CGRP in rat skin that overcomes technical biases.

  8. Determining the cleavage site for the mature antimicrobial peptide of Nile tilapia β-defensin using 2D electrophoresis, western blot, and mass spectrometry analysis.

    Science.gov (United States)

    Chang, Chin-I; Chen, Li-Hao; Hu, Yeh-Fang; Wu, Chia-Che; Tsai, Jyh-Ming

    2017-03-01

    Several proteomic techniques were used to determine the cleavage site of the mature antimicrobial peptide of Nile tilapia β-defensin. The computer-predicted Nile tilapia β-defensin ( 25 ASFPWSCLSLSGVCRKVCLPTELFFGPLGCGKGSLCCVSHFL 66 ) composed of 42 amino acids was chemically synthesized and prepared to produce an antibody for Western blotting. Total proteins from the skin of the Nile tilapia were separated on two-dimensional electrophoresis, and the spot of Nile tilapia β-defensin was recognized using Western blot analysis. It was then excised and extracted from the gel. The precise molecular mass of this spot was determined by LC-MS/MS spectrometry. Four major peptides were discovered, with molecular weights of 4293.2 Da, 4306.5 Da, 4678.9 Da, and 4715.0 Da. The calculated mass of the 40-amino-acid sequence ( 27 FPWSCLSLSGVCRKVCLPTELFFGPLGCGKGSLCCVSHFL 66 ) of Nile tilapia β-defensin starting from Phe27 and ending with Leu66 was 4293.18 Da, which completely matched the 4293.2 Da peptide that was obtained from the mass spectrometry analysis. This result confirmed that the cleavage site for the mature C-terminal Nile tilapia β-defensin is at residue Ser26-Phe27, not at Ala24-25 as predicted by computer analysis. This study provides a simple but reliable model to determine the cleavage site for a mature antimicrobial peptide. Copyright © 2017 Elsevier Ltd. All rights reserved.

  9. Prevalence of indeterminate human immunodeficiency virus western blot results in pregnant women attended at a public hospital in Presidente Prudente, Brazil

    Directory of Open Access Journals (Sweden)

    Denise Cremonezi

    Full Text Available The AIDS epidemic is spreading rapidly among women worldwide, offering increasing opportunities for vertical transmission of HIV. In Brazil, the prevalence of HIV infection among pregnant women is less than 1%. Therefore, the positive predictive value of an HIV EIA test tends to be lower than the more frequent indeterminate Western blot result. Pregnant women receiving antenatal care, from 2000 to 2004, at a public secondary hospital in the city of Presidente Prudente, São Paulo, Brazil, were systematically screened for HIV by means of two distinct EIA tests, in order to determine the prevalence of indeterminate Western blot results among pregnant women showing discordance in both HIV EIA tests and indirect immunofluorescence assay. Confirmatory indirect immunofluorescence was performed on material for all women with positive results in both EIA tests. Whenever there were positive results in EIA and IIA, the applicant was retested by the initial screening assay. Only those not showing concordance in results in EIA and IAA had a Western blot performed. The viral load was measured in pregnant women with positive or indeterminate Western blot results. Out of 9,786 sera, 105 (1.0% were positive in the two HIV EIA screening tests, confirmed by indirect immunofluorescence. Among these women, Western blot was interpreted as indeterminate in 11 (0.1% cases and their viral load was <50 copies/mL. We found a prevalence of 0.1% HIV indeterminate Western blots in pregnant women from Presidente Prudente and the surrounding region; none of these pregnant women had positive HIV viral loads.

  10. 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

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

    Science.gov (United States)

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

    2010-07-14

    Avian Borna virus (ABV) has recently been shown to be the causal agent of proventricular dilatation disease (PDD) a lethal neurologic disease of captive psittacines and other birds. An immunoblot assay was used to detect the presence of antibodies against avian Borna virus in the serum of affected birds. A lysate from ABV-infected duck embryo fibroblasts served as a source of antigen. The assay was used to test for the presence of antibodies to ABV in 117 birds. Thirty of these birds had biopsy or necropsy-confirmed proventricular dilatation disease (PDD), while the remaining 87 birds were apparently healthy or were suffering from diseases other than PDD. Sera from 27 of the 30 PDD cases (90%) contained antibodies to ABV. Seventy-three (84%) of the apparently "healthy" birds were seronegative. Additionally, sera from seven macaws and one parrot trapped in the Peruvian Amazon were seronegative. Positive sera recognized the bornaviral nucleoprotein (N-protein). While the presence of antibodies to ABV largely corresponded with the development of clinical PDD, 14 apparently healthy normal birds possessed detectable antibodies to ABV. The existence of a carrier state was confirmed when 13 of 15 apparently healthy cockatiels were shown by PCR to have detectable ABV RNA in their feces. Western blot assays may be of significant assistance in diagnosing proventricular dilatation disease. Many apparently healthy birds may however be seronegative while, at the same time, shedding ABV in their feces. (c) 2009 Elsevier B.V. All rights reserved.

  12. Evaluation of Western blot, ELISA and latex agglutination tests to detect Toxoplasma gondii serum antibodies in farmed red deer.

    Science.gov (United States)

    Patel, Kandarp Khodidas; Howe, Laryssa; Heuer, Cord; Asher, Geoffery William; Wilson, Peter Raymond

    2017-09-15

    Abortion due to Toxoplasma gondii has been suspected in New Zealand farmed red deer. However, knowledge around the epidemiology and prevalence of T. gondii in farmed red deer is limited. The aim of this study was to firstly, assess the sensitivity and specificity of two commercially available assays, ELISA and latex agglutination test (LAT), for use in deer and secondly, to estimate the sero-prevalence of T. gondii in red deer. A total of 252 sera from rising 2-year-old and adult hinds from 17 New Zealand red deer herds at early and late pregnancy scanning and from known aborted and/or non-aborted hinds were tested for the presence of T. gondii antibodies. Each assays' sensitivity and specificity was evaluated by both the Western Blot (WB) as a gold standard method and Bayesian latent class (BLC) analysis in the absence of a gold standard. The sensitivity and specificity for WB were 95.8% (95% credible interval: 89.5-99.2%) and 95.1% (95% credible interval: 90.6-98.1%), respectively. For the LAT at the manufacturer's recommended ≥1:32 cut-off titre, the sensitivity (88.7%, 95% credible interval: 80.8-94.7%) and specificity (74.3%, 95% credible interval: 67.5-80.5%) were lower and higher than the sensitivity (76.2%, 95% credible interval: 66.7-84.5%) and specificity (89.7%, 95% credible interval: 84.5-93.9%) at a ≥1:64 cut-off, using (BLC) analysis. Sensitivity and specificity of the LAT at cut-off titre of 1:32 were estimated to be 84.4% (95% CI: 74.9-90.9%) and 73.5% (95% CI: 65.8-79.9%) against WB. The LAT had better agreement with WB at cut-off titre of ≥1:64 than ≥1:32 (Kappa=0.63 vs 0.54). At optimised cut-off S/P of 15.5%, the sensitivity (98.8%, 95% credible interval 96.1-99.8%) and specificity (92.8%, 95% credible interval 88.9-95.7%) of the ELISA were higher and lower, respectively, than the sensitivity (85.1%, 95% credible interval 76.2-91.9%) and specificity (98.5%, 95% credible interval 96.9-99.4%) at manufacturer's cut-off S/P of 30%, from BLC

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

    Science.gov (United States)

    Turillazzi, Emanuela; Di Paolo, Marco; Neri, Margherita; Riezzo, Irene; Fineschi, Vittorio

    2014-07-02

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

  14. 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.

  15. Proviral Features of Human T Cell Leukemia Virus Type 1 in Carriers with Indeterminate Western Blot Analysis Results.

    Science.gov (United States)

    Kuramitsu, Madoka; Sekizuka, Tsuyoshi; Yamochi, Tadanori; Firouzi, Sanaz; Sato, Tomoo; Umeki, Kazumi; Sasaki, Daisuke; Hasegawa, Hiroo; Kubota, Ryuji; Sobata, Rieko; Matsumoto, Chieko; Kaneko, Noriaki; Momose, Haruka; Araki, Kumiko; Saito, Masumichi; Nosaka, Kisato; Utsunomiya, Atae; Koh, Ki-Ryang; Ogata, Masao; Uchimaru, Kaoru; Iwanaga, Masako; Sagara, Yasuko; Yamano, Yoshihisa; Okayama, Akihiko; Miura, Kiyonori; Satake, Masahiro; Saito, Shigeru; Itabashi, Kazuo; Yamaguchi, Kazunari; Kuroda, Makoto; Watanabe, Toshiki; Okuma, Kazu; Hamaguchi, Isao

    2017-09-01

    Western blotting (WB) for human T cell leukemia virus type 1 (HTLV-1) is performed to confirm anti-HTLV-1 antibodies detected at the initial screening of blood donors and in pregnant women. However, the frequent occurrence of indeterminate results is a problem with this test. We therefore assessed the cause of indeterminate WB results by analyzing HTLV-1 provirus genomic sequences. A quantitative PCR assay measuring HTLV-1 provirus in WB-indeterminate samples revealed that the median proviral load was approximately 100-fold lower than that of WB-positive samples (0.01 versus 0.71 copy/100 cells). Phylogenic analysis of the complete HTLV-1 genomes of WB-indeterminate samples did not identify any specific phylogenetic groups. When we analyzed the nucleotide changes in 19 HTLV-1 isolates from WB-indeterminate samples, we identified 135 single nucleotide substitutions, composed of four types, G to A (29%), C to T (19%), T to C (19%), and A to G (16%). In the most frequent G-to-A substitution, 64% occurred at GG dinucleotides, indicating that APOBEC3G is responsible for mutagenesis in WB-indeterminate samples. Moreover, interestingly, five WB-indeterminate isolates had nonsense mutations in Pol and/or Tax, Env, p12, and p30. These findings suggest that WB-indeterminate carriers have low production of viral antigens because of a combination of a low proviral load and mutations in the provirus, which may interfere with host recognition of HTLV-1 antigens. Copyright © 2017 American Society for Microbiology.

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

    Science.gov (United States)

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

  17. Blood donors with indeterminate anti-p24gag reactivity in HIV-1 western blot: absence of infectivity to transfused patients and in virus culture

    NARCIS (Netherlands)

    van der Poel, C. L.; Lelie, P. N.; Reesink, H. W.; van Exel-Oehlers, P. J.; Tersmette, M.; van den Akker, R.; Gonzalves, M.; Huisman, J. G.

    1989-01-01

    During a follow-up period of 23-40 months, 7 regular blood donors had persistently, and 4 had intermittently indeterminate anti-p24gag reactivity in human immunodeficiency virus (HIV)-1 Western Blot. Serological testing and viral cultures revealed that these donors had no signs of infection for

  18. Occurrence, characteristics, and patterns of HIV-1 and HIV-2 western blot indeterminate sera in low risk populations in West Virginia and pre-AIDS Africa.

    Science.gov (United States)

    Schindzielorz, A H; Belshe, R B; Mufson, M A

    1990-05-01

    To further characterize HIV-1 and HIV-2 Western blot indeterminate (IWB) sera, 402 sera from 318 healthy low-risk individuals from West Virginia and 159 African sera obtained in the pre-AIDS era (1968-1972) were studied. All IWB sera tested for antigen by HIV-1 enzyme immunoassay (EIA-Ag) were negative. HIV-1 and HIV-2 IWB reactivity occurred independent of HIV-1 and HIV-2 false-positive testing for antibody by enzyme immunoassay (EIA-Ab) and no cross-reactions between HIV-1 and HIV-2 IWB patterns were detected. The IWB patterns were reproducible, demonstrated no age or sex related pattern, and showed no evidence of vertical or horizontal transmission. The African sera exhibited a significantly higher number of IWB patterns. IWB reactivity in HIV-1 and HIV-2 seronegative individuals may not be viral in origin and the occurrence of IWB pattern may vary among populations.

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

    Directory of Open Access Journals (Sweden)

    Hong-yuan DUAN

    2011-03-01

    Full Text Available 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 a recombinant plasmid.With E.coli BL21(DE3,the recombinant protein was induced for expression.After purification,the Western-blot assay was applied as a method to evaluate immune-reaction between rLipL32 and pathogenic Leptospira antibodies of 15 serogroups including Leptospira icterohaemorrhagiae strain 56601.Results As the result,the recombinant protein rLipL32 coded by plasmid pET28a-LipL32 was expressed as an inclusion-body.The immune-reaction with Western blotting test truly occurred between rLipL32 and antibodies against 15 serogroups of Leptospira.Conclusions In conclusion,the expressed outer membrane protein rLipL32 do show cross-reaction among members of 15 serogroups of pathogenic Leptospira.It is possible to choose rLipL32 as the antigen for antibody detection in serodiagnosis of pathogenic Leptospira within genus.

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

    Science.gov (United States)

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

    2013-12-01

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

  1. Expansion of HIV screening to non-clinical venues is aided by the use of dried blood spots for Western blot confirmation.

    Science.gov (United States)

    Sullivan, Timothy J; Antonio-Gaddy, Mara San; Richardson-Moore, April; Styer, Linda M; Bigelow-Saulsbery, Deborah; Parker, Monica M

    2013-12-01

    HIV rapid testing programs in New York State (NYS) are required to collect a specimen for confirmation of a preliminary positive result; however, some venues have limited capacity to collect venous blood, and confirmation using oral fluid is restricted by cost and availability. To evaluate the feasibility of using dried blood spots (DBS) at non-clinical HIV rapid testing sites for Western blot testing. The New York State Department of Health facilitated registration of 48 non-clinical HIV test sites and provided training on DBS procedures. Following a reactive rapid test, DBS were collected by fingerstick onto filter paper cards, dried and mailed to the NYS public health laboratory for Western blot testing. From October 2010 to December 2012, 280 DBS specimens were submitted for confirmation. Four (1.4%) were unsatisfactory for testing and 276 (98.6%) DBS were tested. Of these, 235 (85.1%) were positive, 37 (13.4%) were negative and 4 (1.4%) were indeterminate. During this period, the laboratory also received 1033 venous blood specimens for rapid test confirmation, and 35 (3.4%) were unsatisfactory. Of the 998 tested by Western blot, 784 (78.6%) were positive, 197 (19.7%) were negative and 17 (1.7%) were indeterminate. Compared to venous blood, the percentage of rapid test referral specimens with a positive Western blot was significantly greater for DBS specimens and the frequency of unsatisfactory specimens did not differ significantly. These results indicate that DBS are a suitable alternative to venous blood for confirmation of HIV rapid tests conducted at non-clinical sites. Copyright © 2013 Elsevier B.V. All rights reserved.

  2. Anti-RAINBOW dye-specific antibodies as universal tools for the visualization of prestained protein molecular weight markers in Western blot analysis.

    Science.gov (United States)

    Schüchner, Stefan; Andorfer, Peter; Mudrak, Ingrid; Ogris, Egon

    2016-08-17

    Western blotting is one of the most widely used techniques in molecular biology and biochemistry. Prestained proteins are used as molecular weight standards in protein electrophoresis. In the chemiluminescent Western blot analysis, however, these colored protein markers are invisible leaving researchers with the unsatisfying situation that the signal for the protein of interest and the signal for the markers are not captured simultaneously and have to be merged in an error-prone step. To allow the simultaneous detection of marker proteins we generated monoclonal antibodies specific for the protein dyes. To elicit a dye rather than protein specific immune response we immunized mice sequentially with dye-carrier protein complexes, in which a new carrier protein was used for each subsequent immunization. Moreover, by sequentially immunizing with dye-carrier protein complexes, in which different but structurally related dyes were used, we could also generate an antibody, termed anti-RAINBOW, that cross-reacted even with structurally related dyes not used in the immunizations. Our novel antibodies represent convenient tools for the simultaneous Western blot detection of commercially available prestained marker proteins in combination with the detection of any specific protein of interest. These antibodies will render obsolete the anachronistic tradition of manually charting marker bands on film.

  3. Re-purposing of histological tissue sections for corroborative western blot analysis of hypothalamic metabolic neuropeptide expression following delineation of transactivated structures by Fos immuno-mapping.

    Science.gov (United States)

    Alenazi, Fahaad S H; Ibrahim, Baher A; Briski, Karen P

    2015-04-01

    Fos immunocytochemistry is a valuable anatomical mapping tool for distinguishing cells within complex tissues that undergo genomic activation, but it is seldom paired with corroborative molecular analytical techniques. Due to preparatory requirements that include protein cross-linking for specimen sectioning, histological tissue sections are regarded as unsuitable for those methods. Our studies show that pharmacological activation of the hindbrain energy sensor AMPK by AICAR elicits estradiol (E)-dependent patterns of Fos immunolabeling of hypothalamic metabolic loci. Here, Western blotting was applied to hypothalamic tissue removed from histological sections of E- versus oil (O)-implanted ovariectomized (OVX) female rat brain to measure levels of metabolic transmitters associated with Fos-positive structures. In both E and O rats, AICAR treatment elicited alterations in pro-opiomelanocortin, neuropeptide Y, SF-1, and orexin-A neuropeptide expression that coincided with patterns of Fos labeling of structures containing neurons that synthesize these neurotransmitters, e.g. arcuate and ventromedial nuclei and lateral hypothalamic area. O, but not E animals also exhibited parallel augmentation of tissue corticotropin-releasing hormone neuropeptide levels and paraventricular nucleus Fos staining. Data demonstrate the utility of immunoblot analysis as a follow-through technique to capitalize on Fos mapping of transactivation sites in the brain. Findings that induction of Fos immunoreactivity coincides with adjustments in hypothalamic metabolic neuropeptide expression affirms that this functional indicator reflects changes in neurotransmission in pathways governing metabolic outflow. Copyright © 2015 Elsevier Ltd. All rights reserved.

  4. Immunohistochemical and western blot analysis suggest that the soluble forms of FGF1-2 and FGFR1-2 sustain tail regeneration in the lizard.

    Science.gov (United States)

    Alibardi, Lorenzo

    2017-11-01

    Fibroblast Growth Factors 1-2 (FGF1-2) stimulate tail regeneration in lizards and therefore the distribution of their receptors, FGFR1-2, in the regenerating tail of the lizard. Podarcis muralis has been studied using immunofluorescence and western blotting. Immunoreactive protein bands at 15-16kDa for FGF1-2 in addition to those at 50-65kDa are detected in the regenerating epidermis, but weak bands at 35, 45 and 50kDa appear from the regenerating connective tissues. Strongly immunolabeled bands for FGFR1 at 32, 60, and 80kDa and less intense for FGFR2 only appear in the regenerating tail. In normal tail epidermis and dermis, higher MW forms are present at 80 and 115-140kDa, respectively, but they disappear in the regenerating epidermis and dermis where low MW forms of FGFR1-2 are found at 50-70kDa. Immunolocalization confirms that most FGFR1-2 are present in the wound epidermis, Apical Epidermal Peg, ependymal tube while immunolabeling lowers in regenerating muscles, blastema cells, cartilage and connectives tissues. The likely release of FGFs from the Apical Epidermal Peg and ependyma and the presence of their receptors in these tissues may determine the autocrine stimulation of proliferation and a paracrine stimulation of the blastema cells through their FGF Receptors. Copyright © 2017 Elsevier GmbH. All rights reserved.

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

    Science.gov (United States)

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

    2014-08-27

    Zoonotic transmission of hepatitis E virus (HEV) is of special concern, particularly in high income countries were waterborne infections are less frequent than in developing countries. High HEV seroprevalences can be found in European pig populations. The aims of this study were to obtain prevalence data on HEV infection in swine in Belgium and to phylogenetically compare Belgian human HEV sequences with those obtained from swine. An ELISA screening prevalence of 73% (95% CI 68.8-77.5) was determined in Belgian pigs and a part of the results were re-evaluated by Western blot (WB). A receiver operating characteristic curve analysis was performed and scenarios varying the ELISA specificity relative to WB were analysed. The seroprevalences estimated by the different scenarios ranged between 69 and 81% and are in agreement with the high exposure of the European pig population to HEV. Pig HEV sequences were genetically compared to those detected in humans in Belgium and a predominance of genotype 3 subtype f was shown in both swine and humans. The high HEV seroprevalence in swine and the close phylogenetic relationships between pig and human HEV sequences further support the risk for zoonotic transmission of HEV between humans and pigs. Copyright © 2014 Elsevier B.V. All rights reserved.

  6. [Western Blot analysis of type I, III, V, VI collagen after laser epithelial keratomileusis and photorefractive keratectomy in cornea of rabbits].

    Science.gov (United States)

    Cui, Xin; Bai, Ji; He, Xiangge; Zhang, Yi

    2005-12-01

    Use immunohistochemical staining and Western Blot analysis to observe and compare the accurate dynamic changes of type I, III, V, VI collagen in the wound healing processes of the rabbit cornea which underwent LASEK or PRK to investigate the possible mechanism of corneal haze and myopic regression. New Zealand White rabbits were divided into 8 groups: normal control group (n=6), 1 day, 7 days, 1, 3, 4, 5 and 6 month groups (n=14). Every rabbit underwent LASEK in one eye while the other one with PRK. We use immunohistochemical staining and Western Blot analysis to compare the wound healing process of dynamic change of the type I, III, V and VI collagen in rabbit cornea of every time point. The results were analysised with data analysis software. Immunohistochemical staining and Western Blot analysis showed that after LASEK, the cornea wound healing with type I and III collagen were much faster than PRK, and the wound response was also much weaker. Whereas for type V and VI collagen, their dynamic changes were resemble between LASEK and PRK, they both reached the peak value after 3 months since the surgery, but LASEK group returned to normal earlier than PRK. The value of these two types of collagen after PRK were higher than LASEK. The changes of these four types of collagen may offer us at least partial explaination to the difference between formation between corneal haze and refractive regression. There were significant differences between LASEK and PRK on type I, III, V and VI collagens or the time of reacting, reaching apex and returning to normal . LASEK had slighter intensity of reaction. The results indicate that there is excessive aggradation of collagens after PRK, it may be the histological foundation of obvious haze and myopia regression.

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

    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.

  8. Accuracy of indirect haemagglutination and western blot assays for the detection of anti-Schistosoma antibodies in non-severe febrile patients in two Tanzanian hospitals.

    Science.gov (United States)

    Bevilacqua, Nazario; Pane, Stefania; Vairo, Francesco; Nicastri, Emanuele; Paglia, Maria G; Ame, Shaali M; Schepisi, Monica Sañé; Kitua, Andrew; Mangi, Sabina; Racalbuto, Vincenzo; Meschi, Silvia; Ippolito, Giuseppe

    2012-06-01

    The diagnosis of schistosomiasis is usually based on clinical data associated with the detection of eggs in stool, urine, and/or rectal and bladder biopsy specimens. However antibody detection can be useful to indicate Schistosoma infection in those for whom eggs cannot be demonstrated. The aim of this study was to assess the seroprevalence of schistosomiasis and to evaluate the accuracy of indirect haemagglutination (IHA) and Western blot (WB) assays for the detection of anti-Schistosoma antibodies in 2 peripheral hospitals of the United Republic of Tanzania. Between February and March 2007 blood samples were collected from 297 non-severe febrile outpatients who attended Chake Chake Hospital, Pemba Island and Tosamaganga Hospital, Iringa region in Tanzania. The samples were processed for Schistosoma antibodies by IHA and WB assays in Italy. Two hundred and sixty-two of 297 patients were schistosomiasis antibody-positive by IHA (88.2%). Of 142 patients positive by IHA, only 22 (12.4%) cases were confirmed by WB assay. The WB assay confirmed all 35 negative cases previously identified by IHA. The seroprevalence of Schistosoma at Chake Chake Hospital was lower than in Tosamaganga Hospital (9/97, 9.3% vs 13/80, 16.2%). Schistosomiasis is endemic in Tanzania, being more prevalent on the mainland than on Pemba Island. The implications of this study are of public health relevance and suggest the need for increased efforts in large-scale chemotherapy-based morbidity control programmes, integrated with those for other soil-transmitted helminthiases, in these 2 peripheral areas of the United Republic of Tanzania.

  9. Carbonylation and glutathionylation of proteins in the blue mussel Mytilus edulis detected by proteomic analysis and Western blotting: Actin as a target for oxidative stress.

    Science.gov (United States)

    McDonagh, Brian; Tyther, Raymond; Sheehan, David

    2005-07-01

    Protein expression profiles (PEPs) were generated by two-dimensional electrophoresis (2-D SDS-PAGE) for gill and digestive glands of Mytilus edulis sampled from a polluted and reference site in Cork Harbour, Ireland. Similar patterns and expression levels were found for both sites in silver stained gels. However, Western blotting for carbonylated proteins demonstrated higher levels of specific carbonylation of proteins in tissues from animals in the polluted site. Animals from the reference site were acclimated in holding tanks, exposed to 1 mM H2O2 for 24 h, dissected and analysed by 2-D SDS-PAGE. Again, generally similar PEPs were found in control and exposed animals for gill and digestive gland but carbonylation was more pronounced in polluted and exposed animals. Western blotting of extracts after one-dimensional electrophoresis with antibodies to glutathione and actin revealed that gill proteins are glutathionylated more strongly than digestive gland and that this process is more pronounced in polluted animals than in controls. We conclude that carbonylation and glutathionylation can occur in gill and digestive gland in response to oxidative stress in M. edulis. Actin is a major target for both glutathionylation and carbonylation under oxidative stress conditions.

  10. Evaluation of Humoral Immune Response against Somatic and Excretory-Secretory Antigens of Dicrocoelium Dendriticum in Infected Sheep by Western Blot

    Directory of Open Access Journals (Sweden)

    Mohammad Hosein Razi Jalali

    2013-12-01

    Full Text Available Introduction and objective: Dicrocoelium dendriticum is a worldwide spread parasite of liver, bile ducts and gallbladder of especially ruminants and humans as well. Identification of specific antigens is useful for early diagnosis of the infection. The goal of this study was the isolation and identification of excretory-secretory and somatic antigens from D. dendriticum by sodium dodecyl sulphate (SDS-PAGE and evaluation of humoral immune response against these antigens.   Methods: The parasites were collected and washed by phosphate buffered saline (PBS and supplemented by antibiotic for several times. For preparing somatic antigens, parasites were sonicated and centrifuged prior to collect supernatant. For preparing excretory-secretory antigens the viable parasites were transferred to the sterile medium. The samples were centrifuged and supernatants were collected. The sera of infected sheep with different infection degrees were collected too. Somatic and excretory-secretory proteins were isolated with SDS PAGE and stained with coomassie blue. Immunogenicity properties of the resulting proteins were determined using western blot analysis.   Results: The total extract of somatic antigens analyzed by SDS-PAGE revealed 21 proteins. In mild infection, bands of 130 KDa were immune dominant. In moderate infections 48, 80 and 130 KDa and in heavy infections 48, 60, 80, 130 KDa were detected as immune dominant bands. In excretory- secretory antigens seven bands of protein were detected. In mild infection 130 KDa, in moderate infection 100, 120 and 130 KDa and in heavy infection 45, 80, 85, 100, 120 and 130 KDa were immune dominant bands.   Conclusion: Probably the most immunogenic protein band during different degrees of infection was 130KDa that can be used for vaccination and inducing immunity.

  11. 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

    2018-01-10

    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.

  12. 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

    .... Common Western blotting techniques were used to compare immunostaining patterns of tissue lysates between a known species, rat, and the guinea pig using antibodies to several common CNS proteins...

  13. A comparison of antigenic peptides in muscle larvae of several Trichinella species by two-dimensional western-blot analysis with monoclonal antibodies

    Directory of Open Access Journals (Sweden)

    Dea-Ayuela M.A.

    2001-06-01

    Full Text Available The antigens recognised by mAb US5 specific to 53 kDa glycoprotein (gp 53 in T. spiralis L-1 muscle larvae (TSL1 antigens, mAb US9 specific to gp 53 in TSL1 from all encapsulated species and mAb US4 specific to a tyvelose containing tetrasaccharide present in TSL1, were investigated in crude extracts from muscle larvae of T. spiralis, T. nativa and T. britovi by 2D-electrophoresis and western-blot. At least four proteins of different pI were recognised by mAb US5 on T. spiralis antigens. Recognition profile of mAb US9 on T. spiralis antigens exhibited some variation with regard to that of the US5. Polymorphism was apparent in gp 53. High reactivity was shown by the mAb US4 with the three species.

  14. First detection of CYP1A1 and CYP2B induction in Mediterranean cetacean skin biopsies and cultured fibroblasts by Western blot analysis.

    Science.gov (United States)

    Fossi, Maria Cristina; Casini, Silvia; Bucalossi, Daniela; Marsili, Letizia

    2008-07-01

    The need to develop powerful tools to detect exposure and effects of POPs and emerging contaminants in Mediterranean cetaceans led us to develop a suite of sensitive non-lethal biomarkers in integument biopsies of free-ranging animals. In order to propose induction of CYP1A1 and CYP2B, detected by Western blot analysis, as biomarkers of exposure to OCs, PAHs and PBDEs, a three-phase experimental protocol (in vitro experiments, calibration experiments and field applications) was followed using fibroblast cell cultures and biopsies of Mediterranean Stenella coeruleoalba and Tursiops truncatus. This methodology was confirmed to be sensitive and stable in comparison to previous methods used to detect CYP1A1 in biopsies, enabling analysis of several inducible proteins in non-lethal samples and analysis of material from stranded animals.

  15. Use of rapid HIV assays as supplemental tests in specimens with repeatedly reactive screening immunoassay results not confirmed by HIV-1 Western blot.

    Science.gov (United States)

    Wesolowski, Laura G; Delaney, Kevin P; Meyer, William A; Blatt, Amy J; Bennett, Berry; Chavez, Pollyanna; Granade, Timothy C; Owen, Michele

    2013-09-01

    An alternate HIV testing algorithm has been proposed which includes a fourth-generation immunoassay followed by an HIV-1/HIV-2 antibody differentiation supplemental test for reactive specimens and a nucleic acid test (NAT) for specimens with discordant results. To evaluate the performance of five rapid tests (Alere Clearview, Bio-Rad Multispot, OraSure OraQuick, MedMira Reveal, and Trinity Biotech Unigold) as the supplemental antibody assay in the algorithm. A total of 3273 serum and plasma specimens that were third-generation immunoassay repeatedly reactive and Western blot (WB) negative or indeterminate were tested with rapid tests and NAT. Specimens were classified by NAT: (1) HIV-1 infected (NAT-reactive; n=184, 5.6%), (2) HIV-status unknown (NAT nonreactive; n=3078, 94.2%) or by Multispot, (3) HIV-2 positive (n=5), and (4) HIV-1 and HIV-2 positive (n=6). Excluding HIV-2 positive specimens, we calculated the proportion of reactive rapid tests among specimens with reactive and nonreactive NAT. The proportion of infected specimens with reactive rapid test results and negative or indeterminate WB ranged from 30.4% (56) to 47.8% (88) depending on the rapid test. From 1% to 2% of NAT-negative specimens had reactive rapid test results. In these diagnostically challenging specimens, all rapid tests identified infections that were missed by the Western blot, but only Multispot could differentiate HIV-1 from HIV-2. Regardless of which rapid test is used as a supplemental test in the alternative algorithm, false-positive algorithm results (i.e., reactive screening and rapid test in uninfected person) may occur, which will need to be resolved during the baseline medical evaluation. Published by Elsevier B.V.

  16. 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.

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

    Science.gov (United States)

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

    2016-12-23

    Sera of 89 pregnant women were selected according to the results of ELFA IgM, IgG and avidity IgG, and tested with commercial tests IgM, IgG and avidity IgG Western Blot (WB) to compare the efficacy of both techniques in determining the phase of T. gondii infection. In total, 81 of 89 tested sera (91.0%) were classified as positive, both in the ELFA and WB tests for the presence of anti-Toxoplasma antibodies of class IgG, indicating a past infection, while the prevalence of anti-Toxoplasma positive reactions associated with the antibodies of class IgM indicating a recent infection was much lower - 31.5% and 20.2%, respectively. Sera of 81 women were also tested in the ELFA and WB tests for avidity, e.g. ability of forming high-molecular IgG antibody complexes. Low or medium results in these tests (in this study all classified as low), indicating a recent infection, were detected by ELFA and WB in 22.2% and 45.7% of the total examined samples, respectively. The Spearman's rank test for correlation, performed for recognition of quantitative data of the ELFA and WB tests (index, units or points), revealed a highly significant correlation between the ELFA and WB tests for homologous classes of antibodies, both for IgM and IgG (p0.05), except for the WB test for IgM antibodies, which showed a significant correlation with the ELFA test for IgG antibodies (p<0.01). A highly significant negative correlation between the ELFA and WB test for IgM antibodies and ELFA and WB tests for IgG avidity was demonstrated (p<0.01), except for a relationship between the WB test for IgM and WB for avidity, which was not significant. Such negative correlations are theoretically expected, as strong complexes with the participation of IgG antibodies are absent in the early phase of toxoplasmosis when early antibodies of IgM class are present. Summarizing, this study indicates the high usefulness of the commercial ELFA and WB tests in serodiagnostics of toxoplasmosis in pregnant women

  18. Diagnostic performance of ELISA, IFAT and Western blot for the detection of anti-Leishmania infantum antibodies in cats using a Bayesian analysis without a gold standard.

    Science.gov (United States)

    Persichetti, Maria Flaminia; Solano-Gallego, Laia; Vullo, Angela; Masucci, Marisa; Marty, Pierre; Delaunay, Pascal; Vitale, Fabrizio; Pennisi, Maria Grazia

    2017-03-13

    Anti-Leishmania antibodies are increasingly investigated in cats for epidemiological studies or for the diagnosis of clinical feline leishmaniosis. The immunofluorescent antibody test (IFAT), the enzyme-linked immunosorbent assay (ELISA) and western blot (WB) are the serological tests more frequently used. The aim of the present study was to assess diagnostic performance of IFAT, ELISA and WB to detect anti-L. infantum antibodies in feline serum samples obtained from endemic (n = 76) and non-endemic (n = 64) areas and from cats affected by feline leishmaniosis (n = 21) by a Bayesian approach without a gold standard. Cut-offs were set at 80 titre for IFAT and 40 ELISA units for ELISA. WB was considered positive in presence of at least a 18 KDa band. Statistical analysis was performed through a written routine with MATLAB software in the Bayesian framework. The latent data and observations from the joint posterior were simulated in the Bayesian approach by an iterative Markov Chain Monte Carlo technique using the Gibbs sampler for estimating sensitivity and specificity of the three tests. The median seroprevalence in the sample used for evaluating the performance of tests was estimated at 0.27 [credible interval (CI) = 0.20-0.34]. The median sensitivity of the three different methods was 0.97 (CI: 0.86-1.00), 0.75 (CI: 0.61-0.87) and 0.70 (CI: 0.56-0.83) for WB, IFAT and ELISA, respectively. Median specificity reached 0.99 (CI: 0.96-1.00) with WB, 0.97 (CI: 0.93-0.99) with IFAT and 0.98 (CI: 0.94-1.00) with ELISA. IFAT was more sensitive than ELISA (75 vs 70%) for the detection of subclinical infection while ELISA was better for diagnosing clinical leishmaniosis when compared with IFAT (98 vs 97%). The overall performance of all serological techniques was good and the most accurate test for anti-Leishmania antibody detection in feline serum samples was WB.

  19. Detecting Allergens From Black Tiger Shrimp Penaeus monodon That Can Bind and Cross-link IgE by ELISA, Western Blot, and a Humanized Rat Basophilic Leukemia Reporter Cell Line RS-ATL8.

    Science.gov (United States)

    Jarupalee, Thanyapat; Chatchatee, Pantipa; Komolpis, Kittinan; Suratannon, Narissara; Roytrakul, Sittiruk; Yingchutrakul, Yodying; Yimchuen, Wanaporn; Butta, Patcharavadee; Jacquet, Alain; Palaga, Tanapat

    2018-01-01

    Black tiger shrimp Penaeus monodon is one of the common causes of shellfish allergy that is increasing worldwide. One of the important problems in the management of shellfish allergy is the lack of accurate diagnostic assay because the biological and immunological properties of allergens in black tiger shrimp have not been well characterized. This study aims to detect proteins with the ability to bind and cross-link immunoglobulin E (IgE) from black tiger shrimp by enzyme-linked immunosorbent assay (ELISA), Western blot, and a humanized rat basophilic leukemia reporter cell line RS-ATL8. Sera from shrimp allergic subjects were subjected to ELISA and Western blots using raw or cooked shrimp extract as antigens. Pooled sera were used to sensitize the RS-ATL8 reporter cell line and cells were activated by shrimp extract. Eluted protein extracts separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) were tested on the RS-ATL8 cell line and subjected to mass spectrometry to identify potential candidate allergens. Allergic sera reacted stronger to raw shrimp extract than cooked shrimp extract (P=0.009). Western blot demonstrated that major IgE reactivity protein bands were at 32-39 kDa and 91-230 kDa in both raw and cooked shrimp extracts. The eluted protein bands at the molecular weight of 38 and 115 kDa from raw shrimp extract induced IgE cross-linking as assayed by the RS-ATL8 cell line. These protein bands were subjected to mass spectrometry for analysis. Ubiquitin-activating enzyme and crustacyanin were identified as potential candidate novel shrimp allergens. The RS-ATL8 reporter cell line can be used to identify potential new shrimp allergens that can functionally cross-link IgE and induce mast cell degranulation.

  20. Bicentric evaluation of six anti-toxoplasma immunoglobulin G (IgG) automated immunoassays and comparison to the Toxo II IgG Western blot.

    Science.gov (United States)

    Maudry, Arnaud; Chene, Gautier; Chatelain, Rémi; Patural, Hugues; Bellete, Bahrie; Tisseur, Bernard; Hafid, Jamal; Raberin, Hélène; Beretta, Sophie; Sung, Roger Tran Manh; Belot, Georges; Flori, Pierre

    2009-09-01

    A comparative study of the Toxoplasma IgG(I) and IgG(II) Access (Access I and II, respectively; Beckman Coulter Inc.), AxSYM Toxo IgG (AxSYM; Abbott Diagnostics), Vidas Toxo IgG (Vidas; bioMerieux, Marcy l'Etoile, France), Immulite Toxo IgG (Immulite; Siemens Healthcare Diagnostics Inc.), and Modular Toxo IgG (Modular; Roche Diagnostics, Basel, Switzerland) tests was done with 406 consecutive serum samples. The Toxo II IgG Western blot (LDBio, Lyon, France) was used as a reference technique in the case of intertechnique discordance. Of the 406 serum samples tested, the results for 35 were discordant by the different techniques. Using the 175 serum samples with positive results, we evaluated the standardization of the titrations obtained (in IU/ml); the medians (second quartiles) obtained were 9.1 IU/ml for the AxSYM test, 21 IU/ml for the Access I test, 25.7 IU/ml for the Access II test, 32 IU/ml for the Vidas test, 34.6 IU/ml for the Immulite test, and 248 IU/ml for the Modular test. For all the immunoassays tested, the following relative sensitivity and specificity values were found: 89.7 to 100% for the Access II test, 89.7 to 99.6% for the Immulite test, 90.2 to 99.6% for the AxSYM test, 91.4 to 99.6% for the Vidas test, 94.8 to 99.6% for the Access I test, and 98.3 to 98.7% for the Modular test. Among the 406 serum samples, we did not find any false-positive values by two different tests for the same serum sample. Except for the Modular test, which prioritized sensitivity, it appears that the positive cutoff values suggested by the pharmaceutical companies are very high (either for economical or for safety reasons). This led to imperfect sensitivity, a large number of unnecessary serological follow-ups of pregnant women, and difficulty in determining the serological status of immunosuppressed individuals.

  1. Evaluation of Line Immunoassay to Detect HTLV-1 Infection in an Endemic Area, Southwestern Japan; Comparison with Polymerase Chain Reaction and Western Blot.

    Science.gov (United States)

    Umeki, Kazumi; Umekita, Kunihiko; Hashikura, Yuuki; Yamamoto, Ikuo; Kubo, Kazuyoshi; Nagatomo, Yasuhiro; Okayama, Akihiko

    2017-02-01

    Human T-lymphotropic virus type 1 (HTLV-1) has been recognized as a cause of adult T-cell leukemia/lymphoma, HTLV-1-associated myelopathy/tropical spastic paraparesis, and HTLV-1-associated uveitis. HTLV-1 infection is normally detected by screening for HTLV-1 antibodies, and positive samples are confirmed by Western blot (WB). However, WB fails to confirm some samples that were positive for HTLV-1 antibodies on screening. Line immunoassay (LIA) is commonly used in Europe and Brazil, but not in Japan. Therefore, we evaluated the performance of LIA as a method of confirming HTLV-1 antibodies using samples in Japan. LIA was compared with polymerase chain reaction (PCR) and WB using 50 negative and 70 positive samples tested by chemiluminescent enzyme immunoassay (CLEIA) in Miyazaki, Japan, an HTLV-1 endemic area. LIA (INNO-LIA HTLVI/II Score) and WB (Problot HTLV-I) were performed according to the manufacturer's instructions. Real-time PCR for HTLV-1 pX region was performed using DNA derived from white blood cells. The samples that tested negative by real-time PCR were further tested by nested PCR. All 50 CLEIA negative samples were determined to be negative by LIA and PCR. Of the 70 positive samples, 66 tested positive by both of LIA and PCR. Three samples tested negative by LIA and PCR, and the remaining sample (PCR negative) showed non-specific staining in LIA and WB. WB showed more indeterminate results than LIA. Gp21 antibody in LIA demonstrated a high ability to discriminate between positive and negative PCR results. Furthermore, the degree of gp21 antibody reaction by LIA showed correlation with HTLV-1 proviral loads (PVLs). Our results indicate that LIA performs well in confirming HTLV-1 seropositivity by showing a low incidence of indeterminate results and good agreement with PCR using samples in Japan, although the number of samples tested was small. In addition, semi-quantitative antibody titer to gp21 correlated well with HTLV-1 PVLs. Further study

  2. [Recombinant expression of hantaan virus protein N with application of Western-blot in detecting anti-hantavirus antibody].

    Science.gov (United States)

    Yao, P P; Xu, F; Sun, Y S; Yang, Z R; Zhang, Y; Yue, M; Zhu, H P

    2017-04-10

    Objective: S gene of hantavirus(HV) was expressed in insect cells by genetic engineering technology. The expression product of S gene was used as antigen to detect anti-HV specific antibody IgG in serum. Methods: Gene encoding NP of the strain HV-Z10 was amplified by PCR and then its eukaryotic expression system rBAC-Z10S-TN was constructed by using the routine genetic engineering method. SDS-PAGE was applied to measure the expression of rNP.Ion-exchange plus Ni-NTA-affinity chromatography was performed to purify the recombinant product. Indirect immuno-fluorescence assay (IFA) was used to determine the specific immune-reactivity of rNP. WB assay was established to detect the serum samples from 95 confirmed HFRS patients. Parameters related to the outcomes of detection were compared with the routine HV-IgG IFA method. Results: rBAC-Z10S-TN was able to express rNP with high efficiency. The purified rNP only showed a single protein fragment in the gel after SDS-PAGE. HV IgG could efficiently recognize rNP and hybridize with the recombinant protein. 97.67% of the serum samples from the HFRS patients were positive confirmed by WB. Conclusions: We successfully constructed a high efficient prokaryotic expression system of NP encoding gene from hantavirus strain HV-Z10. WB assay which was established in this study could be used as a new serological test for HFRS diagnosis, thanks to the simplicity, safety, sensitivity and specificity of this method.

  3. 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.

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

    Directory of Open Access Journals (Sweden)

    Jacek Sroka

    2016-09-01

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

  5. Enzyme-linked immunosorbent assay and Western blot antibody determination in sera from patients diagnosed with different helminthic infections with Anisakis simplex antigen purified by affinity chromatography

    Directory of Open Access Journals (Sweden)

    M Rodero

    2005-05-01

    Full Text Available An evaluation of the sensitivity and the specificity of the Anisakis simplex antigens purified by affinity chromatography was performed using sera from patients diagnosed with Anisakis sensitisation and sera from patients previously diagnosed with different helminthic infections. Only the sera of the patients diagnosed with Schistosoma mansoni or Onchocerca volvulus parasitic infections were negative against the A. simplex antigen and its purified fractions (PAK antigen: A. simplex antigen purified using columns prepared with anti-A. simplex rabbit IgG and PAS antigen: PAK antigen purified using columns prepared with anti-Ascaris suum rabbit IgG. However all the sera were positive against the A. suum antigen. In all the sera from the patients diagnosed with Anisakis sensitisation, the antibody levels detected using the purified antigens (PAK and PAS antigens were lower than the observed using the A. simplex crude extract with the highest diminution in the case of the IgG. When these same sera were tested against the A. simplex crude extract by Western blot, several bands of high molecular masses were observed as well as, intense bands at 60 and/or 40 kDa. A concentration of these last proteins was observed in the PAK and the PAS antigens. When the sensitivity and the specificity determinations were performed, only seven of the 38 patients diagnosed of Anisakis sensitisation were positive, as well as, the sera from the patients diagnosed with parasitisms by Echinococcus granulosus or Fasciola hepatica.

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

    Science.gov (United States)

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

    2017-03-01

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

  7. Sensitivity and specificity of western blot testing of cerebrospinal fluid and serum for diagnosis of equine protozoal myeloencephalitis in horses with and without neurologic abnormalities.

    Science.gov (United States)

    Daft, Barbara M; Barr, Bradd C; Gardner, Ian A; Read, Deryck; Bell, William; Peyser, Karen G; Ardans, Alex; Kinde, Hailu; Morrow, Jennifer K

    2002-10-01

    To determine sensitivity and specificity of western blot testing (WBT) of CSF and serum for diagnosis of equine protozoal myeloencephalitis (EPM) in horses with and without neurologic abnormalities. Prospective investigation. 65 horses with and 169 horses without neurologic abnormalities. CSF and serum from horses submitted for necropsy were tested for Sarcocystis neurona-specific antibody with a WBT. Results of postmortem examination were used as the gold standard against which results of the WBT were compared. Sensitivity of WBT of CSF was 87% for horses with and 88% for horses without neurologic abnormalities. Specificity of WBT of CSF was 44% for horses with and 60% for horses without neurologic abnormalities. Regardless of whether horses did or did not have neurologic abnormalities, sensitivity and specificity of WBT of serum were not significantly different from values for WBT of CSF. Ninety-four horses without EPM had histologic evidence of slight CNS inflammation. The low specificity of WBT of CSF indicated that it is inappropriate to diagnose EPM on the basis of a positive test result alone because of the possibility of false-positive test results. The high sensitivity, however, means that a negative result is useful in ruling out EPM. There was no advantage in testing CSF versus serum in horses without neurologic abnormalities. Slight CNS inflammation was common in horses with and without S neurona-specific antibodies in the CSF and should not be considered an indication of CNS infection with S neurona.

  8. 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”.

  9. 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

  10. 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?

    Science.gov (United States)

    Cánepa, Camila; Salido, Jimena; Ruggieri, Matías; Fraile, Sindy; Pataccini, Gabriela; Berini, Carolina; Biglione, Mirna

    2015-10-28

    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. (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. 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). indeterminate WB results confirmed later as HTLV-1 positive may be associated with low PVL levels. Mutations in LTR and tax are described; their functional relevance remains to be determined.

  11. 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.

  12. Comparison of Western blot (immunoblot) based on recombinant-derived p41 with conventional tests for serodiagnosis of human immunodeficiency virus infections.

    Science.gov (United States)

    Hofbauer, J M; Schulz, T F; Hengster, P; Larcher, C; Zangerle, R; Kofler, H; Fritsch, P; Wachter, H; Dierich, M P

    1988-01-01

    To evaluate the performance of a serological test for human immunodeficiency virus type 1 (HIV-1) infections based on the use of a recombinant envelope gene-derived protein as the antigen, we caused expression of a 1.4-kilobase fragment of HIV.DNA that codes for the complete gp41 transmembrane protein in an Escherichia coli expression vector and used Western blots (WB; immunoblots) prepared with recombinant material (pEX-41) to detect antibodies to HIV-1. This test detected all 339 sera which were positive by a combination of conventional serodiagnostic assays and produced no false-positive results with 311 negative samples. Also no false-positive results were obtained with 20 sera from systemic lupus erythematosus patients which had high titers of cross-reactive autoantibodies. In six cases, the pEX-41 WB proved to be more sensitive than individual assays applied on their own, and in five cases it was even more sensitive than a combination of conventional assays. We tested 221 sera in both our pEX-41 WB and a commercially available recombinant enzyme immunoassay (EIA [Abbott]). The results were identical in 188 cases. A total of 27 sera containing antibodies to gp41 as demonstrated in the pEX-41 WB, as well as the Abbott recombinant EIA, had no antibodies to the recombinant core antigen as measured in the Abbott EIA. However, 25 of these sera did stain the 24-kilodalton band on a WB with purified virus. Six sera that were positive in all of the conventional confirmatory assays and reacted strongly with the pEX-41 WB did not recognize the surface antigen used in the Abbott recombinant EIA. We conclude that the use of WB prepared with recombinant-derived p41 offers a very sensitive and specific method to detect antibodies to HIV.

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

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    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.

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

    Science.gov (United States)

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

    2014-10-01

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

  15. 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

    lung infection with P aeruginosa was further investigated by correlating the a beta ab IgG subclasses with pulmonary function in patients with cystic fibrosis. METHODS: Immunoglobulin G (IgG) and IgG subclass a beta ab were investigated by western blotting and quantified by laser scanning densitometry......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....... A longitudinal study on 43 consecutive patients with cystic fibrosis who developed chronic lung infection with P aeruginosa was performed. RESULTS: IgG subclass a beta ab appeared in all patients with chronic infection with P aeruginosa. Eleven years after the onset of infection all the patients had IgG1, 79...

  16. 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

  17. 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.

  18. Protein blotting with direct blotting electrophoresis.

    Science.gov (United States)

    Beck, S

    1988-05-01

    Direct blotting electrophoresis, a method designed to be of general application for the separation and electroblotting of macromolecules, has been adapted to produce protein blots suitable for subsequent processing by standard techniques such as dye staining or immunological detection. After their separation in a very short gel the protein bands are electrophoresed out of the gel onto an immobilizing matrix. The matrix which is moved across the bottom of the gel by a conveyor belt binds these proteins with high affinity. Once the protein samples have been loaded onto the gel and electrophoresis has been started, no further intervention is needed until the blot is completed. The total expenditure of time for such a direct blot is less than 4 h for a mixture of proteins in the molecular weight range of 14-70 kDa. The staining sensitivity of directly blotted proteins is about 200 ng protein per band as revealed by India ink staining.

  19. A comparison of extracted proteins of isolates of Dermatophilus congolensis by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting.

    Science.gov (United States)

    Makinde, A A; Gyles, C L

    1999-07-01

    Antigenic diversity within a collection of 18 isolates of Dermatophilus congolensis from different Continents was examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and by Western blotting with sera from cattle with clinical dermatophilosis using whole cell extracts obtained by three methods and one extract of extracellular products of D. congolensis. One of the methods involving the release of a lysostaphin-solubilized protein (LSP) of whole cells of D. congolensis revealed a number of discrete and easily-identifiable bands in SDS-PAGE which were found suitable for characterizing protein patterns and was, therefore, subsequently used for a comparative analysis of the proteins of all the D. congolensis isolates. Six electropherotypes (ET) of D. congolensis were identified among the 18 isolates using the protein profiles based on the presence of four protein bands at Molecular weights (MW) 62, 28, 17.4 and 16.4 kDa. The ETs were found among isolates from different animal species and from different sources with ET1 consisting of three bovine and two equine isolates; ET2, two bovine and three ovine isolates; ET3, two bovine isolates; ET4, two bovine isolates; ET5, one bovine and one ovine isolates and ET6, two bovine isolates. Immunoblotting of the extracts of D. congolensis isolates with sera from cattle with clinical dermatophilosis infection demonstrated protein bands of MW ranging from 9 kDa to 188 kDa. Sera from chronic dermatophilosis infection demonstrated a 28 kDa protein which was immunodominant in the LSP extracts of all the 18 isolates of D. congolensis tested while sera from mild infections demonstrated mainly the 62 kDa protein in the same extracts. However, many protein bands were demonstrated in surface membrane (TSMP) and extracellular protein extracts with sera from only mildly infected animals. The protein patterns observed in all isolates of D. congolensis revealed global antigenic similarities and distinct differences

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

    Science.gov (United States)

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

    2010-06-01

    Puffer fish saxitoxin and tetrodotoxin binding protein (PSTBP) is a glycoprotein (200 kDa as a dimer) that we previously isolated from the plasma of Fugu pardalis (Yotsu-Yamashita et al., 2001). For the study on functions of PSTBP, here we examined distribution of homologous proteins to PSTBP in the plasma of seven species of puffer fish, and among the tissues of F. pardalis by Western blot analysis probed with a polyclonal IgG against unglycosylated PSTBP1 expressed in Echelichia coli. One or two major positive broad bands were detected at 105-140 kDa molecular weight range in the plasma (0.5 microg protein) of all species of puffer fish tested, while no band was detected in the plasma (5 microg protein) of fish other than puffer fish. Glycopeptidase F treated plasma of all species of puffer fish tested commonly showed the bands at approximately 42 kDa that was consistent to the molecular weight of unglycosylated PSTBP. These data suggest that puffer fish commonly possess glycoproteins homologous to PSTBP, but the sizes of N-glycan are specific to the species. Among soluble protein extracts (5 microg protein) from the tissues of F. pardalis, PSTBP was detected in all tissues examined, most prominently in heart, skin, and gall. Copyright 2009 Elsevier Ltd. All rights reserved.

  1. Comparison of T24H-his, GST-T24H and GST-Ts8B2 recombinant antigens in western blot, ELISA and multiplex bead-based assay for diagnosis of neurocysticercosis.

    Science.gov (United States)

    Hernández-González, Ana; Noh, John; Perteguer, María Jesús; Gárate, Teresa; Handali, Sukwan

    2017-05-15

    Currently, the reference standard assay for the serodiagnosis of neurocysticercosis (NCC) is the lentil lectin-bound glycoproteins/enzyme-linked immunoelectrotransfer blot (LLGP-EITB). The main disadvantage of this technique is the complexity of obtaining and purifying the LLGP extract. This could be solved by replacement with highly specific recombinant antigens from Taenia solium. Based on previous studies, we selected and produced the recombinant Ts8B2 and T24H proteins and applied them to three diagnostic techniques: western blot (WB), enzyme-linked immunosorbent assay (ELISA) and the multiplex bead-based assay (MBA). The Ts8B2 and T24H cDNA sequences were expressed in a prokaryotic system and the corresponding expression products purified; three recombinant proteins were further characterized: T24H-his, GST-T24H and GST-Ts8B2. The proteins on WB, ELISA and MBA were tested against 149 sera from patients with NCC confirmed by brain imaging, 40 sera from patients with other parasitic diseases, and 131 sera from US. individuals without evidence of neurocysticercosis (clinical/serological/brain imaging). The sensitivity and specificity of each antigen by WB were calculated by counting the number of true positive, false positive, true negative and false negative results. Using the receiver operating characteristic (ROC) curves, the cut-off values for the ELISA and MBA were established as well as the sensitivity and specificity of each assay. All three antigens showed a high sensitivity on WB in active NCC cases with two or more viable cysts and low sensitivity for cases with single viable cyst or calcified lesions and inactive NCC. WB showed the highest specificity and sensitivity out of the three diagnostic techniques. The recombinant T24H-his was the best diagnostic reagent in WB (100% sensitivity, 99.4% specificity), exhibiting similar results to the LLGP-EITB, against the same panel of NCC sera. The GST-T24H antigen worked better than the others in ELISA and MBA

  2. 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...... the gap to the more laborious nuclease protection experiments....

  3. Use of the REVERT® total protein stain as a loading control demonstrates significant benefits over the use of housekeeping proteins when analyzing brain homogenates by Western blot: An analysis of samples representing different gonadal hormone states.

    Science.gov (United States)

    Kirshner, Z Z; Gibbs, R B

    2018-01-30

    Western blot is routinely used to quantify differences in the levels of target proteins in tissues. Standard methods typically use measurements of housekeeping proteins to control for variations in loading and protein transfer. This is problematic, however, when housekeeping proteins also are affected by experimental conditions such as injury, disease, and/or gonadal hormone manipulations. Our goal was to evaluate an alternative and perhaps superior method for conducting Western blot analysis of brain tissue homogenates from rats with distinct physiologically relevant gonadal hormone states. Tissues were collected from the hippocampus, frontal cortex, and striatum of young adult female rats that either were ovariectomized to model surgical menopause, or were treated with the ovatotoxin 4-vinylcyclohexene diepoxide (VCD) to model transitional menopause. Tissues also were collected from rats with a normal estrous cycle killed at proestrus when estradiol levels are high, and at diestrus when estradiol levels are low. Western blot detection of α-tubulin, β-actin, and GAPDH was performed and were compared for sensitivity and reliability with a fluorescent total protein stain (REVERT®). Results show that the total protein stain was much less variable across samples and had a greater linear range than α-tubulin, β-actin, or GAPDH. The stain was stable and easy to use, and did not interfere with the immunodetection or multiplexed detection of the housekeeping proteins. In addition, we show that normalization of our data to total protein, but not to GAPDH, revealed significant differences in α-tubulin expression in the hippocampus as a function of treatment, and that gel-to-gel consistency in measuring differences between paired samples run on multiple gels was significantly better when data were normalized to total protein than when normalized to GAPDH. These results demonstrate that the REVERT® total protein stain can be used in Western blot analysis of brain

  4. 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...

  5. Detection of mu opioid receptor (MOPR) and its glycosylation in rat and mouse brains by western blot with anti-μC, an affinity-purified polyclonal anti-MOPR antibody.

    Science.gov (United States)

    Huang, Peng; Chen, Chongguang; Liu-Chen, Lee-Yuan

    2015-01-01

    Our experience demonstrates that it is difficult to identify MOPR in rat and mouse brains by western blot, in part due to low abundance of the receptor and a wide relative molecular mass (Mr) range of the receptor associated with its heterogeneous glycosylation states. Here, we describe generation and purification of anti-μC (a rabbit polyclonal anti-MOPR antibody), characterization of its specificity in immunoblotting of HA-tagged MOPR expressed in a cell line, and ultimately, unequivocal detection of the MOPR in brain tissues by western blot with multiple rigorous controls. In particular, using brain tissues from MOPR knockout (K/O) mice as the negative controls allowed unambiguous identification of the MOPR band, since the anti-MOPR antibody, even after affinity purification, recognizes nonspecific protein bands. The MOPR was resolved as a faint, broad, and diffuse band with a wide Mr range of 58-84 kDa depending on brain regions and species. Upon deglycosylation to remove N-linked glycans by PNGase F (but not Endo H), the MOPR became a dense and sharp band with Mr of ~43 kDa, close to the theoretical Mr of its deduced amino acid sequences. Thus, MOPRs in rodent brains are differentially glycosylated by complex type of N-linked glycans in brain region- and species-specific manners. Furthermore, we characterized the MOPR in an A112G/N38D-MOPR knockin mouse model that possesses the equivalent substitution of the A118G/N40D SNP in the human MOPR gene. The substitution removes one of the four and five N-linked consensus glycosylation sites of the mouse and human MOPR, respectively. We demonstrated that the Mr of the MOPR in A112G mouse brains was lower than that in wild-type mouse brains, and that the difference was due to lower degrees of N-linked glycosylation.

  6. 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.

  7. 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

  8. Ruthenium(II) complexes: DNA-binding, cytotoxicity, apoptosis, cellular localization, cell cycle arrest, reactive oxygen species, mitochondrial membrane potential and western blot analysis.

    Science.gov (United States)

    Li, Wei; Jiang, Guang-Bin; Yao, Jun-Hua; Wang, Xiu-Zhen; Wang, Ji; Han, Bing-Jie; Xie, Yang-Yin; Lin, Gan-Jian; Huang, Hong-Liang; Liu, Yun-Jun

    2014-11-01

    The aim of our study was to investigate DNA-binding and cytotoxic activity of the four new Ru(II) polypyridyl complexes [Ru(dmb)₂(HMHPIP)](ClO₄)₂ (1), [Ru(bpy)₂(HMHPIP)](ClO₄)₂ (2), [Ru(phen)₂(HMHPIP)](ClO₄)₂ (3) and [Ru(dmp)₂(HMHPIP)](ClO₄)₂ (4). The complexes interact with DNA through intercalative mode and show relatively high cytotoxic activity against A549 cells, no cytotoxicity toward MG-63 cells. Complexes 1-4 can enhance the levels of ROS in A549 cells and induce the decrease of the mitochondrial membrane potential. These complexes inhibit the cell growth in A549 cells at G0/G1 or S phase. Complex 3 activated caspase 7, and down-regulated the expression of the anti-apoptotic protein Bcl-2. Complexes 1-4 induce apoptosis in A549 cells through ROS-mediated mitochondrial dysfunction pathway. Crown Copyright © 2014. Published by Elsevier B.V. All rights reserved.

  9. Cost-effective and rapid lysis of Saccharomyces cerevisiae cells for quantitative western blot analysis of proteins, including phosphorylated eIF2α.

    Science.gov (United States)

    Lee, Su Jung; Ramesh, Rashmi; de Boor, Valerie; Gebler, Jan M; Silva, Richard C; Sattlegger, Evelyn

    2017-09-01

    The common method for liberating proteins from Saccharomyces cerevisiae cells involves mechanical cell disruption using glass beads and buffer containing inhibitors (protease, phosphatase and/or kinase inhibitors), followed by centrifugation to remove cell debris. This procedure requires the use of costly inhibitors and is laborious, in particular when many samples need to be processed. Also, enzymatic reactions can still occur during harvesting and cell breakage. As a result low-abundance and labile proteins may be degraded, and enzymes such as kinases and phosphatases may still modify proteins during and after cell lysis. We believe that our rapid sample preparation method helps overcome the above issues and offers the following advantages: (a) it is cost-effective, as no inhibitors and breaking buffer are needed; (b) cell breakage is fast (about 15 min) since it only involves a few steps; (c) the use of formaldehyde inactivates endogenous proteases prior to cell lysis, dramatically reducing the risk of protein degradation; (d) centrifugation steps only occur prior to cell lysis, circumventing the problem of losing protein complexes, in particular if cells were treated with formaldehyde intended to stabilize and capture large protein complexes; and (e) since formaldehyde has the potential to instantly terminate protein activity, this method also allows the study of enzymes in live cells, i.e. in their true physiological environment, such as the short-term effect of a drug on enzyme activity. Taken together, the rapid sample preparation procedure provides a more accurate snapshot of the cell's protein content at the time of harvesting. Copyright © 2017 John Wiley & Sons, Ltd. Copyright © 2017 John Wiley & Sons, Ltd.

  10. Indeterminate HIV western blot test result.

    Science.gov (United States)

    Barnett, Deborah L; Duffus, Wayne A

    2008-12-01

    An IWB is a test result that creates uncertainty. Explaining this result to patients can cause provider discomfort but the more the provider practices, the comfort level increases. HIV diagnosis is no longer considered a death sentence. It has become a manageable disease when patients enter care early. That is why it is important to encourage screening for all. The earlier a patient is linked to care the better the health outcome and this prevents ongoing HIV transmission.

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

    Science.gov (United States)

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

    2013-12-01

    An accurate and rapid serologic method to differentiate HIV-2 from HIV-1 infection is required since the confirmatory HIV-1 Western Blot (WB) may demonstrate cross-reactivity with HIV-2 antibodies. To evaluate the performance of the Bio-Rad Multispot HIV-1/HIV-2 rapid assay as a supplemental test to correctly identify HIV-2 infection and identify HIV-1 WB cross-reactivity with HIV-2 in clinical samples tested at an academic medical center. Between August 2008 and July 2012, clinical samples were screened for HIV using either 3rd- or 4th-generation HIV-1/2 antibody or combination antibody and HIV-1 p24 antigen assays, respectively. All repeatedly reactive samples were reflexed for Multispot rapid testing. Multispot HIV-2 and HIV-1 and HIV-2-reactive samples were further tested using an HIV-2 immunoblot assay and HIV-1 or HIV-2 RNA assays when possible. The HIV-1 WB was performed routinely for additional confirmation and to assess for HIV-2 antibody cross-reactivity. Of 46,061 samples screened, 890 (89.6%) of 993 repeatedly reactive samples were also Multispot-reactive: 882 for HIV-1; three for only HIV-2; and five for both HIV-1 and HIV-2. All three HIV-2-only Multispot-positives along with a single dually reactive HIV-1/2 Multispot-positive were also HIV-2 immunoblot-positive; the latter was HIV-1 RNA negative and HIV-2 RNA positive. The Multispot rapid test performed well as a supplemental test for HIV-1/2 diagnostic testing. Four new HIV-2 infections (0.45%) were identified from among 890 Multispot-reactive tests. The use of HIV-1 WB alone to confirm HIV-1/2 screening assays may underestimate the true prevalence of HIV-2 infection in the United States. Copyright © 2013 Elsevier B.V. All rights reserved.

  12. Evaluation of supplemental testing with the Multispot HIV-1/HIV-2 Rapid Test and APTIMA HIV-1 RNA Qualitative Assay to resolve specimens with indeterminate or negative HIV-1 Western blots.

    Science.gov (United States)

    Linley, Laurie; Ethridge, Steven F; Oraka, Emeka; Owen, S Michele; Wesolowski, Laura G; Wroblewski, Kelly; Landgraf, Kenneth M; Parker, Monica M; Brinson, Myra; Branson, Bernard M

    2013-12-01

    The use of Western blot (WB) as a supplemental test after reactive sensitive initial assays can lead to inconclusive or misclassified HIV test results, delaying diagnosis. To determine the proportion of specimens reactive by immunoassay (IA) but indeterminate or negative by WB that could be resolved by alternative supplemental tests recommended under a new HIV diagnostic testing algorithm. Remnant HIV diagnostic specimens that were reactive on 3rd generation HIV-1/2 IA and either negative or indeterminate by HIV-1 WB from 11 health departments were tested with the Bio-Rad Multispot HIV-1/HIV-2 Rapid Test (Multispot) and the Gen-Probe APTIMA HIV-1 RNA Qualitative Assay (APTIMA). According to the new testing algorithm, 512 (89.8%) specimens were HIV-negative, 55 (9.6%) were HIV-1 positive (including 19 [3.3%] that were acute HIV-1 and 9 [1.6%] that were positive for HIV-1 by Multispot but APTIMA-negative), 2 (0.4%) were HIV-2 positive, and 1 (0.2%) was HIV-positive, type undifferentiated. 47 (21.4%) of the 220 WB-indeterminate and 8 (2.3%) of the 350 WB-negative specimens were HIV-1 positive. Applying the new HIV diagnostic algorithm retrospectively to WB-negative and indeterminate specimens, the HIV infection status could be established for nearly all of the specimens. IA-reactive HIV-infected persons with WB-negative results had been previously misclassified as uninfected, and HIV diagnosis was delayed for those with WB-indeterminate specimens. These findings underscore the limitations of the WB to confirm HIV infection after reactive results from contemporary 3rd or 4th generation IAs that can detect HIV antibodies several weeks sooner than the WB. Published by Elsevier B.V.

  13. A western boundary current eddy characterisation study

    Science.gov (United States)

    Ribbe, Joachim; Brieva, Daniel

    2016-12-01

    The analysis of an eddy census for the East Australian Current (EAC) region yielded a total of 497 individual short-lived (7-28 days) cyclonic and anticyclonic eddies for the period 1993 to 2015. This was an average of about 23 eddies per year. 41% of the tracked individual cyclonic and anticyclonic eddies were detected off southeast Queensland between about 25 °S and 29 °S. This is the region where the flow of the EAC intensifies forming a swift western boundary current that impinges near Fraser Island on the continental shelf. This zone was also identified as having a maximum in detected short-lived cyclonic eddies. A total of 94 (43%) individual cyclonic eddies or about 4-5 per year were tracked in this region. The census found that these potentially displaced entrained water by about 115 km with an average displacement speed of about 4 km per day. Cyclonic eddies were likely to contribute to establishing an on-shelf longshore northerly flow forming the western branch of the Fraser Island Gyre and possibly presented an important cross-shelf transport process in the life cycle of temperate fish species of the EAC domain. In-situ observations near western boundary currents previously documented the entrainment, off-shelf transport and export of near shore water, nutrients, sediments, fish larvae and the renewal of inner shelf water due to short-lived eddies. This study found that these cyclonic eddies potentially play an important off-shelf transport process off the central east Australian coast.

  14. An ART field study in western Nepal.

    Science.gov (United States)

    Yee, R

    2001-04-01

    To evaluate the effectiveness of three different types of glass ionomer restorations placed in children's teeth using the ART technique two years after placement by either dentists or primary health care workers. The rural district of Palpa in western Nepal. An opportunistic non-randomised field study, commenced in April 1997. Three different glass ionomer restorative cements were used: Fuji IX (GC), Dentsply Baseline (DENTSPLY/DeTrey), and S.S. WhiteR (S.S. White). The restorations were assessed two years later by an independent evaluator, who was blind to the type of restorations used and also blind to the operators. Four primary health care workers and two dentists restored 163 primary and permanent teeth using hand instruments. One and two surface restorations were placed in 105 students ranging from 3 to 19 years of age from seven schools. After two years, 79 (75%) of the restorations survived and 27 (25%) failed, while 100% of 21 evaluated single surface restorations placed on permanent molars with Fuji IX survived. There was no statistically significant difference in the survival of the restorations placed by dentists and placed by primary health care workers, which corroborates that both groups are both capable of performing the technique satisfactorily. ART is very effective for one surface permanent molars using improved glass ionomer cements designed for the ART technique.

  15. Glycosaminoglycan blotting and detection after electrophoresis separation.

    Science.gov (United States)

    Volpi, Nicola; Maccari, Francesca

    2015-01-01

    Separation of glycosaminoglycans (GAGs) by electrophoresis and their characterization to the microgram level are integral parts of biochemical research. Their blotting on membranes after electrophoresis offers the advantage to perform further analysis on single separated species such as identification with antibodies and/or recovery of single band. A method for the blotting and immobilizing of several nonsulfated and sulfated complex GAGs on membranes made hydrophilic and positively charged by cationic detergent after their separation by conventional agarose-gel electrophoresis is illustrated. This approach to the study of these complex macromolecules utilizes the capacity of agarose-gel electrophoresis to separate single species of polysaccharides from mixtures and the membrane technology for further preparative and analytical uses. Nitrocellulose membranes are derivatized with the cationic detergent cetylpyridinium chloride (CPC) and mixtures of GAGs are capillary blotted after their separation in agarose-gel electrophoresis. Single purified species of variously sulfated polysaccharides are transferred on derivatized membranes with an efficiency of 100 % and stained with alcian blue (irreversible staining) and toluidine blue (reversible staining). This enables a lower amount limit of detection of 0.1 μg. Nonsulfated polyanions, for example hyaluronic acid (HA), may also be transferred to membranes with a limit of detection of approximately 0.1-0.5 μg after irreversible or reversible staining. The membranes may be stained with reversible staining and the same lanes used for immunological detection or other applications.

  16. 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…

  17. Studies in western yellow pine nursery practice

    Science.gov (United States)

    Donald R. Brewster; J. A. Larsen

    1925-01-01

    In 1912 and 1913, when nursery experiments were started under direction of the then "Priest River'' Forest Experiment Station, at Priest River, Idaho, and elsewhere, western yellow pine (Pinus ponderosa) was one of the principal species being planted on a large scale in the northern Rocky Mountain region and millions of plants were being raised each year...

  18. Western Wind and Solar Integration Study Phase 3: Technical Overview

    Energy Technology Data Exchange (ETDEWEB)

    2015-11-01

    Technical fact sheet outlining the key findings of Phase 3 of the Western Wind and Solar Integration Study (WWSIS-3). NREL and GE find that with good system planning, sound engineering practices, and commercially available technologies, the Western grid can maintain reliability and stability during the crucial first minute after grid disturbances with high penetrations of wind and solar power.

  19. Evaluation of a genus-specific ELISA and a commercial Aspergillus Western blot IgG® immunoblot kit for the diagnosis of aspergillosis in common bottlenose dolphins (Tursiops truncatus).

    Science.gov (United States)

    Desoubeaux, Guillaume; Le-Bert, Carolina; Fravel, Vanessa; Clauss, Tonya; Delaune, Alexa J; Soto, Jeny; Jensen, Eric D; Flower, Jennifer E; Wells, Randall; Bossart, Gregory D; Cray, Carolyn

    2017-12-08

    Aspergillosis is a fungal infection with high mortality and morbidity rates. As in humans, its definitive diagnosis is difficult in animals, and thus new laboratory tools are required to overcome the diagnostic limitations due to low specificity and lack of standardization. In this study of common bottlenose dolphins (Tursiops truncatus), we evaluated the diagnostic performance of a new commercial immunoblot kit that had been initially developed for the serologic diagnosis of chronic aspergillosis in humans. Using this in a quantitative approach, we first established its positive cutoff within an observation cohort of 32 serum samples from dolphins with "proven" or "probable" diagnosis of aspergillosis and 55 negative controls. A novel enzyme-linked immunosorbent assay (ELISA) test was also developed for detecting anti-Aspergillus antibodies, and results were compared between the two assays. Overall, the diagnostic performance of immunoblot and ELISA were strongly correlated (P < .0001). The former showed lower sensitivity (65.6% versus 90.6%), but higher specificity (92.7% vs. 69.1%), with no cross-reaction with other fungal infections caused by miscellaneous non-Aspergillus genera. When assessing their use in a validation cohort, the immunoblot kit and the ELISA enabled positive diagnosis before mycological cultures in 42.9% and 33.3% subjects addressed for suspicion of aspergillosis, respectively. There was also significant impact of antifungal treatment on the results of the two tests (P < .05). In all, these new serological methods show promise in aiding in the diagnosis of aspergillosis in dolphins, and illustrate the opportunity to adapt commercial reagents directed for human diagnostics to detect similar changes in other animals. © The Author 2017. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  20. Phenylglyoxal-based visualization of citrullinated proteins on Western blots

    NARCIS (Netherlands)

    Hensen, S.M.; Boelens, W.C.; Bonger, K.M.; Cruchten, R.T.P. van; Delft, F.L. van; Pruijn, G.J.

    2015-01-01

    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

  1. 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.

  2. Western Wind and Solar Integration Study: Phase 2 (Presentation)

    Energy Technology Data Exchange (ETDEWEB)

    Lew, D.; Brinkman, G.; Ibanez, E.; Lefton, S.; Kumar, N.; Venkataraman, S.; Jordan, G.

    2013-09-01

    This presentation summarizes the scope and results of the Western Wind and Solar Integration Study Phase 2, which examined operational impacts of high penetrations of variable renewable generation in the West.

  3. Perception of Western Musical Modes: A Chinese Study

    Directory of Open Access Journals (Sweden)

    Lele Fang

    2017-10-01

    Full Text Available The major mode conveys positive emotion, whereas the minor mode conveys negative emotion. However, previous studies have primarily focused on the emotions induced by Western music in Western participants. The influence of the musical mode (major or minor on Chinese individuals’ perception of Western music is unclear. In the present experiments, we investigated the effects of musical mode and harmonic complexity on psychological perception among Chinese participants. In Experiment 1, the participants (N = 30 evaluated 24 musical excerpts in five dimensions (pleasure, arousal, dominance, emotional tension, and liking. In Experiment 2, the participants (N = 40 evaluated 48 musical excerpts. Perceptions of the musical excerpts differed significantly according to mode, even if the stimuli were Western musical excerpts. The major-mode music induced greater pleasure and arousal and produced higher liking ratings than the minor-mode music, whereas the minor-mode music induced greater tension than the major-mode music. Mode did not influence the dominance rating. Perception of Western music was not influenced by harmonic complexity. Moreover, preference for musical mode was influenced by previous exposure to Western music. These results confirm the cross-cultural emotion induction effects of musical modes in Western music.

  4. Perception of Western Musical Modes: A Chinese Study.

    Science.gov (United States)

    Fang, Lele; Shang, Junchen; Chen, Nan

    2017-01-01

    The major mode conveys positive emotion, whereas the minor mode conveys negative emotion. However, previous studies have primarily focused on the emotions induced by Western music in Western participants. The influence of the musical mode (major or minor) on Chinese individuals' perception of Western music is unclear. In the present experiments, we investigated the effects of musical mode and harmonic complexity on psychological perception among Chinese participants. In Experiment 1, the participants (N = 30) evaluated 24 musical excerpts in five dimensions (pleasure, arousal, dominance, emotional tension, and liking). In Experiment 2, the participants (N = 40) evaluated 48 musical excerpts. Perceptions of the musical excerpts differed significantly according to mode, even if the stimuli were Western musical excerpts. The major-mode music induced greater pleasure and arousal and produced higher liking ratings than the minor-mode music, whereas the minor-mode music induced greater tension than the major-mode music. Mode did not influence the dominance rating. Perception of Western music was not influenced by harmonic complexity. Moreover, preference for musical mode was influenced by previous exposure to Western music. These results confirm the cross-cultural emotion induction effects of musical modes in Western music.

  5. Study physico-chemical of the sand of the western ERG (Western South Algeria)

    Energy Technology Data Exchange (ETDEWEB)

    Allam, M.; Tafraoui, A. [Faculty of sciences and technology, University of Bechar (Algeria)], email: allammessaouda@yahoo.fr

    2011-07-01

    Silica is gaining increasing importance as it is the base for the production of pure silicon, for which several applications are under development in the electronic and solar energy sectors. The aim of this study is to characterize the sand taken from the Western Erg of Algeria to determine the percentage of silicon it contains. Characterization was done through physical analysis to determine the granulometry of the sand. A chemical analysis was next performed, using diffraction of X-rays and a scanning electron microscope to determine the chemical composition of the sand. Results showed that the sand is mainly made of quartz in the form of rounded and subbarrondis grains and that silicon is prevalent, accounting for 98% of the composition. This study demonstrated that sand from the Western Erg of Algeria is rich in silicon and could be used for silicon production.

  6. Western gulf culture-density study-early results

    Science.gov (United States)

    Mohd S. Rahman; Michael G. Messina; Richard F. Fisher; Alan B. Wilson; Nick Chappell; Conner Fristoe; Larry Anderson

    2006-01-01

    The Western Gulf Culture-Density Study is a collaborative research effort between Texas A&M University and five forest products companies to examine the effects of early silvicultural treatment intensity and a wide range of both densities and soil types on performance of loblolly pine. The study tests 2 silvicultural intensities, 5 planting densities (200 to 1,200...

  7. Western Wind and Solar Integration Study: Hydropower Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Acker, T.; Pete, C.

    2012-03-01

    The U.S. Department of Energy's (DOE) study of 20% Wind Energy by 2030 was conducted to consider the benefits, challenges, and costs associated with sourcing 20% of U.S. energy consumption from wind power by 2030. This study found that with proactive measures, no insurmountable barriers were identified to meet the 20% goal. Following this study, DOE and the National Renewable Energy Laboratory (NREL) conducted two more studies: the Eastern Wind Integration and Transmission Study (EWITS) covering the eastern portion of the U.S., and the Western Wind and Solar Integration Study (WWSIS) covering the western portion of the United States. The WWSIS was conducted by NREL and research partner General Electric (GE) in order to provide insight into the costs, technical or physical barriers, and operational impacts caused by the variability and uncertainty of wind, photovoltaic, and concentrated solar power when employed to serve up to 35% of the load energy in the WestConnect region (Arizona, Colorado, Nevada, New Mexico, and Wyoming). WestConnect is composed of several utility companies working collaboratively to assess stakeholder and market needs to and develop cost-effective improvements to the western wholesale electricity market. Participants include the Arizona Public Service, El Paso Electric Company, NV Energy, Public Service of New Mexico, Salt River Project, Tri-State Generation and Transmission Cooperative, Tucson Electric Power, Xcel Energy and the Western Area Power Administration.

  8. Slash smoke dispersal over western Oregon...a case study

    Science.gov (United States)

    John D. Dell; Franklin R. Ward; Robert E. Lynott

    1970-01-01

    Smoke from slash burns in the Cascade Mountains during a 3-day period ofstable air conditions at lower elevations in October 1969 added little to existing air pollution in the Willamette Valley, in western Oregon. Aerial observations and weather data analysis determined that slash smoke dispersed eastward — away from the Valley. Studies of this type can help improve...

  9. Use of social media by Western European hospitals: longitudinal study.

    Science.gov (United States)

    Van de Belt, Tom H; Berben, Sivera A A; Samsom, Melvin; Engelen, Lucien J L P G; Schoonhoven, Lisette

    2012-05-01

    Patients increasingly use social media to communicate. Their stories could support quality improvements in participatory health care and could support patient-centered care. Active use of social media by health care institutions could also speed up communication and information provision to patients and their families, thus increasing quality even more. Hospitals seem to be becoming aware of the benefits social media could offer. Data from the United States show that hospitals increasingly use social media, but it is unknown whether and how Western European hospitals use social media. To identify to what extent Western European hospitals use social media. In this longitudinal study, we explored the use of social media by hospitals in 12 Western European countries through an Internet search. We collected data for each country during the following three time periods: April to August 2009, August to December 2010, and April to July 2011. We included 873 hospitals from 12 Western European countries, of which 732 were general hospitals and 141 were university hospitals. The number of included hospitals per country ranged from 6 in Luxembourg to 347 in Germany. We found hospitals using social media in all countries. The use of social media increased significantly over time, especially for YouTube (n = 19, 2% to n = 172, 19.7%), LinkedIn (n =179, 20.5% to n = 278, 31.8%), and Facebook (n = 85, 10% to n = 585, 67.0%). Differences in social media usage between the included countries were significant. Social media awareness in Western European hospitals is growing, as well as its use. Social media usage differs significantly between countries. Except for the Netherlands and the United Kingdom, the group of hospitals that is using social media remains small. Usage of LinkedIn for recruitment shows the awareness of the potential of social media. Future research is needed to investigate how social media lead to improved health care.

  10. 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.

  11. Retina-derived fetuin (RDF): analysis by immunocytochemistry, reverse transcriptase-polymerase chain reaction and Southern blot.

    Science.gov (United States)

    Sheedlo, H J; Krishnamoorthy, R S; Nelson, T S; Agarwal, N S; Liu, J S; Roque, R S; Wordinger, R J; Jaynes, C D; Brun-Zinkelnagel, A M; O'Brian, P; Aschenbrenner, J E; Turner, J E

    1999-12-01

    This study was undertaken to determine the presence of retina-derived fetuin (RDF) protein and its message in retinal tissues and retinal pigment epithelial (RPE) cells. The techniques utilized in this study included light micros-copy, immunochemistry, Western blot, reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blot. Retinal tissues and sections from embryonic, early postnatal and adult normal rats and retinal pigment epithe-lial (RPE) cells from postnatal rats were immunostained for fetuin with a polyclonal fetuin antibody and a peroxidase conjugated-secondary antibody using immunocytochemistry and Western blot analyses. The cDNA generated from RNA isolated from early postnatal rat retinas and RPE was probed with primers for rat fetuin, amplified by PCR and the PCR products were analyzed by Southern blot. Fetuin (RDF) was immunolocalized to cells of the neuroepithelium in retinas of early postnatal rats and most prominently in the nuclei and perinuclear region of cultured neonatal rat RPE cells. In adult retinas, ganglion cells, inner segments of photoreceptor cells, some components of the outer plexiform layer, ganglion cells and optic nerve processes were immunoreactive for the fetuin protein. As shown by Western blot, fetuin (RDF) was higher in embryonic and early postnatal retinas than in late postnatal retinas, indicating that this protein may be developmentally regulated. Using RT-PCR, the message for rat fetuin was demonstrated in the retina and RPE of normal postnatal rats. Southern blot confirmed that the PCR product from the retina and RPE was generated from rat fetuin mRNA as well as from rat liver, the primary source of fetuin. Fetuin, termed retina-derived fetuin (RDF), is reported for the first time in retinal tissues. Fetuin is a cysteine protease inhibitor that may play a role in support of neuronal cell survival during early retinal development and the maintenance of neuronal activity. RDF may interact with other growth

  12. Diyabetik\tSıçanların\tTestis\tDokusunda\tVEGF\tve\tBcl-2\tEkspresyon\tDüzeylerinin İmmünohistokimya\tve\tWestern\tBlot\tYöntemleri\tİle\tİncelenmesi

    Directory of Open Access Journals (Sweden)

    Sevgi İrtegün

    2016-12-01

    Full Text Available Amaç: Bu\tçalışmada\tdiyabetik\tsıçanların tetis\tdokusunda\tanjiogenezi\tve\tvasküler\tgeçirgenliği\tstimule\teden\tvasküler\tendotel\tbüyüme faktörünün (VEGF ve apoptozis regülasyonunda önemli bir rol oynayan B-cell lymphoma-2’ nin (Bcl-2 ekspresyon düzeylerini araştırmayı\tamaçladık. Yöntemler: Bu\tçalışmada\t20\tadet\terişkin\tWistar\talbino\tsıçan\tkullanıldı.\tSıçanlar\tkontrol\tve\tDiabetes\tmellitus\t(DM\tolmak\tüzere\t2\teşit gruba\tayrıldı.\tDM\toluşturmak\tiçin\tsitrat\ttamponunda\tçözülmüş\ttek\tdoz\tStreptozotosin\t(STZ\t(55mg/kg\tintraperitoneal\tolarak\tverildi. %10’\tluk\tformaldehit\tsolüsyonuna\tatılan\ttestis\tdokuları\trutin\tparafin\ttakiplerinden\tsonra\thistopatolojik\tolarak\tincelendi.\tVEGF\tve\tBcl-2 protein\tdüzeyleri\timmünohistokimya\tve\tWestern\tBlot\työntemleriyle\tölçüldü. Bulgular: Diyabetik\tgruba\tait\tspermatik\thücrelerin\tbazılarında\tdejenerasyon,\tçekirdekte\tküçülme\tve\tyer\tyer\tpiknosis\tgözlendi.\tDiyabet sonucu Sertoli hücrelerinde yapısal değişiklikler saptanırken, tubuller arasındaki damarlarda dilatasyon ve hemoraji gözlendi. Diyabetin\tetkisi\tile\ttestis\tdokusunda\tVEGF\tekspresyonunun\tbloke\tolduğu,\tBcl-2\tekspresyonunun\tise\tazaldığı\tsaptandı. Sonuç: Diyabetik testis dokularında VEGF ekspresyonunun bloke olmasının yetersiz anjiogenez ve vasküler permeabiliteye neden olabileceği olasıdır. Ayrıca diyabetik testis dokularında anti-apoptotik protein olan Bcl-2 ekspresyonundaki azalmanın diyabetin testiküler dokuda\tapotozis\tregülasyonunun\tbozulmasına\tneden\tolabileceği\tdüşünülmektedir.

  13. Geography Teacher Candidates' Experiences of Field Study in Western Anatolia: A Qualitative Study

    Science.gov (United States)

    Balci, Ali

    2012-01-01

    The aim of this study is to establish expectations of 5th grade students from Marmara University's Department of Geography Teaching on geographical field study in Western Anatolia. For this reason, a field study trip was organized to Western Anatolia. A survey, which was consisted of open-ended questions, was prepared by experts and it was…

  14. 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

  15. Western Wind and Solar Integration Study Phase 2: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Lew, D.; Brinkman, G.; Ibanez, E.; Hodge, B.-M.; King, J.

    2012-09-01

    The Western Wind and Solar Integration Study (WWSIS) investigates the impacts of high penetrations of wind and solar power into the Western Interconnection of the United States. WWSIS2 builds on the Phase 1 study but with far greater refinement in the level of data inputs and production simulation. It considers the differences between wind and solar power on systems operations. It considers mitigation options to accommodate wind and solar when full costs of wear-and-tear and full impacts of emissions rates are taken into account. It determines wear-and-tear costs and emissions impacts. New data sets were created for WWSIS2, and WWSIS1 data sets were refined to improve realism of plant output and forecasts. Four scenarios were defined for WWSIS2 that examine the differences between wind and solar and penetration level. Transmission was built out to bring resources to load. Statistical analysis was conducted to investigate wind and solar impacts at timescales ranging from seasonal down to 5 minutes.

  16. Western Wind and Solar Integration Study Phase 2 (Presentation)

    Energy Technology Data Exchange (ETDEWEB)

    Lew, D.; Brinkman, G.; Ibanez, E.; Kumar, N.; Lefton, S.; Jordan, G.; Venkataraman, S.; King, J.

    2013-06-01

    This presentation accompanies Phase 2 of the Western Wind and Solar Integration Study, a follow-on to Phase 1, which examined the operational impacts of high penetrations of variable renewable generation on the electric power system in the West and was one of the largest variable generation studies to date. High penetrations of variable generation can induce cycling of fossil-fueled generators. Cycling leads to wear-and-tear costs and changes in emissions. Phase 2 calculated these costs and emissions, and simulated grid operations for a year to investigate the detailed impact of variable generation on the fossil-fueled fleet. The presentation highlights the scope of the study and results.

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

    Science.gov (United States)

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

    2011-02-01

    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.

  18. The Western Wind and Solar Integration Study Phase 2

    Energy Technology Data Exchange (ETDEWEB)

    Lew, Debra [National Renewable Energy Lab. (NREL), Golden, CO (United States); Brinkman, Greg [National Renewable Energy Lab. (NREL), Golden, CO (United States); Ibanez, E. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Florita, A. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Heaney, M. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Hodge, B. -M. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Hummon, M. [National Renewable Energy Lab. (NREL), Golden, CO (United States); Stark, G. [National Renewable Energy Lab. (NREL), Golden, CO (United States); King, J. [RePPAE; Lefton, S. A. [Intertek-APTECH, Houston, TX (United States); Kumar, N. [Intertek-APTECH, Houston, TX (United States); Agan, D. [Intertek-APTECH, Houston, TX (United States); Jordan, G. [GE Energy, Fairfield, CT (United States); Venkataraman, S. [GE Energy, Fairfield, CT (United States)

    2013-09-01

    The electric grid is a highly complex, interconnected machine, and changing one part of the grid can have consequences elsewhere. Adding wind and solar affects the operation of the other power plants and adding high penetrations can induce cycling of fossil-fueled generators. Cycling leads to wear-and-tear costs and changes in emissions. Phase 2 of the Western Wind and Solar Integration Study (WWSIS-2) evaluated these costs and emissions and simulated grid operations for a year to investigate the detailed impact of wind and solar on the fossil-fueled fleet. This built on Phase 1, one of the largest wind and solar integration studies ever conducted, which examined operational impacts of high wind and solar penetrations in the West(GE Energy 2010).

  19. The Western Wind and Solar Integration Study Phase 2

    Energy Technology Data Exchange (ETDEWEB)

    Lew, D.; Brinkman, G.; Ibanez, E.; Hodge, B. M.; Hummon, M.; Florita, A.; Heaney, M.

    2013-09-01

    The electric grid is a highly complex, interconnected machine, and changing one part of the grid can have consequences elsewhere. Adding wind and solar affects the operation of the other power plants and adding high penetrations can induce cycling of fossil-fueled generators. Cycling leads to wear-and-tear costs and changes in emissions. Phase 2 of the Western Wind and Solar Integration Study (WWSIS-2) evaluated these costs and emissions and simulated grid operations for a year to investigate the detailed impact of wind and solar on the fossil-fueled fleet. This built on Phase 1, one of the largest wind and solar integration studies ever conducted, which examined operational impacts of high wind and solar penetrations in the West.

  20. A contribution to mayfly studies of Western Mongolia (Insecta, Ephemeroptera).

    Science.gov (United States)

    Erdenee, Bolortsetseg; Maasri, Alain; Gelhaus, Jon K; Bayartogtokh, Badamdorj

    2016-01-01

    Streams in the Mongolian Altai Mountains are mostly fed from glaciers and are extreme conditions for mayflies because of high elevation, low temperatures and low annual precipitation. Previous information about mayflies of Western Mongolia is scarce, but with this study a total of 38 species belonging to 26 genera and subgenera and 8 families of mayflies has been recorded in the Mongolian Altai region. Study material was entirely imagos and collected from 78 sites during expeditions led by the Mongolian Aquatic Insect Survey in 2008, 2009 and 2010. Raptobaetopus tenellus, Caenis luctuosa and Caenis rivulorum are recorded as new to the fauna of Mongolia, and there are new distribution records for Ameletus montanus, Baetis (Acentrella) lapponica, Baetis sibiricus, Baetis (Labiobaetis) attrebatinus, Centroptilum luteolum, Procloeon pennulatum, Ephemerella aurivillii, Serratella setigera, Ephemera sachalinensis, Ecdyonurus (Afronurus) abracadabrus, Cinygmula kurenzovi, Ecdyonurus (Afghanurus) vicinus and Epeorus (Belovius) pellucidus from the Mongolian Altai region. Baetis vernus and Ephemerella aurivillii are the most frequently encountered species in this region.

  1. The Applicability of Western Socio-Legal Frameworks to the Study of Negotiation in Chinese Society

    Science.gov (United States)

    Chung, Mona; Ingleby, Richard

    2011-01-01

    Theoretical frameworks for the examination of negotiation generated by Western academics do not easily translate to Chinese society because of fundamental differences between Western and Chinese society. Attempts to study negotiation in Chinese society and to improve negotiation between Chinese and Western business people are themselves…

  2. Failure analysis of blots for diesel engine intercooler

    Science.gov (United States)

    Ren, Ping; Li, Zongquan; Wu, Jiangfei; Guo, Yibin; Li, Wanyou

    2017-05-01

    In diesel generating sets, it will lead to the abominable working condition if the fault couldn’t be recovered when the bolt of intercooler cracks. This paper aims at the fault of the blots of diesel generator intercooler and completes the analysis of the static strength and fatigue strength. Static intensity is checked considering blot preload and thermal stress. In order to obtain the thermal stress of the blot, thermodynamic of intercooler is calculated according to the measured temperature. Based on the measured vibration response and the finite element model, using dynamic load identification technique, equivalent excitation force of unit was solved. In order to obtain the force of bolt, the excitation force is loaded into the finite element model. By considering the thermal stress and preload as the average stress while the mechanical stress as the wave stress, fatigue strength analysis has been accomplished. Procedure of diagnosis is proposed in this paper. Finally, according to the result of intensity verification the fatigue failure is validation. Thereby, further studies are necessary to verification the result of the intensity analysis and put forward some improvement suggestion.

  3. Archaeomagnetic Study performed on Early Medieval Buildings from western France

    Science.gov (United States)

    Chauvin, A.; Lanos, P.; Dufresne, P.; Blain, S.; Guibert, P.; Oberlin, C.; Sapin, C.

    2009-05-01

    A multiple dating study, involving a collaboration between specialists of dating techniques (thermoluminescence (TL) and radiocarbon), historians of art and archaeologists, has been carried out on several early medieval buildings from western France. The early medieval period is not well known especially in France where there is a lack of visible evidence that identifies pre-Romanesque architecture. The majority of buildings to have survived from this period are religious ones, considered important enough to be made of strong, non-perishable material such as stone or brick, as for example the churches of Notre-Dame-sous- Terre in the Mont-Saint-Michel or St Martin in Angers. Due to their significance in architectural history, it is imperative to position them accurately in the chronology of the history of art. Bricks are often used to build up round-headed arches or to reinforce the frame of a wall with bonding courses in those churches. TL dating and archeomagnetic analysis were performed on cores drilled within bricks while radiocarbon dating were undertaken on coals found within mortars. In order to increase the number of data during the early Middle Ages, archeointensity determinations using the classical Thellier technique with anisotropy of thermal remanence and cooling rate corrections were performed. Archaeomagnetic directions were used to recognize the firing position of bricsk during manufacture. Reliable and precise ages were obtained on the church Notre-Dame-sous-Terre; they indicate two phases of building in 950±50AD and 990±50AD. Mean archeointensities obtained on 17 (21) samples from the first (second) phases appears very closed 69.1±1.2 and 68.3±1.6 microTesla. Ages and archeomagnetic results obtained on 4 other sites will be presented and compared to the available data in western Europe.

  4. Zinc Blotting Assay for Detection of Zinc-Binding Prolamin in Barley (Hordeum vulgare) Grain

    DEFF Research Database (Denmark)

    Uddin, Mohammad Nasir; Langkilde, Ane; Vincze, Éva

    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-bindin...... formation. The dithizone staining method gave similar reproducibility to the radioactive blotting. The detected zinc-binding protein was identified as B-hordein by Western blotting......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...

  5. Epidemiological study of road traffic accident cases from Western Nepal

    Directory of Open Access Journals (Sweden)

    Mishra Badrinarayan

    2010-01-01

    Full Text Available Background: Road Traffic Accident (RTA is one among the top five causes of morbidity and mortality in South-East Asian countries. [1] Its socioeconomic repercussions are a matter of great concern. Efficient addressing of the issue requires quality information on different causative factors. Research Question: What are different epidemiological determinants of RTA in western Nepal? Objective: To examine the factors associated with RTA. Study Design: Prospective observational. Setting: Study was performed in a tertiary healthcare delivery institute in western Nepal. Participants: 360 victims of RTA who reported to Manipal Teaching hospital in one year. Study Variables: Demographic, human, vehicular, environmental and time factors. Statistical analysis: Percentages, linear and logarithmic trend and Chi-square. Results: Most of the victims i.e. 147 (40.83% were young (15 to 30 years; from low i.e. 114 (31.66% and mid i.e. 198 (55% income families and were passengers i.e. 153 (42.50% and pedestrians i.e. 105 (29.16%. Sever accidents leading to fatal outcome were associated with personal problems (P< 0.01, χ2 - 8.03, recent or on-day conflicts (P< 0.001, χ2 - 18.88 and some evidence of alcohol consumptions (P< 0.001, χ2 - 30.25. Increased prevalence of RTA was also noticed at beginning i.e. 198 (55% and end i.e. 69 (19.16% of journey; in rainy and cloudy conditions (269 i.e. 74.72% and in evening hours (3 to 7 p.m. 159 i.e. 44.16%. Out of 246 vehicles involved; 162 ( 65.85% were old and ill maintained. The contributions of old vehicle to fatal injuries were 33 (50%. Head injury was found in 156 (43.33 % cases and its associated case fatality rate was 90.90%. In spite of a good percentage receiving first aid i.e. 213 (59.16% after RTA; there was a notable delay (174 i.e. 48.33% admitted after 6 h in shifting the cases to the hospitals. The estimated total days lost due to hospital stay was 4620 with an average of 12.83 days per each case. Conclusion

  6. A contribution to mayfly studies of Western Mongolia (Insecta, Ephemeroptera

    Directory of Open Access Journals (Sweden)

    Bolortsetseg Erdenee

    2016-12-01

    Full Text Available Streams in the Mongolian Altai Mountains are mostly fed from glaciers and are extreme conditions for mayflies because of high elevation, low temperatures and low annual precipitation. Previous information about mayflies of Western Mongolia is scarce, but with this study a total of 38 species belonging to 26 genera and subgenera and 8 families of mayflies has been recorded in the Mongolian Altai region. Study material was entirely imagos and collected from 78 sites during expeditions led by the Mongolian Aquatic Insect Survey in 2008, 2009 and 2010. Raptobaetopus tenellus, Caenis luctuosa and C. rivulorum are recorded as new to the fauna of Mongolia, and there are new distribution records for Ameletus montanus, Baetis (Acentrella lapponica, Baetis sibiricus, Baetis (Labiobaetis attrebatinus, Centroptilum luteolum, Procloeon pennulatum, Ephemerella aurivillii, Serratella setigera, Ephemera sachalinensis, Ecdyonurus (Afronurus abracadabrus, Cinygmula kurenzovi, Ecdyonurus (Afghanurus vicinus and Epeorus (Belovius pellucidus from the Mongolian Altai region. Baetis vernus and Ephemerella aurivillii are the most frequently encountered species in this region.

  7. WESTERN ESOTERICISM AS AN OBJECT OF RELIGIOUS STUDIES RESEARCH

    Directory of Open Access Journals (Sweden)

    Pavel Nosachev

    2013-06-01

    Full Text Available This article presents a short excursus into the history of research on western esotericism. At the beginning the basic terms esoterica and esotericism are defi ned. Then the history of societies and academic chairs devoted to research on the subject is examined. The source for research on the subject is the Eranos Circle. Later authors such as Frances Yates took up the study which was continued and developed in the volumes authored by Antoine Faivre and Wouter Hanegraaff. Resorting to generalities in describing the various branches of thought on the subject, the author defi nes three main schools: the American School, the Old European School, and finally the New European School. The principle characteristics of the American School is the acknowledgement of the reality of esoteric experience and a tendency to override the border dividing the researcher and the phenomenon researched as well as attempting to popularize the subject in wide sectors of society. The Old European School is characterized by its extremely critical attitude to esotericism, seeing in it little more than a deviation on the path of the development of culture and religion. The New European School, on the other hand, is guided by a post-modern approach to research on this subject, which leads to the constant study of the problem of dialogue and demands a pluralistic attitude in regard to various forms of esotericism. Another characteristic of this school is the intention to include the study of esotericism as an accepted academic discipline. Some basic problems which face the contemporary researcher of this topic are studied by the author separately. The idea of combining the study of esotericism with the study of theology is also touched upon, pointing out the basic contradiction and incompatibility separating the two subjects. Two approaches for Christian researchers are defined — the approach of the tree and the modernistic approach

  8. Cytological studies of Brassicaceae burn. (Cruciferae juss.) from Western Himalayas.

    Science.gov (United States)

    Jeelani, S M; Rani, S; Kumar, S; Kumari, S; Gupta, R C

    2013-01-01

    Cytological studies have been carried out on 12 species of Brassicaceae Burn. on population basis from different geographical areas of Kashmir and Himachal Pradesh in the Western Himalayas. Variable chromosome reports for Barbaraea intermedia (n = 16), Cardamine loxostemonoides (n = 8), Nasturtium officinale (n = 8), Sisymbrium orientale (n = 14) on world-wide basis have been added to the previous reports of these species. The chromosome numbers in seven species as Barbaraea intermedia (n = 8), B. vulgaris (n = 8), Capsella bursa-pastoris (n = 8), Descuriania sophia (n = 10), Rorippa islandica (n = 8), Sisymbrium strictum (n = 7) and Thlaspi alpestre (n = 7) have been worked out for the first time from India. The meiotic course in the populations of seven species such as Barbaraea intermedia, Capsella bursa-pastoris, Coronopus didymus, Descuriania sophia, Nasturtium officinale, Sisymbrium orientale and S. strictum varies from normal to abnormal while all the populations of two species Barbaraea vulgaris and Sisymbrium irio show abnormal meiotic course. Meiotic abnormalities are in the form of cytomixis, chromosomal stickiness, unoriented bivalents, inter-bivalent connections, formation of laggards and bridges, all resulting into abnormal microsporogenesis. Heterogenous sized fertile pollen grains and reduced reproductive potentialities have invariably been observed in all the meiotically abnormal populations. However, the meiotic course in all the populations of Cardamine loxostemonoides, Rorippa islandica and Thalspi alpestre is found to be normal with high pollen fertility.

  9. Blunt abdominal aortic injury: a Western Trauma Association multicenter study.

    Science.gov (United States)

    Shalhub, Sherene; Starnes, Benjamin W; Brenner, Megan L; Biffl, Walter L; Azizzadeh, Ali; Inaba, Kenji; Skiada, Dimitra; Zarzaur, Ben; Nawaf, Cayce; Eriksson, Evert A; Fakhry, Samir M; Paul, Jasmeet S; Kaups, Krista L; Ciesla, David J; Todd, S Rob; Seamon, Mark J; Capano-Wehrle, Lisa M; Jurkovich, Gregory J; Kozar, Rosemary A

    2014-12-01

    Blunt abdominal aortic injury (BAAI) is a rare injury. The objective of the current study was to examine the presentation and management of BAAI at a multi-institutional level. The Western Trauma Association Multi-Center Trials conducted a study of BAAI from 1996 to 2011. Data collected included demographics, injury mechanism, associated injuries, interventions, and complications. Of 392,315 blunt trauma patients, 113 (0.03%) presented with BAAI at 12 major trauma centers (67% male; median age, 38 years; range, 6-88; median Injury Severity Score [ISS], 34; range, 16-75). The leading cause of injury was motor vehicle collisions (60%). Hypotension was documented in 47% of the cases. The most commonly associated injuries were spine fractures (44%) and pneumothorax/hemothorax (42%). Solid organ, small bowel, and large bowel injuries occurred in 38%, 35%, and 28% respectively. BAAI presented as free aortic rupture (32%), pseudoaneurysm (16%), and injuries without aortic external contour abnormality on computed tomography such as large intimal flaps (34%) or intimal tears (18%). Open and endovascular repairs were undertaken as first-choice therapy in 43% and 15% of cases, respectively. Choice of management varied by type of BAAI: 89% of intimal tears were managed nonoperatively, and 96% of aortic ruptures were treated with open repair. Overall mortality was 39%, the majority (68%) occurring in the first 24 hours because of hemorrhage or cardiac arrest. The highest mortality was associated with Zone II aortic ruptures (92%). Follow-up was documented in 38% of live discharges. This is the largest BAAI series reported to date. BAAI presents as a spectrum of injury ranging from minimal aortic injury to aortic rupture. Nonoperative management is successful in uncomplicated cases without external aortic contour abnormality on computed tomography. Highest mortality occurred in free aortic ruptures, suggesting that alternative measures of early noncompressible torso hemorrhage

  10. Assessment of the radiological impact of coal utilization. I. Preliminary studies on Western coal. [Western USA; radionuclide impurities

    Energy Technology Data Exchange (ETDEWEB)

    Styron, C.E.; Casella, V.R.; Farmer, B.M.; Hopkins, L.C.; Jenkins, P.H.; Phillips, C.A.; Robinson, B.

    1979-02-12

    It was the purpose of this project to broadly survey pathways of radionuclides in the coal fuel cycle, identifying critical questions and providing direction for subsequent, definitive studies of radionuclides in coal and the technological enhancement of concentrations of naturally occurring radionuclides through use of this coal. Western coal was selected for study because of its prominence in the National Energy Plan. Specific objectives were to: investigate the fate of radionuclides in the coal fuel cycle and, where possible, delineate the scope of the potential environmental and human health problem associated with radionuclides in coal; compile and evaluate data for uranium-238, uranium-234, lead-210, polonium-210, and thorium-230 in coal; study the release, fate, and accumulation of radionuclides from a power plant burning Western coal; and assess the possible need for additional control technology or standards. Results indicate that concentration of radionuclides in coal from Western mines varies widely, both within and between mines similar to other minor and trace constituents. The concentration of radionuclides in Western coal being mined today is roughly comparable to that of Eastern coal and is slightly below the average for all U.S. coal. A radionuclide balance for a power plant indicated that over 80% of uranium-234, uranium-238, and lead-210 present in the feed coal remained with the fly ash. Thus, collection of fly ash by electrostatic precipitators is of value in controlling radionuclide emissions. An additional 8 to 9% of the radionuclidesremained with bottom ash. Since over 90% of uranium-234, uranium-238, and lead-210 in feed coal remains with bottom ash and fly ash, it is suggested that the possible migration of radionuclides--particularly radon-222 which decays to lead-210 and polonium-210--from the ash pond, warrants further study.

  11. Study of lone working magnetic resonance technologists in Western Australia

    Directory of Open Access Journals (Sweden)

    Tracy Anne Dewland

    2013-12-01

    Full Text Available Objectives: It is recommended that magnetic resonance (MR technologists should not work alone due to potential occupational health risks although lone working is legally acceptable. The objective of this study was to investigate the current situation of lone working MR technologists in Western Australia (WA and any issue against the regulations. Materials and Methods: A questionnaire regarding the issues of occupational health of lone working MR technologists was developed based on relevant literature and distributed to WA MR technologists. Descriptive (percentage of frequency, mean and standard deviation and inferential statistics (Fisher's exact, Chi2 and t tests, and analysis of variance were used to analyze the responses of the yes/no, multiple choice and 5 pt scale questions from the returned questionnaires. Results: The questionnaire response rate was 65.6% (59/90. It was found that about half of the MR technologists (45.8%, 27/59 experienced lone working. The private magnetic resonance imaging (MRI centers were more likely to arrange technologists to work alone (p < 0.05. The respondents expressed positive views on issues of adequacy of training and arrangement, confidence and comfort towards lone working except immediate assistance for emergency (mean: 3. Factors of existence of MRI safety officer (p < 0.05 and nature of lone working (p < 0.001-0.05 affected MR technologists' concerns. Conclusions: Lone working of MR technologists is common in WA especially in private centers. The training and arrangement provided seem to be adequate for meeting the legal requirements. However, several areas should be improved by the workplaces including enhancement on immediate emergency assistance and concern relief.

  12. [Study on the vaccine against dental caries of recombinant Streptococcus lactis and artificial active immunization. V. Extraction, purification and dot blot analysis of RNA in the recombinant S. lactis].

    Science.gov (United States)

    Ling, J; Fan, M

    1997-11-01

    To investigate the transcription level of the pac gene in the recombinant S. lactis, the amount of PAc-specific mRNA was measured by RNA dot blot analysis with the biotin-labeled 1.5 kb PstI fragment of the pac gene. RNA were extracted with guanidinium thiocyanate followed by centrifugation in cesium chloride solutions. These results indicated the sequence of PAc-mRNA in S. lactis HL107 or HL45 was the same as that in S. mutans Ingbritt and showed similar density photograph. The PAc-specific mRNA plated an intermediary role of protein synthesis in recombinant S. lactis and made it complete the expression of cloned pac gene.

  13. Reclassification to the NCAA Division I Football Bowl Subdivision: A Case Study at Western Kentucky University

    Science.gov (United States)

    Upright, Paula A.

    2009-01-01

    The purpose of this study was to describe the reclassification process of Western Kentucky University's football program from the Football Championship Subdivision (FCS) to the Football Bowl Subdivision (FBS), the highest and most visible level of NCAA competition. Three research questions guided the study: (a) Why did Western Kentucky University…

  14. The Problem of World Order in Western IR Studies

    Directory of Open Access Journals (Sweden)

    Maria Victorovna Soljanova

    2016-01-01

    Full Text Available The article "Problem of world order in modern Western studies" is the study of one of the most debated issues in the science of international relations - world order. Discussion of the structure of world order is underway in various countries, both at the state level and in the expert community. Some researchers insist on the fact that after the end of the cold war, the collapse of the bipolar model of international relations, the world has become unipolar. Others argue that the increase in the number of centers of power and the need for a multilateral approach to solving global problems (terrorism, proliferation of weapons of mass destruction, environmental and climate issues talking about the formation of multipolarity. However, it should be recognized that currently no widely accepted theoretical and conceptual apparatus, which complicates not only the study of the world order, but makes it impossible to search for common approaches of the international community in solving the problems associated with global development, new challenges and threats. The author of this article seeks to research and analyze the various theoretical paradigms (neo-realism, neo-liberalism, institutionalism, neo-marxism, etc. and concepts to form a coherent picture of the structure of the world system, its main features and to offer readers the vision of the concept of "world order". Thus, the article notes that the multidimensional structure of the modern system of international relations established after the end of the cold war is so complex that none of the concepts can claim to accurate interpretation of the world order. The modern system differs from systems of the past centuries. Characteristics inherent in it (on the one hand, the increasing global processes in economy, politics, culture, etc., on the other, the attraction to return to the concept of "nation state", the closure of borders, the disintegration, require new approaches to the study of world

  15. Empathy and Emotional Intelligence among Eastern and Western Counsellor Trainees: A Preliminary Study

    Science.gov (United States)

    Young Kaelber, Kara A.; Schwartz, Robert C.

    2014-01-01

    This study explored degree of empathy and emotional intelligence among Thai (n?=?48) and American (n?=?53) counsellor trainees to determine if differences in Eastern and Western cultural orientations (e.g., interdependent versus independent self-construals) affect foundational counselling skills. Results indicated that Western trainees showed…

  16. Fast and Simple micro-RNA Northern Blots

    Directory of Open Access Journals (Sweden)

    Nham Tran

    2009-01-01

    Full Text Available RNA northern blots provide robust measurements of gene expression. The simple northern blot technique described in this report has been optimised to provide rapid, reproducible detection and analysis of mature and precursor forms of microRNAs. This protocol economises on the use of commercially available components and secondly reduces the hybridisation step to 2 hours.

  17. Larval connectivity studies in the Western Iberian Peninsula

    Science.gov (United States)

    Dubert, Jesus; Nolasco, Rita; Queiroga, Henrique

    2010-05-01

    The study of the connectivity between populations is one of the 'hot' applications of numerical models of the ocean circulation. An IBM (Individual Based model) was developed, using Carcinus manenas larvae crab as a model. A set of particles was used as a representation of larvae, in order to study their larval life cycle, including the larval growth, larval mortality (both depending on temperature and salinity), larval dispersal by currents, diel vertical migration, and larval recruitment. The life cycle of every larvae in the ocean, was modeled from zoeae 1 stage to megalopae stage, during typical periods of 30-50 days. Larvae were initialized in 14 estuarine systems of the Atlantic Western Iberian Peninsula, from January to July. In every period, a number of 225 larvae are initialized in everyone of the 14 considered estuaries, with fortynighly periodicity. The larvae evolves during the (variable, depending mainly on temperature) period of growth in the ocean, and when a larvae reach the age for recruit, if it is located in the neighborhood of the considered estuarine systems, the larvae is accounted as a recruited larvae in that place. With this methodology, a connectivity matrix can be computed, acconting for the 225 larvae emitted in every estuary, the number of larvae that reaches the every place. The connectivity matrix depends strongly on the current regime along the Atlantic coast of Iberian Peninsula, and has been calculated for the present circulation, for the period 2001 to 2009, for runs with realistic forcing with NCEP2 and Quikscat (for winds) forcing. The connectivity matrix, have also been calculated for climatological runs. For the present climatological conditions, it is observed the prevalence of southward transport for the period January-July, because the prevalence of Northerly winds along the west coast of IP in the COADS present time climatology. Strong dispersal is observed at the Northern estuaries, during winter with strong loss of

  18. Ethnic Studies in the United States as decolonial studies within the overall university system westernized

    Directory of Open Access Journals (Sweden)

    Ramón Grosfoguel

    2013-01-01

    Full Text Available This article is an analysis of the Westernized university and its Eurocentric fundamentalism in relation to the subaltern struggles of racialized groups in the United States and its impact on the formation of ethnic studies in the university’s epistemic structure. The article goes on to discuss questions of epistemic racism/sexism and the dilemmas that ethnic studies programs confront today in particular forms of disciplinary colonization, liberal multiculturalism and identity politics.

  19. 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.

  20. Study of geothermal prospects in the western United States

    Energy Technology Data Exchange (ETDEWEB)

    1976-08-20

    The commercial development potential of 13 underdeveloped geothermal prospects in the Western United States has been examined and the prospects have been ranked in order of relative potential for development on the basis of investment considerations. The following were considered in the ranking: geotechnical and engineering data, energy market accessibility, administrative constraints, and environmental and socio-economic factors. The primary ranking criterion is the unit cost of energy production expected from each prospect. This criterion is obtained principally from expected reservoir temperatures and depths. Secondary criteria are administrative constraints, environmental factors and the quality of the geotechnical data. The Roosevelt, Utah, prospect ranks first in development potential followed in order by Beowawe, Nevada; Coso Hot Springs, California; Long Valley, California; and Brady's Hot Springs, Nevada.

  1. Study of geothermal prospects in the western United States

    Energy Technology Data Exchange (ETDEWEB)

    1975-01-01

    The commercial development potential of 13 underdeveloped geothermal prospects in the western United States has been examined and the prospects have been ranked in order of relative potential for development on the basis of investment considerations. The following were considered in the ranking: geotechnical and engineering data, energy market accessibility, administrative constraints, and environmental and socio-economic factors. The primary ranking criterion is the unit cost of energy production expected from each prospect. This criterion is obtained principally from expected reservoir temperatures and depths. Secondary criteria are administrative constraints, environmental factors and the quality of the geotechnical data. The Roosevelt, Utah, prospect ranks first in development potential followed in order by Beowawe, Nevada; Coso Hot Springs, California; Long Valley, California; and Brady's Hot Springs, Nevada.

  2. 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

    Fresh biopsy material for molecular biological investigations is not obtainable from all relevant normal human tissues. We studied the feasibility of using RNA and DNA from autopsies for Northern and Southern blot analysis. Tissue samples from seven organs were obtained from 10 autopsies performed...... 21-118 h postmortem. Extracted RNA and DNA were examined by Northern and Southern blot analysis using oligo-labelled human DNA probes recognizing gene transcripts of 2-5 kb. The results indicated that, in general, Northern blot analysis was feasible with the applied probes when the tissue...... 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...

  3. Reliability of Blotting Techniques to Assess Contact Lens Water Content.

    Science.gov (United States)

    Cañadas, Pilar; López-Miguel, Alberto; Gómez, Alba; López-de la Rosa, Alberto; Fernández, Itziar; González-García, María J

    2018-02-15

    To determine the reliability of wet and modified dry blotting techniques used in the gravimetric method to assess contact lens (CL) water content (WC), the accuracy of both techniques in comparison with the nominal WC, and also their agreement. We evaluated hydrated and dry CL mass values and WC using the gravimetric method in 440 daily disposable CLs. Samples assessed corresponded to Dailies Total 1, Dailies AquaComfort Plus, 1-Day Acuvue TruEye, and Biotrue ONEday. Back vertex power ranged from +3.00 diopters (D) to -6.00 D. Within-subject coefficient of variation (CVw) and intraclass correlation coefficients were calculated. Bland-Altman analysis was also performed. The modified dry blotting technique yielded significantly (P≤0.0001) higher hydrated CL mass values. The wet blotting technique provided significantly (P≤0.04) better consistency than the modified dry one. Values of CVw for wet and modified dry blotting techniques ranged from 1.2% to 2.1% and from 3.7% to 5.4%, respectively. As for dry CL mass values, CVw values were not significantly different (P≥0.05) between wet (range: 1.1%-1.9%) and dry (range: 1.0%-5.1%) blotting techniques, except for Dailies AquaComfort Plus (P=0.03). Bland-Altman analysis showed poor agreement between the techniques. The wet blotting technique yielded WC values close (around 1%) to nominal ones, in contrast to modified dry blotting technique (≥2.5%). The wet blotting technique is not only more reliable than the modified dry one when obtaining hydrated CL mass but also provides more accurate nominal WC measurements. Agreement between the techniques was poor.

  4. Western U.S.-Canada crossborder case study : activity 2 : task D : conduct regional and local trucking case studies

    Science.gov (United States)

    1995-12-01

    This case study examines trucking across the western U.S.-Canada border and how it is influenced by truck size and weight (TS&W) regulations. Western border trucking differs from eastern border trucking in terms of the types of commodities being hand...

  5. Morphometric study of cricoid cartilages in Western India

    Directory of Open Access Journals (Sweden)

    Mohini Joshi

    2011-10-01

    Full Text Available BackgroundIt is important to determine the size and proportion of thelarynx as such information is useful in procedures such asintubation, endoscopy and surgical manipulations. Recentinterest in the cases of subglottic stenosis and postintubationalstenosis of the lower respiratory tract has ledto renewed interest in ascertaining the measurements ofthe various laryngeal cartilages. The aim of the presentstudy was to collect morphometric data of cricoid cartilagefrom a regional population.MethodFifty laryngeal preparations from adult cadavers of WesternIndia were assessed. Sections were prepared via dissectionand the removed cricoid cartilages then measured andweighed.ResultsThe mean antero-posterior diameter (19.29±2.47 of thecricoid cartilage was greater than the average transversediameter (18.33±2.26. The height of arch of cricoidcartilage was 6.54±1.23mm and height of lamina was21.45±1.97mm. Mean weight of cricoid cartilage was4.53±1.27grams. The shape of the cricoid cartilage wasovoid in 46% of cases, oval in 38%, pear shaped in 12% andnarrow-oblong in 4% of cases.ConclusionInter-subject variability in the dimensions of cricoidcartilages was observed. The large difference in almost allsizes and shapes of the cricoid cartilage makes it difficult tostandardise the rigid stents used in these organs.Endotracheal tubes of the appropriate size should thereforebe based on the measurements of individual patients.Clinicians should therefore be aware of morphologicalvariations as they are of fundamental clinical importance.Key WordsCricoid cartilage, larynx, morphometry

  6. WESTERN STUDIES OF THE QUR’ANIC NARRATIVE: from the Historical Orientation into the Literary Analysis

    Directory of Open Access Journals (Sweden)

    Munirul Ikhwan

    2010-08-01

    Full Text Available The beginnings of Western interest in the Qur’an can be traced back to the appearance of the first complete translation of the Qur’an into Latin by Robert of Ketton in the twelfth century when the Muslim and Western Christian worlds has begun a long-running confrontation. In the eighteenth century, Western scholars began to be interested in studying the history and sources of the Qur’an. The Qur’anic narrative, which has its parallels in the Judeo-Christian traditions, has been studied from the historical perspective. In this approach, everything in the Qur’an that can be also found in earlier scriptures, is considered as borrowed, and every story that the Qur’an modifies is viewed as distorted. Recent Western studies have shifted into a new arena, studying the contents and styles of the Qur’anic narrative by analyzing its discourse and narrativity. Keywords: Western scholarship, Qur’anic studies, Judeo-Christian source

  7. Western Wind and Solar Integration Study: Executive Summary, May 2010

    Energy Technology Data Exchange (ETDEWEB)

    Piwko, R.; Clark, K.; Freeman, L.; Jordan, G.; Miller, N.

    2010-05-01

    This Study investigates the operational impact of up to 35% energy penetration of wind, photovoltaics (PVs), and concentrating solar power (CSP) on the power system operated by the WestConnect group of utilities in Arizona, Colorado, Nevada, New Mexico, and Wyoming.

  8. Small-scale anaerobic digestion: case studies in Western Europe

    OpenAIRE

    De Dobbelaere, Anke; De Keulenaere, Bram; De Mey, Jonathan; Lebuf, Viooltje; Meers, Erik; Ryckaert, Bart; Schollier, Céline; Van Driessche, Jason

    2015-01-01

    This brochure is intended to meet farmers’ demand for more information concerning the current market situation of the small-scale anaerobic digestion technology. The authors aim to quickly guide those who are interested in the technology so that they can adopt a targeted approach in their search for information. This brochure elaborates on how the implementation of small-scale anaerobic digestion can take place at the company level, based on five practical examples. A broad market study in...

  9. Geogenic Groundwater Contamination: A Case Study Of Canakkale - Western Turkey

    Science.gov (United States)

    Deniz, Ozan; Çalık, Ayten

    2016-04-01

    Study area is located NW of Turkey. Total area of the drainage basin is 465 square kilometers and mostly covered by volcanic rocks. Majority of these rocks have highly altered and lost their primary properties because of alteration processes. Especially argillic alteration is common. Tectonic movements and cooling fractures were created suitable circulation environment of groundwater in the rocks (secondary porosity). Alteration affects the composition of groundwater and some rock elements pass into groundwater during the movement of water in the cavities of rocks. High concentration of natural contaminants related to water-rock interaction in spring water has been studied in this research. Field measurements such as pH, electrical conductivity, temperature, oxidation-reduction potential and salinity carried out in 500 water points (spring, drilling, well and stream). 150 water samples taken from the water points and 50 rock samples taken from the source of springs has been investigated in point of major anion-cations, heavy metals and trace elements. Some components in the water such as pH (3.5-9.1), specific electrical conductivity (84-6400 microS/cm), aluminum (27-44902 ppb), iron (10-8048 ppb), manganese (0.13-8740 ppb), nickel (0.2-627 ppb), lead (0.1-42.5 ppb) and sulphate (10 to 1940 ppm) extremely high or low in the springs sourced from especially highly altered Miocene aged volcanic rocks. Some measured parameters highly above according to European Communities Drinking Water Regulations (2007) and TS266 (2015-Intended for Human Consumption Water Regulations of Turkey) drinking water standards. The most common element which is found in the groundwater is aluminum that is higher than to the drinking water standards (200 microg/L). The highest levels of the Al values measured in acidic waters with very low pH (3.4) emerging from altered volcanic rocks because of acid mine drainage in Obakoy district, north of the study area. The abundance of this element in

  10. Western Perceptions of Hong Kong Ten Years on: A Corpus-Driven Critical Discourse Study

    Science.gov (United States)

    Cheng, Winnie; Lam, Phoenix W. Y.

    2013-01-01

    This article studies the Western perceptions of and relations with Hong Kong a decade after the reversion of the sovereignty from Britain to China in 1997. Previous studies have demonstrated that the West had a significantly negative view on the future of Hong Kong with respect to the handover. According to recent observations, however, the…

  11. TSE strain differentiation in mice by immunohistochemical PrPSc profiles and triplex Western blot

    NARCIS (Netherlands)

    Keulen, van L.J.M.; Langeveld, J.P.M.; Dolstra, C.H.; Jacobs, J.G.; Bossers, A.; Zijderveld, van F.G.

    2015-01-01

    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

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

    Science.gov (United States)

    1992-09-16

    months gestation ME.2C Rhogam injection 0-no;1-yes ME.2Ca # mo since Rhogam _ _ ME.2D Abn1 PAP 0-no;I-ABNL ME.2Da Date last abnl (MMYY) ME.2E SP had...acyclovir ME.4Ae Sexual IC c recur 59 ME.4B #z’s Gonorrhea ME.4Ba Date last x ME.4C #*X’ NGU/CZRV _ ME.4Ca Date last x ME.4D #XZS syphilis ME.4Da Date

  13. Day Reporting Center and Recidivism: Comparing Offender Groups in a Western Pennsylvania County Study

    Science.gov (United States)

    Champion, David R.; Harvey, Patrick J.; Schanz, Youngyol Yim

    2011-01-01

    In this study the authors report on an investigation comparing the recidivism and other variables of two similar offender populations in a western Pennsylvania county. The two groups were comparable in offense type, size (N = 63 for each) and other variables such as sex, race and age range. One group represented offenders who received a sentence…

  14. Commercial morel harvesters and buyers in western Montana: an exploratory study of the 2001 harvesting season.

    Science.gov (United States)

    Rebecca J. McLain; Erika Mark McFarlane; Susan J. Alexander

    2005-01-01

    This exploratory study examined aspects of the social organization of the commercial wild morel industry in western Montana during 2001. We talked with 18 key informants (7 buyers and 11 pickers) and observed social interactions at one buying station near the Kootenai National Forest and three buying stations near the Bitterroot National Forest. The key informant and...

  15. Studies on the planktonic Decapoda and Stomatopoda (Crustacea) from the western Bay of Bengal

    Digital Repository Service at National Institute of Oceanography (India)

    Paulinose, V.T.; Nair, V.R.; Goswami, S.C.

    Abundance and distribution of decapods and stomatopods were studied, based on 90 samples collected from the western Bay of Bengal (Lat. 09 degrees 58' and 20 degrees 50'N and long. 80 degrees 00'E) between 24 August and 2 October, 1976. The decapod...

  16. Nursery cultural practices to achieve targets: A case study in western larch irrigation

    Science.gov (United States)

    Anthony S. Davis; Robert F. Keefe

    2011-01-01

    Nursery cultural practices are used to help growers achieve pre-determined size and physiological targets for seedlings. In that regard, irrigation is used to accelerate or slow growth and as a trigger for changing growth phase. In a case study highlighting the effects of irrigation on seedling development, western larch (Larix occidentalis Nutt.) seedlings were grown...

  17. Jarosite occurrence in the Deccan Volcanic Province of Kachchh, western India: Spectroscopic studies on a Martian analog locality

    National Research Council Canada - National Science Library

    Bhattacharya, Satadru; Mitra, Souvik; Gupta, Saibal; Jain, Nirmala; Chauhan, Prakash; Parthasarathy, G; Ajai

    2016-01-01

    .... Here we report an analog terrestrial locality hosting jarosite from Matanumadh, Kachchh, western India, using detailed spectroscopic studies on weathered basalts of the Deccan Volcanic Province...

  18. Development of new staining technology "eastern blotting" using monoclonal antibody.

    Science.gov (United States)

    Morinaga, Osamu; Shoyama, Yukihiro

    2011-03-01

    Ginsenosides contained in Panax species were separated by silica gel TLC blotted to a polyvinylidene difluoride (PVDF) membrane which was dipped in a sodium periodide (NaIO(4)) solution and reacted with protein, preparing a ginsenoside-protein conjugate for binding a ginsenoside on a PVDF membrane. The blotted spots were stained by anti-ginsenoside-Rb1 monoclonal antibody (MAb) and anti-ginsenoside-Rg1MAb, respectively. The newly established immunostaining method, eastern blotting was applied for the determination of ginsenosides possessing protopanaxadiol and/or protopanaxatriol. Double staining of eastern blotting for ginsenosides using anti-ginsenoside-Rb1 MAb and anti-ginsenoside-Rg1 MAb promoted complete identification of ginsenosides in Panax species. This technique has been devised for the chromatographic separation and identification of ginsenosides using polyethersulfone (PES) membrane. It caused an acceptable separation of ginsenoside-Rb1, -Rc and -Rd in various ginseng extracts. Newly developed technique is quite simple and applies for immunoassay system. Ginsenosides separated using a PES membrane were directly treated with a NaIO(4) solution and then reacted with bovine serum albumin (BSA) for making a ginsenoside-protein conjugate. After the blocking, anti-ginsenoside-Rb1 MAb recognized a ginsenoside on a PES membrane and then a sec-ond antibody labeled with enzyme reacted to the first antibody. Finally a substrate was oxidized with the enzyme and de-veloped the staining of ginsenosides. The staining spots of ginsenosides on membrane were quantitatively evaluated by NIH Image indicating at least 62.5 ng of each ginsenoside-Rb1, -Rc and -Rd were detected with clarity. The determination range of three ginsenosides was from 0.125 to 2.0 µg of direct amount on PES membrane.

  19. Seasonal effects on great ape health: a case study of wild chimpanzees and Western gorillas.

    Directory of Open Access Journals (Sweden)

    Shelly Masi

    Full Text Available Among factors affecting animal health, environmental influences may directly or indirectly impact host nutritional condition, fecundity, and their degree of parasitism. Our closest relatives, the great apes, are all endangered and particularly sensitive to infectious diseases. Both chimpanzees and western gorillas experience large seasonal variations in fruit availability but only western gorillas accordingly show large changes in their degree of frugivory. The aim of this study is to investigate and compare factors affecting health (through records of clinical signs, urine, and faecal samples of habituated wild ape populations: a community (N = 46 individuals of chimpanzees (Pan troglodytes in Kanyawara, Kibale National Park (Uganda, and a western gorilla (G. gorilla group (N = 13 in Bai Hokou in the Dzanga-Ndoki National Park (Central African Republic. Ape health monitoring was carried out in the wet and dry seasons (chimpanzees: July-December 2006; gorillas: April-July 2008 and December 2008-February 2009. Compared to chimpanzees, western gorillas were shown to have marginally greater parasite diversity, higher prevalence and intensity of both parasite and urine infections, and lower occurrence of diarrhea and wounds. Parasite infections (prevalence and load, but not abnormal urine parameters, were significantly higher during the dry season of the study period for western gorillas, who thus appeared more affected by the large temporal changes in the environment in comparison to chimpanzees. Infant gorillas were the most susceptible among all the age/sex classes (of both apes having much more intense infections and urine blood concentrations, again during the dry season. Long term studies are needed to confirm the influence of seasonal factors on health and parasitism of these great apes. However, this study suggest climate change and forest fragmentation leading to potentially larger seasonal fluctuations of the environment may affect

  20. Are Parents More Likely to Be Unemployed? A Study of Nine Western Democracies

    OpenAIRE

    Stjärnfäldt, Richard

    2016-01-01

    The relation between parenthood and market work is an established field of study in demography. This thesis specifically focuses on the relation between parenthood and unemployment, the involuntary absence of work, in a cross sectional study of nine western democracies. The studied countries are Canada, France, Germany, Italy, Poland, Spain, Sweden, the United Kingdom, and the United States. A significant relation between motherhood and unemployment where found in Germany, Poland, the United ...

  1. Interactions between traditional Chinese medicine and western drugs in Taiwan: A population-based study.

    Science.gov (United States)

    Chen, Kuan Chen; Lu, Richard; Iqbal, Usman; Hsu, Ko-Ching; Chen, Bi-Li; Nguyen, Phung-Anh; Yang, Hsuan-Chia; Huang, Chih-Wei; Li, Yu-Chuan Jack; Jian, Wen-Shan; Tsai, Shin-Han

    2015-12-01

    Drug-drug interactions have long been an active research area in clinical medicine. In Taiwan, however, the widespread use of traditional Chinese medicines (TCM) presents additional complexity to the topic. Therefore, it is important to see the interaction between traditional Chinese and western medicine. (1) To create a comprehensive database of multi-herb/western drug interactions indexed according to the ways in which physicians actually practice and (2) to measure this database's impact on the detection of adverse effects between traditional Chinese medicine compounds and western medicines. First, a multi-herb/western medicine drug interactions database was created by separating each TCM compound into its constituent herbs. Each individual herb was then checked against an existing single-herb/western drug interactions database. The data source comes from the National Health Insurance research database, which spans the years 1998-2011. This study estimated the interaction prevalence rate and further separated the rates according to patient characteristics, distribution by county, and hospital accreditation levels. Finally, this new database was integrated into a computer order entry module of the electronic medical records system of a regional teaching hospital. The effects it had were measured for two months. The most commonly interacting Chinese herbs were Ephedrae Herba and Angelicae Sinensis Radix/Angelicae Dahuricae Radix. Ephedrae Herba contains active ingredients similar to in ephedrine. 15 kinds of traditional Chinese medicine compounds contain Ephedrae Herba. Angelicae Sinensis Radix and Angelicae Dahuricae Radix contain ingredients similar to coumarin, a blood thinner. 9 kinds of traditional Chinese medicine compounds contained Angelicae Sinensis Radix/Angelicae Dahuricae Radix. In the period from 1998 to 2011, the prevalence of herb-drug interactions related to Ephedrae Herba was 0.18%. The most commonly prescribed traditional Chinese compounds were

  2. NEW PROBLEMATIZATION WAYS OF THE GREAT PATRIOTIC WAR HISTORY IN WESTERN RUSSIAN STUDIES

    Directory of Open Access Journals (Sweden)

    Наталья Валерьевна Трубникова

    2013-12-01

    Full Text Available The article examines the new thematic perspectives prevailing in Western historiography of the Great Patriotic War. The analysis is implemented on the basis of the French scientific periodicals of 2001–2013 years. Historical survey of contemporary Russian Studies seeking to fill research gaps and form a new problem fields in the study of the World War II Eastern Front history. These studies continue the trend of the second half of 1990 – the beginning of the 2000s, which withdrow themes of Soviet history from politicized totalitarian interpretations to ideologically neutral field of renewed social history. New vectors of study based on the use of newly opened archives hold  a "context" study of pre–war and post–war international political events related to the activities of the Soviet regime, including sensitive issues of the Soviet Union geopolitical expansion in Eastern Europe; the survey of different social ( ethnic, gender , religious groups in the war and in the " out of the war " process; Stalins's strategies of management, military repression, " purges " and ideological influence on the population.Purpose: To reveal the areas of contemporary Western Russian Studies research in the history of the Great Patriotic War.Methodology : problem historiography.Results: the newest thematic areas of research and methodological techniques of WWII Western Russien studies are identified.Practical implications: research and teaching activities in the field of the Russian history.DOI: http://dx.doi.org/10.12731/2218-7405-2013-10-26

  3. Detection of M2 antimitochondrial antibodies by dot blot assay is more specific than by enzyme linked immunosorbent assay.

    Science.gov (United States)

    Bargou, I; Mankaï, A; Jamaa, A; Ben Jazia, I; Skandrani, K; Sfar, H; Baccouche, A; Ajmi, S; Letaief, A; Fabien, N; Jeddi, M; Ghedira, I

    2008-02-01

    The objective of our study was, in one hand, to determine the sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of ELISA and dot blot assay to investigate IgG M2 antimitochondrial antibodies (M2 AMA) and, on the other hand, to compare these results with those of indirect immunofluorescence technique (IIF). Sera from patients suffering from primary biliary cirrhosis (PBC) (n=55), systemic lupus erythematosus (n=21), celiac disease (n=30) and blood donors (n=75) were analyzed. M2 AMA were detected by ELISA and dot blot using pyruvate dehydrogenase purified from porcine heart and by IIF on cryostat sections of rat liver-kidney-stomach. IIF was more sensitive (98%) than ELISA (93%) and dot blot (91%). The specificity of AMA for PBC using IIF, ELISA and dot blot reached 100%, 92% and 100%, respectively. The PPV of IIF, ELISA and dot blot was 100%, 93% and 100%, respectively. The NPV was 98% for IIF, 92% for ELISA and 91% for dot blot. Dot blot, using purified pyruvate dehydrogenase, had a higher specificity than ELISA and may be useful in confirming the specificity of AMA in cases of doubt with IIF.

  4. [The trend and prospect of studies on the history of Western medicine in Korea.].

    Science.gov (United States)

    Kim, Ock Joo

    2010-06-30

    Studies on the history of Western medicine in Korea began to be actively conducted and published since the restart of the Korean Society for the History of Medicine in 1991, which had been originally inaugurated in 1947, and the publication of its official journal, the Korean Journal of Medical History in 1992. In 1970s and 1980s, even before the start of the Journal, articles on a history of Western medicine were published mainly written by physicians in medical journals. This paper aims to provide an overview of the publications on the history of Western medicine in Korea, comparing papers published in the Journal with those published in other journals. Authors of the papers in the Journal are those who majored in history of medicine or history science whose initial educational backgrounds were medicine or science, whereas authors of the papers in other journals majored in Western history, economic history, social history, religious history, or women's history. While a large portion of papers in the Journal deal with medicine in ancient Greek or in modern America with no paper on medieval medicine, the papers in other journals deal with more various periods including ancient, medieval and modern periods and with diverse areas including France, Britain, Germany, Europe etc. Recent trends in 2000s show an increase in the number of researchers who published the history of Western medicine in other journals, total number of their publications, and the topics that they dealt with in their papers. In contrast, however, the number of researchers published in the Journal, the number of the papers and its topics - all decreased in recent years. Only three papers on the history of diseases have been published in the Journal, while eleven published in other journals. In order to stimulate research on the history of Western medicine in Korea, concerted efforts are necessary including academic communication among various disciplines, formulation of a long term plan to enlarge

  5. Use of a Combined Duplex PCR/Dot Blot Assay for more sensitive genetic characterization

    Directory of Open Access Journals (Sweden)

    Erin Curry MS

    2008-01-01

    Full Text Available A reliable and sensitive method of genetic analysis is necessary to detect multiple specific nucleic acid sequences from samples containing limited template. The most widely utilized method of specific gene detection, polymerase chain reaction (PCR, imparts inconsistent results when assessing samples with restricted template, especially in a multiplex reaction when copies of target genes are unequal. This study aimed to compare two methods of PCR product analysis, fluorescent detection following agarose gel electrophoresis or dot blot hybridization with chemiluminescent evaluation, in the detection of a single copy gene (SRY and a multicopy gene (β-actin. Bovine embryo sex determination was employed to exploit the limited DNA template available and the target genes of unequal copies. Primers were used either independently or together in a duplex reaction with purified bovine genomic DNA or DNA isolated from embryos. When used independently, SRY and β-actin products were detected on a gel at the equivalent of 4-cell or 1-cell of DNA, respectively; however, the duplex reaction produced visible SRY bands at the 256 cell DNA equivalent and β-actin products at the 64 cell DNA equivalent. Upon blotting and hybridization of the duplex PCR reaction, product was visible at the 1–4 cell DNA equivalent. Duplex PCR was also conducted on 186 bovine embryos and product was subjected to gel electrophoresis or dot-blot hybridization in duplicate. Using PCR alone, sex determination was not possible for 22.6% of the samples. Using PCR combined with dot blot hybridization, 100.0% of the samples exhibited either both the male specific and β-actin products or the β-actin signal alone, indicating that the reaction worked in all samples. This study demonstrated that PCR amplification followed by dot blot hybridization provided more conclusive results in the evaluation of samples with low DNA concentrations and target genes of unequal copies.

  6. Cardiometabolic correlates of low type 2 diabetes incidence in western Alaska Native people - The WATCH study.

    Science.gov (United States)

    Koller, Kathryn R; Metzger, Jesse S; Jolly, Stacey E; Umans, Jason G; Hopkins, Scarlett E; Kaufmann, Cristiane; Wilson, Amy S; Ebbesson, Sven O E; Raymer, Terry W; Austin, Melissa A; Howard, Barbara V; Boyer, Bert B

    2015-06-01

    Previously rare among Alaska Native (AN) people, type 2 diabetes (DM2) prevalence as indicated by registry data has increased by as much as 300% in some western Alaska regions. We sought to determine prevalence and incidence of DM2 and analyze associated cardiometabolic risk factors in western AN people. DM2 and prediabetes prevalence and incidence were determined by the Western Alaska Tribal Collaborative for Health using consolidated data from cohort studies conducted during 2000-2010. Crude and age-adjusted incidence for DM2 and prediabetes were calculated using 2010 American Diabetes Association criteria. Effects of covariates on DM2 and prediabetes were determined using univariate and multivariate Cox proportional hazards analyses, adjusted for age and sex. Excluding baseline diabetes (n=124, 4.5%), 53 cases of new DM2 were identified among 2630 participants. Age- and sex-adjusted DM2 incidence was 4.3/1000 (95% CI 2.9, 5.0) person-years over an average 5.9-year follow up. After excluding baseline prediabetes, 387 new cases of prediabetes were identified among 1841 participants; adjusted prediabetes incidence was 44.5/1000 (95% CI 39.5, 49.5) person years. Independent predictors for DM2 included age, impaired fasting glucose, and metabolic syndrome; family history of diabetes and obesity were additional independent predictors for prediabetes. DM2 incidence in western AN people is substantially lower than that for U.S. whites; however, incidence of prediabetes is more than 10-fold higher than western AN DM2 incidence and more closely aligned with U.S. rates. Interventions aimed at achieving healthy lifestyles are needed to minimize risk factors and maximize protective factors for DM2 in this population. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  7. Quantitative dot blot analysis (QDB), a versatile high throughput immunoblot method.

    Science.gov (United States)

    Tian, Geng; Tang, Fangrong; Yang, Chunhua; Zhang, Wenfeng; Bergquist, Jonas; Wang, Bin; Mi, Jia; Zhang, Jiandi

    2017-08-29

    Lacking access to an affordable method of high throughput immunoblot analysis for daily use remains a big challenge for scientists worldwide. We proposed here Quantitative Dot Blot analysis (QDB) to meet this demand. With the defined linear range, QDB analysis fundamentally transforms traditional immunoblot method into a true quantitative assay. Its convenience in analyzing large number of samples also enables bench scientists to examine protein expression levels from multiple parameters. In addition, the small amount of sample lysates needed for analysis means significant saving in research sources and efforts. This method was evaluated at both cellular and tissue levels with unexpected observations otherwise would be hard to achieve using conventional immunoblot methods like Western blot analysis. Using QDB technique, we were able to observed an age-dependent significant alteration of CAPG protein expression level in TRAMP mice. We believe that the adoption of QDB analysis would have immediate impact on biological and biomedical research to provide much needed high-throughput information at protein level in this "Big Data" era.

  8. CONTENT ANALYSIS OF AN EXPLORATORY STUDY REGARDING THE COMMUNICATION OF LOCAL ADMINISTRATION IN WESTERN ROMANIA

    OpenAIRE

    Florina Maria Bente; Corneliu Bente

    2013-01-01

    The purpose of this paper was to emphasize the main characteristics of communication in Western Romanian administration. Using qualitative research we investigated citizens' opinions on the communication in local public administration. As an exploratory study we performed a content analysis of information gathered from five focusgroups. By analyzing data it was possible to highlight the aspects that please or discontent the citizens. The goal was to collect a series of essential events from w...

  9. [Microscopic identification study of western herbs. Part I: Three Echinacea-species roots].

    Science.gov (United States)

    Li, Qin; Chu, Chu; Tan, Zhenfeng; Wang, Yaqiong; Wei, Jiafu; Li, Ping; Zhao, Zhongzhen

    2009-11-01

    To make microscopic identification research of three Echinacea-species roots recorded in the United States Pharmacopeia. The root transverse section and powder of E. angustifolia, E. pallida, and E. purpurea were observed. The main microscopic features were photographed. The main microscopic features of transverse section and powder in three Echinacea-species roots are basically similar, except for some diagnostic differences. The results provide reliable reference for the authentication of raw materials of western herbal studies.

  10. Maxillofacial Fractures: Its features and Occurrence in Western Uttar Pradesh,India- A Retrospective Study.

    OpenAIRE

    Dr. Swapnil S. Bumb; Dr. S K Jain; Dr. A K Chaudhary; Dr. Sadakat Ali

    2013-01-01

    Introduction: The successful surgical treatment of maxillofacial fractures consists of early recognition of fractured site, etiologic factors and demographic patterns. In Western Uttar Pradesh, Road Traffic Accidents are leading cause of maxillofacial fractures followed by facial assault. Mandibular fractures followed by upper face fractures are the leading causes of maxillofacial fractures. Aims and Objectives: The aim of this retrospective study is to investigate the pattern of maxillo...

  11. Leveraging and Integrating Eastern and Western Insights for Human Engagement Studies in HCI

    DEFF Research Database (Denmark)

    Law, Effie Lai-Chong; Silpasuwanchai, Chaklam; Ren, Xiangshi

    2015-01-01

    Human engagement is at the heart of every interactive technology. However, a concrete framework for synergizing the capabilities of humans and technologies to allow fully engaging interactions to happen is yet to be developed. We posit that such a framework should be grounded in a deeper understa...... in their environments, including technological artefacts. Discussions of leveraging and possibly integrating Eastern and Western insights for human engagement studies will be an exciting and a radical forum for the HCI community....

  12. A case study of Megaselia scalaris (Diptera: Phoridae) causing ocular myiasis in a western hognose snake.

    Science.gov (United States)

    Diclaro, J W; Lehnert, M S; Mitola, M A; Pereira, R M; Koehler, P G

    2011-07-01

    Late instar larvae of the scuttle fly, Megaselia scalaris Loew, were found near the right eye of a live captive-reared western hognose snake, Heterodon nasicus Baird and Girard. Dissection and removal of the snake's dorsal cranial bones revealed tissue degradation of the infected eye, the optic nerve, and the brain case; we suggest that these factors contributed to the death of this snake. This case study further demonstrates the opportunistic behavior of M. scalaris.

  13. Breast feeding initiation rate across Western countries: does religion matter? An ecological study.

    Science.gov (United States)

    Bernard, Jonathan Y; Cohen, Emmanuel; Kramer, Michael S

    2016-01-01

    Breast feeding initiation rates remain below 80% in some Western countries. Many individual-level determinants are known; however, less is known regarding cultural and societal determinants, such as religion, that could explain population-level variations. We examined the correlations of the proportions of Catholics and Protestants with the breast feeding initiation rates across and within Western countries. Using publicly available data, we carried out an ecological study comparing the proportions of Catholics and Protestants with the rates of breast feeding initiation. We correlated data at the country level, and additionally explored within-country data in five Western countries: France (Departments), Ireland (counties), the UK (countries), Canada (provinces) and the USA (states). Our analyses accounted for human development index, gross domestic product and population density. We observed a negative correlation (r=-0.30) between the proportion of Catholics and the rate of breast feeding initiation in Western countries. This correlation was consistent when using within-country data in France (r=-0.27), Ireland (r=-0.23), the UK (r=-0.79) and Canada (r=-0.62). In the USA, the positive correlation (r=0.26) between a state's proportion of Catholics and its breast feeding initiation rate was confounded by race, education and socioeconomic status (SES). After controlling for education and SES, the state proportion of non-Hispanic white Catholics was negatively correlated (r=-0.29) with the rate of breast feeding initiation. In this ecological study, we found consistent negative correlations between Catholicism and breast feeding initiation rates. Qualitative and quantitative studies at the individual level are needed to confirm and explain our findings. Our results suggest that women living in a country or region where Catholicism has historically dominated are less likely to initiate breast feeding, and that breast feeding promotion policies should be adapted to

  14. A clinical study of integrating acupuncture and Western medicine in treating patients with Parkinson's disease.

    Science.gov (United States)

    Chen, Fang-Pey; Chang, Ching-Mao; Shiu, Jing-Huei; Chiu, Jen-Hwey; Wu, Ta-Peng; Yang, Jen-Lin; Kung, Yen-Ying; Chen, Fun-Jou; Chern, Chang-Ming; Hwang, Shinn-Jang

    2015-01-01

    Complementary therapy with acupuncture for Parkinson's disease (PD) has been studied for quite a long time, but the effectiveness of the treatment still remains unclear. The aim of this study is to evaluate the integrated effects of acupuncture treatment in PD patients who received western medicine. In the short-term acupuncture treatment study, 20 patients received acupuncture therapy twice a week in acupoints DU 20, GB 20, LI 11, LI 10, LI 4, GB 31, ST 32, GB 34 and GB 38 along with western medicine for 18 weeks, and 20 controlled patients received western medicine only. In the long-term acupuncture treatment, 13 patients received acupuncture treatment twice a week for 36 weeks. The outcome parameters include Unified Parkinson's disease rating scale (UPDRS), Beck Anxiety Inventory (BAI), Beck Depression Inventory-Version 2 (BDI-II), and WHO quality of life (WHOQOL). In the short-term clinical trial, a higher percentage of patients in the acupuncture group had score improvement in UPDRS total scores (55% vs. 15%, p = 0.019), sub-score of mind, behavior and mood (85% vs. 25%, p vs. 15%, p = 0.003), mobility (40% vs. 15%, p = 0.155) and complication of treatment (75% vs. 15%, p vs. 35%, p = 0.003), and WHOQOL score (65% vs. 15%, p = 0.003) when compared to control group at the end of the 18 weeks' follow up. After 36 weeks of long-term acupuncture treatment, the mean UPDRS total scores and sub-score of mentation, behavior and mood, sub-score of complications of therapy and BDI-II score decreased significantly when compared to the pretreatment baseline. In conclusion, acupuncture treatment had integrated effects in reducing symptoms and signs of mind, behavior, mood, complications of therapy and depression in PD patients who received Western medicine.

  15. Acute lead poisoning in western Canadian cattle - A 16-year retrospective study of diagnostic case records.

    Science.gov (United States)

    Cowan, Vanessa; Blakley, Barry

    2016-04-01

    This study describes the epidemiology of acute lead poisoning in western Canadian cattle over the 16-year period of 1998 to 2013 and reports background bovine tissue lead concentrations. Case records from Prairie Diagnostic Services, Western College of Veterinary Medicine, identified 525 cases of acute lead toxicity over the investigational period. Poisonings were influenced by year (P poisoned (53.5%; P poisoned. Mean toxic lead concentrations (mg/kg wet weight) in the blood, liver, and kidney were 1.30 ± 1.70 (n = 301), 33.5 ± 80.5 (n = 172), and 56.3 ± 39.7 (n = 61). Mean normal lead concentrations in the blood, liver, and kidney were 0.036 ± 0.003 mg/kg (n= 1081), 0.16 ± 0.63 mg/kg (n = 382), and 0.41 ± 0.62 mg/kg (n = 64).

  16. NEW PROBLEMATIZATION WAYS OF THE GREAT PATRIOTIC WAR HISTORY IN WESTERN RUSSIAN STUDIES

    OpenAIRE

    Наталья Валерьевна Трубникова

    2013-01-01

    The article examines the new thematic perspectives prevailing in Western historiography of the Great Patriotic War. The analysis is implemented on the basis of the French scientific periodicals of 2001–2013 years. Historical survey of contemporary Russian Studies seeking to fill research gaps and form a new problem fields in the study of the World War II Eastern Front history. These studies continue the trend of the second half of 1990 – the beginning of the 2000s, which withdrow themes of So...

  17. Sensitivity of the International Monitoring System infrasound network to elevated sources: a western Eurasia case study

    Science.gov (United States)

    Nippress, Alexandra; Green, David N.

    2017-11-01

    For the past 5 years (2010-2015) infrasound arrivals have been included in International Data Centre analyst-reviewed bulletins of events detected across the International Monitoring System (IMS). In western Eurasia, there are clusters of up to 268 events that consist of only infrasound arrivals (no associated seismic phases). These clusters are of unknown origin, although one in the North Sea region is associated with sonic booms from supersonic aircraft activity. IMS data for 17 North Sea events are analysed and compared with data from the Large Aperture Infrasound Array in the Netherlands to support the existence of these events and to determine common characteristics. Three other large clusters in western Eurasia are also identified and studied and show similar characteristics to the North Sea events, indicative of supersonic aircraft activity. The IMS infrasound network is shown to be particularly sensitive to sonic booms because the elevated source height reduces the anisotropy of infrasonic propagation within a stratospheric duct and allows for episodic upwind propagation. This episodic upwind propagation in addition to the prevailing downwind propagation, leads to clusters of Reviewed Event Bulletin events with constrained locations in western Eurasia region during the summer months. In the winter months, the recorded arrivals suggest that episodic upwind propagation is not as prevalent. Propagation modelling indicates that the subsequent unidirectional propagation, combined with the sparseness of the IMS network, leads to elongated lines of estimated event locations.

  18. Physical self-concept of adolescents in Western Balkan countries: a pilot study.

    Science.gov (United States)

    Janić, Snežana Radisavljević; Jurak, Gregor; Milanović, Ivana; Lazarević, Dušanka; Kovač, Marjeta; Novak, Dario

    2014-10-01

    The aim of this study was to explore physical self-concept of adolescents of the Western Balkans (Serbia, Slovenia, Croatia, and Bosnia and Herzegovina) according to sex and country. The participants were 2,606 students, ages 13 and 14 years (M = 13.5, SD = 0.9). The Physical Self-Description Questionnaire (PSDQ) was used to assess multidimensional physical self-concept. The results show the interaction of sex and country for three dimensions of physical self-concept (Appearance, Global Physical Self-Concept, and Self-Esteem). It was shown that female and male adolescents' perception of physical appearance, self-esteem, and global physical self-concept is more susceptible to influences of socio-cultural factors in the monitored countries. In all other dimensions of Physical self-concept, sex differences were consistently manifested in favour of male adolescents, except in Flexibility. Regardless of adolescents' sex, under the increasing influence of Western culture in the Western Balkan countries, adolescents more critically evaluate their body and motor abilities.

  19. Two-Dimensional Model Test Study of New Western Breakwater Proposal for Port of Hanstholm

    DEFF Research Database (Denmark)

    Eldrup, Mads Røge; Andersen, Thomas Lykke

    The present report presents results from a two-dimensional model test study carried out at Aalborg University in December 2016 with the proposed trunk section for the new western breakwater in Port of Hanstholm. The objectives of the model tests were to study the stability of the armour layer, to...... erosion, overtopping and transmission. The scale used for the model tests was 1:61.5. Unless otherwise specified all values given in this report are prototype values converted from the model to prototype according to the Froude model law....

  20. Physical Impacts of Climate Change on the Western US Electricity System: A Scoping Study

    Energy Technology Data Exchange (ETDEWEB)

    Coughlin, Katie; Goldman, Charles

    2008-12-01

    This paper presents an exploratory study of the possible physical impacts of climate change on the electric power system, and how these impacts could be incorporated into resource planning in the Western United States. While many aspects of climate change and energy have been discussed in the literature, there has not yet been a systematic review of the relationship between specific physical effects and the quantitative analyses that are commonly used in planning studies. The core of the problem is to understand how the electric system is vulnerable to physical weather risk, and how to make use of information from climate models to characterize the way these risks may evolve over time, including a treatment of uncertainty. In this paper, to provide the necessary technical background in climate science, we present an overview of the basic physics of climate and explain some of the methodologies used in climate modeling studies, particularly the importance of emissions scenarios. We also provide a brief survey of recent climate-related studies relevant to electric system planning in the Western US. To define the institutional context, we discuss the core elements of the resource and reliability planning processes used currently by utilities and by the Western Electricity Coordinating Council. To illustrate more precisely how climate-related risk could be incorporated into modeling exercises, we discuss three idealized examples. Overall, we argue that existing methods of analysis can and should be extended to encompass the uncertainties related to future climate. While the focus here is on risk related to physical impacts, the same principles apply to a consideration of how future climate change policy decisions might impact the design and functioning of the electric grid. We conclude with some suggestions and recommendations on how to begin developing this approach within the existing electric system planning framework for the West.

  1. Western Wind and Solar Integration Study Phase 3A: Low Levels of Synchronous Generation

    Energy Technology Data Exchange (ETDEWEB)

    Miller, Nicholas W. [GE Energy Management, Atlanta, GA (United States); Leonardi, Bruno [GE Energy Management, Atlanta, GA (United States); D' Aquila, Robert [GE Energy Management, Atlanta, GA (United States); Clark, Kara [National Renewable Energy Lab. (NREL), Golden, CO (United States)

    2015-11-17

    The stability of the North American electric power grids under conditions of high penetrations of wind and solar is a significant concern and possible impediment to reaching renewable energy goals. The 33% wind and solar annual energy penetration considered in this study results in substantial changes to the characteristics of the bulk power system. This includes different power flow patterns, different commitment and dispatch of existing synchronous generation, and different dynamic behavior from wind and solar generation. The Western Wind and Solar Integration Study (WWSIS), sponsored by the U.S. Department of Energy, is one of the largest regional solar and wind integration studies to date. In multiple phases, it has explored different aspects of the question: Can we integrate large amounts of wind and solar energy into the electric power system of the West? The work reported here focused on the impact of low levels of synchronous generation on the transient stability performance in one part of the region in which wind generation has displaced synchronous thermal generation under highly stressed, weak system conditions. It is essentially an extension of WWSIS-3. Transient stability, the ability of the power system to maintain synchronism among all elements following disturbances, is a major constraint on operations in many grids, including the western U.S. and Texas systems. These constraints primarily concern the performance of the large-scale bulk power system. But grid-wide stability concerns with high penetrations of wind and solar are still not thoroughly understood. This work focuses on 'traditional' fundamental frequency stability issues, such as maintaining synchronism, frequency, and voltage. The objectives of this study are to better understand the implications of low levels of synchronous generation and a weak grid on overall system performance by: 1) Investigating the Western Interconnection under conditions of both high renewable

  2. Gastrointestinal lymphoma in Western Algeria: pattern of distribution and histological subtypes (retrospective study).

    Science.gov (United States)

    Zeggai, Soumia; Harir, Noria; Tou, Abdenacer; Medjamia, Miloud; Guenaoui, Khaira

    2016-12-01

    Primary gastrointestinal (GI) lymphomas (GIL) are uncommon diseases that can involve the whole GI tract. Considerable variation exists in the literature with respect to incidence of the various histological subtypes and sites of involvement. This study was undertaken to establish the anatomic distribution, histological subtypes and sites of GI lymphomas of patients from Western Algeria. The case records of 58 consecutive patients with GIL diagnosed at the Pathologies Departments of Algerian west region (the Military Hospital of Oran city and the Central University Hospital of Sidi Bel Abbes city) from January 2006 to December 2013 were retrospectively evaluated for epidemiology and histopathology report. All lymphomas were reclassified according to the WHO 2008 classification. A total of 58 patients (39 male, 19 female) with mean age of 61 years and a range of 20-89 years were included in this study. Stomach was the most common site involved (70.7%). The commonest histological subtype was mucosa-associated lymphoid tissue (MALT) B cell lymphoma (46.6%), followed by diffuse large B-cell lymphomas (DLBCL) (43.1%).The frequency of Helicobacter pylori (HP) positivity differ between gastric and intestinal location P=0.003 and correlates with the histological type P=0.01. This retrospective study of patients with GI lymphoma from Western Algeria illustrates the pattern of distribution of various common and rare histological subtypes. More studies are necessary to find a potential cause, risk factor or genetic mutation that can explain these specific characteristics of GIL.

  3. INTANGIBLE CAPITAL, INNOVATION AND EXPORT-LED GROWTH: EMPIRICAL COMPARATIVE STUDY OF SLOVENIA AND THE WESTERN BALKANS

    National Research Council Canada - National Science Library

    Marija Drenkovska; Tjasa Redek

    2015-01-01

    .... In a comparative study we investigate the intellectual capital of a sample of firms from the Western Balkans and Slovenia, and analyse the link between intellectual capital, innovation, and export volume...

  4. Study of movement of the western and central belts of Peninsular Malaysia using GPS data analysis

    Energy Technology Data Exchange (ETDEWEB)

    Ramli, Siti Hafizah; Samsudin, Abdul Rahim [Geology Programme, School of Environmental and Natural Resource Sciences, Faculty of Science and Technology, Universiti Kebangsaan Malaysia, 43600 Bangi, Selangor (Malaysia)

    2014-09-03

    Since the large earthquakes in Sumatera and Nias, there were some tremors incidents at Bukit Tinggi. Therefore, a study on the earth’s crust movement and the effects of the earthquake in Indonesia on the tectonic blocks of Peninsular Malaysia have been carried out using GPS data analysis. GPS data from five MyRTKnet stations within Peninsular Malaysia have been analyzed to monitor the movement of two major tectonic blocks of Peninsular Malaysia which are the western belt represented by the Behrang (BEHR) and UPM Serdang (UPMS) stations and the central belt represented by Bentong (BENT), Jerantut (JRNT) and Temerloh (TLOH) stations. GPS data recorded from 2005 to 2010 were analysed based on horizontal and vertical displacements of the respective stations by using Trimble Business Centre (TBC) software. Based on the results of accumulated displacements of recorded GPS data from January 2006 to December 2013, it shows that the western belt which represented by UPMS has shifted 0.096m towards northwest with changes of ellipsoidal height of +0.030m while the central belt which represented by TLOH has shifted 0.080m towards northwest with changes of ellipsoidal height of −0.015m. Meanwhile, BENT station which is located on the Bentong-Raub suture zone turns to its original position as well as JRNT station. However, BEHR station which are located in western belt do not show any movements. All of these movements may be due to the influence of reactive faults in the stations area stimulated by several large earthquakes that occurred in 2005 to 2010. Study on using the GPS data analysis and combine with integrated geophysical methods are necessary to understand in detail about the tectonic evolution of Peninsular Malaysia.

  5. A long term (1999-2008) study of radar anomalous propagation conditions in the Western Mediterranean

    Science.gov (United States)

    Magaldi, A. V.; Mateu, M.; Bech, J.; Lorente, J.

    2016-03-01

    In this paper a study of the radio propagation environment of electromagnetic waves prevailing in the lower troposphere of the Western Mediterranean basin is presented. Deviations from atmospheric average or standard radio propagation conditions (anomalous propagation or AP) can affect significantly the quality of weather radar observations and other telecommunication systems. This is particularly important when ducting or superrefraction is present and spurious echoes resulting from the interaction of the beam with the ground or sea surface may appear. These AP conditions occur mainly when temperature inversions or strong moisture gradients are present. The period covered in this study ranges from 1999 to 2008 and conditions were derived from the Weather Research and Forecasting (WRF) modeling system, using the Japanese 25-year Reanalysis (JRA25) dataset as initial and boundary data. From the WRF model, we use the temperature, moisture, and pressure fields with a grid length of 30-km horizontal resolution and 250 m vertical resolution to compute several indices such as the Vertical Refractivity Gradient, Vertical Modified Refractivity Gradient and a Ducting Index. Results obtained show that on the Western Mediterranean coast the most favorable conditions for superrefraction are found in summer, while the most affected areas are the Gulf of Valencia, the Strait of Gibraltar and the Northern Gulf of Lion. Additionally, a comparison with radiosonde data recorded in Barcelona (NE Spain) is also performed indicating an overall agreement between model and observational data despite a tendency to decrease subrefractive events by the WRF model.

  6. Study on gastrointestinal zoonotic parasites in pet dogs in Western Iran.

    Science.gov (United States)

    Gharekhani, Jamal

    2014-01-01

    Dogs are the definitive or reservoirs hosts of more than 60 zoonotic parasites. This study was conducted to investigate the prevalence of gastrointestinal (GI) parasites in pet dogs in Hamedan, Western Iran. In cross-sectional study, 210 stool samples were collected randomly in pet dogs without clinical signs in Hamedan in April to December 2010. All samples were concentrated by formalin-ether technique. Smears of the feces were prepared and stained with Ziehl-Neelsen, trichrome, and iodine stains. During coproscopy, the overall proportion of GI parasitic infection was found in 6.7% (14/210) of samples. The detected parasites with their frequencies were Cryptosporidium spp. (3.8%), Toxocara canis (1.9%), and Giardia spp. (0.95%). A significant difference was not observed between infection rates in different age groups) p=0.617) or between genders (p=0.627). This is the first report of GI parasites in dogs from Western Iran. Although the rate of infection is low, the results showed that the pet dogs are reservoirs for zoonotic GI parasites and should be considered important to public health in this region. A combination of routine screening fecal samples for parasites, strategic anthelmintics regimens, and improved pet owner education is highly recommended for the control of GI parasites in pet dogs.

  7. A modeling study of climate variability over western and eastern Africa

    Science.gov (United States)

    Sun, Liqiang

    1997-09-01

    This dissertation is composed of six separate papers linked by a common purpose: to investigate the physical mechanisms associated with the climate variability over western and eastern Africa. This study is carried out on both monthly and seasonal time-scales. In Chapter 2, the NASA GEOS-1 GCM was adopted to investigate the role of orography in determining the western African climate. The moist southwesterlies emanating from the Atlas-Ahaggar orographic trough and the dry hot northeasterlies originating from the corresponding orographic windward ridge tend to converge thus re-enforcing the ITCZ over the Sahel. A zonal orographic ridge is generated along the coastal region of West Africa. Thus a permanent orographic induced rainfall dipole pattern over western Africa is produced. A weaker orographic induced rainfall dipole pattern across West Africa is simulated in response to the 1973 SST anomaly pattern. Consequently, wetter conditions along the coastal region and rainfall deficits over the Sahel are produced, which are consistent with observations. In Chapter 3, we customized the NCAR Regional Climate Model (RegCM2) for eastern Africa. The physics of RegCM2 has been improved. In Chapter 4, the model simulated both large-scale and mesoscale features during the autumn rains of 1988. In Chapter 5, we simulated the interannual variability of precipitation during autumn rains between 1982 and 1993. The stronger Arabian High appears to be responsible for earlier onset of rains, and the enhanced St. Helena and weaker Mascarene Highs lead to positive precipitation anomalies over Tanzania. The enhanced St. Helena High also leads to positive precipitation anomalies over Angola plateau. Strong connection between precipitation anomalies over Lake Victoria and ENSO exists. El Nino events usually lead to positive precipitation anomalies over the Turkana Channel. Precipitation over western Kenya Highlands is significantly affected by Lake Victoria and the Arabian High. The

  8. Three-month follow-up of Western and non-Western participants in a study on preconceptional ancestry-based carrier couple screening for cystic fibrosis and hemoglobinopathies in the Netherlands.

    NARCIS (Netherlands)

    Lakeman, P.; Plass, A.M.C.; Henneman, L.; Bezemer, P.D.; Cornel, M.C.; Kate, L.P. ten

    2008-01-01

    Objective: To study psychological outcomes, knowledge, recall and understanding of test-results, satisfaction, and reproductive intentions among 97 Western and 46 non-Western participants in a unique preconceptional carrier screening study for both cystic fibrosis and hemoglobinopathies in a

  9. Three-month follow-up of Western and non-Western participants in a study on preconceptional ancestry-based carrier couple screening for cystic fibrosis and hemoglobinopathies in the Netherlands

    NARCIS (Netherlands)

    Lakeman, P.; Plass, A.M.C.; Henneman, L.; Bezemer, P.D.; Cornel, M.C.; ten Kate, L.P.

    2008-01-01

    Objective: To study psychological outcomes, knowledge, recall and understanding of test-results, satisfaction, and reproductive intentions among 97 Western and 46 non-Western participants in a unique preconceptional carrier screening study for both cystic fibrosis and hemoglobinopathies in a

  10. Process Philosophy and the Text-Image Interface: A Study of Three Western Australian Botanical Illustrators

    Directory of Open Access Journals (Sweden)

    John Charles Ryan

    2012-12-01

    Full Text Available Botanical illustration combines scientific knowledge and artistic technique. However, whereas illustrated botanical images record static visual qualities, such as form and color, written botanical narratives supply crucial sensory, ecological, historical, and cultural contexts that complement visual representation. Understanding the text-image interface—where images and words intersect—contributes to humanities-based analyses of botanical illustration and illustrators. More specifically, a process philosophy perspective reveals the extent to which botanical representations engage the temporality, cyclicality, and contextuality of the living plants being illustrated. This article takes up these themes through a comparative theoretical study of three female Western Australian botanical illustrators, Georgiana Leake (1812–1869, Emily Pelloe (1877–1941, and Philippa Nikulinsky (born 1942, whose lives together span the 183 year history of the Swan River Colony and the state of Western Australia. I apply a processist framework to examine the text-image interface of their works. All three illustrators use some form of textuality: marginalia, annotations, written accompaniments, introductory statements, and other narrative materials. In examining their written commentaries and traces, I identify the emergence of a process mode of botanical illustration that represents plants as ecological, historical, cultural, and temporal organisms.

  11. Developing High PV Penetration Cases for Frequency Response Study of U.S. Western Interconnection: Preprint

    Energy Technology Data Exchange (ETDEWEB)

    Tan, Jin; Zhang, Yingchen; Veda, Santosh; Elgindy, Tarek; Liu, Yilu

    2017-04-11

    Recent large penetrations of solar photovoltaic (PV) generation and the inertial characteristics of inverter-based generation technologies have caught the attention of those in the electric power industry in the United States. This paper presents a systematic approach to developing test cases of high penetrations of PV for the Western Interconnection. First, to examine the accuracy of the base case model, the Western Electricity Coordinating Council (WECC) model is validated by using measurement data from synchronized phasor measurement units. Based on the 2022 Light Spring case, we developed four high PV penetration cases for the WECC system that are of interest to the industry: 5% PV+15 % wind, 25% PV+15% wind, 45% PV+15% wind, 65% PV+15% wind). Additionally, a method to project PV is proposed that is based on collected, realistic PV distribution information, including the current and future PV power plant locations and penetrations in the WECC system. Both the utility-scale PV plant and residential rooftop PV are included in this study.

  12. The Western Wind and Solar Integration Study Phase 2 (Executive Summary)

    Energy Technology Data Exchange (ETDEWEB)

    Lew, Debra [National Renewable Energy Lab. (NREL), Golden, CO (United States); Brinkman, Greg [National Renewable Energy Lab. (NREL), Golden, CO (United States)

    2013-09-01

    The electric grid is a highly complex, interconnected machine, and changing one part of the grid can have consequences elsewhere. Adding wind and solar affects the operation of the other power plants and adding high penetrations can induce cycling of fossil-fueled generators. Cycling leads to wear-and-tear costs and changes in emissions. Phase 2 of the Western Wind and Solar Integration Study (WWSIS-2) evaluated these costs and emissions and simulated grid operations for a year to investigate the detailed impact of wind and solar on the fossil-fueled fleet. This built on Phase 1, one of the largest wind and solar integration studies ever conducted, which examined operational impacts of high wind and solar penetrations in the West(GE Energy 2010).

  13. Typological analysis of social linear blocks: Spain 1950-1983. The case study of western Andalusia

    Directory of Open Access Journals (Sweden)

    A. Guajardo

    2017-04-01

    Full Text Available A main challenge that cities will need to face in the next few years is the regeneration of the social housing estates built during the decades of 1950s, 1960s and 1970s. One of the causes of their obsolescence is the mismatch between their hous-ing typologies and the contemporary needs. The main target of this study is to contribute to take a step forward in the un-derstanding of these typologies to be able to intervene on them efficiently. With this purpose, a study on 42 linear blocks built in Spain between 1950 and 1983 in western Andalusia has been carried out. The analysis includes three stages: 1 classification of the houses in recognizable groups; 2 an identification of the most used spatial configurations and 3 definition of their programmatic and size characteristics. As a result, a characterization of linear blocks is proposed as a reference model for future regenerative interventions.

  14. Streptavidin-Binding Peptide (SBP-tagged SMC2 allows single-step affinity fluorescence, blotting or purification of the condensin complex

    Directory of Open Access Journals (Sweden)

    Graham Alison N

    2010-12-01

    Full Text Available Abstract Background Cell biologists face the need to rapidly analyse their proteins of interest in order to gain insight into their function. Often protein purification, cellular localisation and Western blot analyses can be multi-step processes, where protein is lost, activity is destroyed or effective antibodies have not yet been generated. Aim To develop a method that simplifies the critical protein analytical steps of the laboratory researcher, leading to easy, efficient and rapid protein purification, cellular localisation and quantification. Results We have tagged the SMC2 subunit of the condensin complex with the Streptavidin-Binding Peptide (SBP, optimising and demonstrating the efficacious use of this tag for performing these protein analytical steps. Based on silver staining, and Western analysis, SBP delivered an outstanding specificity and purity of the condensin complex. We also developed a rapid and highly specific procedure to localise SBP-tagged proteins in cells in a single step procedure thus bypassing the need for using antibodies. Furthermore we have shown that the SBP tag can be used for isolating tagged proteins from chemically cross-linked cell populations for capturing DNA-protein interactions. Conclusions The small 38-amino acid synthetic SBP offers the potential to successfully perform all four critical analytical procedures as a single step and should have a general utility for the study of many proteins and protein complexes.

  15. Amphibian responses to wildfire in the western united states: Emerging patterns from short-term studies

    Science.gov (United States)

    Hossack, B.R.; Pilliod, D.S.

    2011-01-01

    The increased frequency and severity of large wildfires in the western United States is an important ecological and management issue with direct relevance to amphibian conservation. Although the knowledge of fire effects on amphibians in the region is still limited relative to most other vertebrate species, we reviewed the current literature to determine if there are evident patterns that might be informative for conservation or management strategies. Of the seven studies that compared pre- and post-wildfire data on a variety of metrics, ranging from amphibian occupancy to body condition, two reported positive responses and five detected negative responses by at least one species. Another seven studies used a retrospective approach to compare effects of wildfire on populations: two studies reported positive effects, three reported negative effects from wildfire, and two reported no effects. All four studies that included plethodontid salamanders reported negative effects on populations or individuals; these effects were greater in forests where fire had been suppressed and in areas that burned with high severity. Species that breed in streams are also vulnerable to post-wildfire changes in habitat, especially in the Southwest. Wildfire is also important for maintaining suitable habitat for diverse amphibian communities, although those results may not be evident immediately after an area burns. We expect that wildfire will extirpate few healthy amphibian populations, but it is still unclear how populations will respond to wildfire in the context of land management (including pre- and post-fire timber harvest) and fragmentation. Wildfire may also increase the risk of decline or extirpation for small, isolated, or stressed (e.g., from drought or disease) populations. Improved understanding of how these effects vary according to changes in fire frequency and severity are critical to form more effective conservation strategies for amphibians in the western United States.

  16. No difference in stroke knowledge between Korean adherents to traditional and western medicine – the AGE study: an epidemiological study

    Directory of Open Access Journals (Sweden)

    Eun Su-Yong

    2006-06-01

    Full Text Available Abstract Background Effective stroke intervention and risk reduction depend on the general public's awareness and knowledge of stroke. In Korea, where both traditional Oriental medicine and Western medicine are practiced, estimates of the general public's awareness and knowledge of stroke are poor. The present study sought to describe the inception cohort of the Ansan Geriatric Study (AGE study and to determine baseline stroke awareness and preferred medical treatment for stroke in this Korean sample. Methods A total of 2,767 subjects selected randomly from the Ansan Geriatric Study in South Korea were questioned about stroke. Their answers were compared with their sociodemographic data and other variables. Results Only 44.8% of participants correctly identified stroke as a vascular disease in the human brain. Sudden numbness or weakness was the most frequently identified stroke warning sign (60.2%. Hypertension (66.7% and mental stress (62.2% were most frequently identified as stroke risk factors. The contributions of diabetes mellitus and cardiovascular disease to stroke were underestimated; they were identified as risk factors by 28.3% and 18.6% of participants, respectively. The predictors for poor knowledge of stroke warning signs and risk factors were similar irrespective of preference for Western or Oriental medical treatment, and included those with lower levels of education and inaccurate definition of stroke. Television and radio (40.3% were the most frequent sources of stroke information for both groups. Conclusion This study shows that knowledge of stroke is similar among Koreans with preferences for either Western or Oriental medical treatment and that misunderstandings about stroke are common among the Korean elderly. In order to prevent and manage stroke effectively, public health education regarding basic concepts of stroke is necessary. This should target those with a lower level of education and a misunderstanding of the

  17. A case study of butterfly road kills from Anaikatty Hills, Western Ghats, Tamil Nadu, India

    Directory of Open Access Journals (Sweden)

    R. K. Sony

    2015-11-01

    Full Text Available  Anaikatty Hills of the Western Ghats in Tamil Nadu witness the annual spectacle of mass movement of lakhs of butterflies.  The present paper examines the impact of vehicular traffic on this ‘butterfly migration’ through a survey of butterfly mortality along a road stretch in Anaikatty Hills.  A high rate of mortality due to road traffic was observed during the mass movement of butterflies.  One-hundred-and-thirty-five butterfly road kills belonging to three families, nine genera and 12 species were recorded during the study.  The proportion of nymphalid butterflies among the road kills (70% was very high compared to their respective share in the background population (39%, indicating a higher road mortality risk for nymphalids.  The conservation significance of the road traffic impact on butterfly assemblage and management options are discussed. 

  18. 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.

  19. Western Wind and Solar Integration Study Phase 3 – Frequency Response and Transient Stability

    Energy Technology Data Exchange (ETDEWEB)

    Miller, N. W. [GE Energy Management, Schenectady, NY (United States); Shao, M. [GE Energy Management, Schenectady, NY (United States); Pajic, S. [GE Energy Management, Schenectady, NY (United States); D' Aquila, R. [GE Energy Management, Schenectady, NY (United States)

    2014-12-01

    Power system operators and utilities worldwide have concerns about the impact of high-penetration wind and solar generation on electric grid reliability (EirGrid 2011b, Hydro-Quebec 2006, ERCOT 2010). The stability of North American grids under these conditions is a particular concern and possible impediment to reaching future renewable energy goals. Phase 3 of the Western Wind and Solar Integration Study (WWSIS-3) considers a 33% wind and solar annual energy penetration level that results in substantial changes to the characteristics of the bulk power system, including different power flow patterns, different commitment and dispatch of existing synchronous generation, and different dynamic behavior of wind and solar generation. WWSIS-3 evaluates two specific aspects of fundamental frequency system stability: frequency response and transient stability.

  20. Landscape ecological planning through a multi-scale characterization of pattern: studies in the Western Ghats, South India.

    Science.gov (United States)

    Nagendra, Harini; Utkarsh, Ghate

    2003-09-01

    This article analyzes landscape pattern in the Western Ghats mountain ranges in southwestern India at two scales, comparing small-scale, detailed studies of landscape pattern, with broader, regional-scale assessments of the Western Ghats. Due in large part to their inaccessibility, relatively little is known about the landscapes of this biodiverse region, which also supports some of the highest population densities in the world. A broad-scale NDVI-based IRS 1B satellite image classification is used to analyze north-south and east-west trends across the entire Western Ghats and western coast of India, an area over 170000 km2. Northern and eastern landscapes are more fragmented compared to the southern and western slopes. Western slopes also have greater landscape diversity with land cover types more interspersed compared to the eastern slopes. These differences can be related to north-south and east-west variations in rainfall and plant distribution. Data from thirteen landscapes 10-50 km2 in area, are further utilized to analyze trends in landscape pattern, and describe the geographical distribution of major natural and managed ecotope types. At this scale, very high levels of intra-ecotope type variability in landscape pattern are encountered for all land cover types. Results at these two scales are integrated to suggest a hierarchical stratified approach for monitoring land cover and biodiversity in the region.

  1. From Western TESOL Classrooms to Home Practice: A Case Study with Two "Privileged" Saudi Teachers

    Science.gov (United States)

    Barnawi, Osman Z.; Phan, Le Ha

    2015-01-01

    This article is located in the debates concerning the continued problems underlying the cultural politics of English-speaking Western countries' Teaching English to Speakers of Other Languages (TESOL) programmes and "Western" pedagogies. It examines two Saudi TESOL teachers' pedagogical enactments in their home teaching contexts after…

  2. Emergency care facility access in rural areas within the golden hour?: Western Cape case study.

    Science.gov (United States)

    Vanderschuren, Marianne; McKune, Duncan

    2015-01-16

    Road Safety is a major cause of death around the world and South Africa has one of the highest road fatality rates. Many measures, engineering and medical, are investigated. However, analysis of the accessibility of emergency care facilities is often overlooked. This paper aims to fill the gap between pre-crash engineering solutions and literature on trauma injuries and emergency care procedures. The focus is on the role that accessibility to emergency care facilities in rural areas plays, given that 50% of the world's population lives in rural areas, which are often omitted from international research. The Western Cape (a rural province with low population volumes and high volume roads in South Africa) is analysed as an example of access to trauma care in rural areas. It is internationally accepted that the time to emergency care facilities influences the survival chances. However, the international literature still debates the exact time period. In this paper, the 'Golden Hour' is used to analyse the accessibility of emergency care facilities in rural areas and establish a geographical analysis method which identifies risk areas. The analysis can be repeated if the international literature debates regarding the exact time period changes. A Geographical Information System (GIS) tool revealed that 53% of the fatalities in the rural parts of the Western Cape occur outside the Golden Hour. In high risk crash areas, the fatality risk is up to nine times higher than the province's rural average. People in need of trauma care after a road crash are most likely to survive if they receive definitive care timeously. At the time of the study, the rural areas in the Western Cape had 44 Emergency Medical Services stations and 29 medical facilities that can assist to provide definitive (trauma) care. Further optimisation of the facility locations is recommended and research has begun.More advanced geographical modelling is possible when improved data becomes available on the

  3. [Comparative study on Chinese medicine and western medicine for treatment of prolapse of lumbar intervertebral disc].

    Science.gov (United States)

    Qin, Xin-yan; Li, Xiao-xia; Suteanu, Stefan

    2007-05-01

    To compare therapeutic effects, safety and tolerance of TCM, western medicine and integrated Chinese and western medicine for treatment of acute lumbosacral pain induced by prolapse of lumbar intervertebral disc. Ninety cases were randomly divided into 3 groups, 30 cases in each group. They were treated respectively with western medicine, TCM and combined TCM and western medicine, and the pain intensity, activity, muscular tension, and other indexes were monitored after 7 days and 30 days of treatment. After treatment of 7 days, the combined treatment group in improvement of VAS scores of lumbosacral pain and radiating pain of the lower limbs was superior to the TCM group with no significant difference between the two groups, and in improvement of VAS scores of lumbosacral pain and radiating pain of the lower limbs, Lasegue's sign, activity of spinal column (Schober test and distance from finger tip to floor), etc. were superior to the western medicine group (P prolapse of lumbar intervertebral disc.

  4. Development of improved wildfire smoke exposure estimates for health studies in the western U.S.

    Science.gov (United States)

    Ivey, C.; Holmes, H.; Loria Salazar, S. M.; Pierce, A.; Liu, C.

    2016-12-01

    Wildfire smoke exposure is a significant health concern in the western U.S. because large wildfires have increased in size and frequency over the past four years due to drought conditions. The transport phenomena in complex terrain and timing of the wildfire emissions make the smoke plumes difficult to simulate using conventional air quality models. Monitoring data can be used to estimate exposure metrics, but in rural areas the monitoring networks are too sparse to calculate wildfire exposure metrics for the entire population in a region. Satellite retrievals provide global, spatiotemporal air quality information and are used to track pollution plumes, estimate human exposures, model emissions, and determine sources (i.e., natural versus anthropogenic) in regulatory applications. Particulate matter (PM) exposures can be estimated using columnar aerosol optical depth (AOD), where satellite AOD retrievals serve as a spatial surrogate to simulate surface PM gradients. These exposure models have been successfully used in health effects studies in the eastern U.S. where complex mountainous terrain and surface reflectance do not limit AOD retrival from satellites. Using results from a chemical transport model (CTM) is another effective method to determine spatial gradients of pollutants. However, the CTM does not adequately capture the temporal and spatial distribution of wildfire smoke plumes. By combining the spatiotemporal pollutant fields from both satellite retrievals and CTM results with ground based pollutant observations the spatial wildfire smoke exposure model can be improved. This work will address the challenge of understanding the spatiotemporal distributions of pollutant concentrations to model human exposures of wildfire smoke in regions with complex terrain, where meteorological conditions as well as emission sources significantly influence the spatial distribution of pollutants. The focus will be on developing models to enhance exposure estimates of

  5. Natural Airborne Dust and Heavy Metals: A Case Study for Kermanshah, Western Iran (2005–2011)

    Science.gov (United States)

    PIRSAHEB, Meghdad; ZINATIZADEH, Aliakbar; KHOSRAVI, Touba; ATAFAR, Zahra; DEZFULINEZHAD, Saeed

    2014-01-01

    Abstract Background Dust pollution has become a serious environmental problem especially in recent decades. The present study aim was the investigation of the levels of PM10 concentration in Kermanshah, western Iran and also measured five important heavy metals (Pb, Cd, As, Hg and Cr) in some samples during 2005 to 2011. Methods A total 2277 samples were collected from air pollution measurement station belonging to the Department of Environment in Kermanshah. Furthermore, four samples were collected during dusty days to determine the selected heavy metals concentration. The samples were analyzed statistically using the SPSS Ver.16 Results The highest seasonal average concentration in spring was recorded in 2008 with 216.63μg/m3, and the maximum values of 267.79 and 249.09μg/m3 were observed in summer and winter in 2009, respectively. The maximum concentration of 127.1μg/m3 was in autumn in 2010. The metals concentration (Pb, Cd, As, Hg and Cr) of samples were 42.32±5.40, 37.45±9.29, 3.51±2.07, 1.88±1.64 and 0μg/g in July, 2009, respectively. Conclusion According to National Ambient Air Quality of USEPA guidelines, the most days with non-standard, warning, emergency and critical conditions were related to 2009 (120 days) while the least polluted days were recorded in 2006 (16 days). There are concerns about the increasing frequency and intensity trend of dust storms in recent years as a result of special condition in neighboring Western countries which it could endanger public health and environment. All measured heavy metals except mercury was higher than the standard level of WHO and USEPA. PMID:26005656

  6. Natural airborne dust and heavy metals: a case study for kermanshah, Western iran (2005-2011).

    Science.gov (United States)

    Pirsaheb, Meghdad; Zinatizadeh, Aliakbar; Khosravi, Touba; Atafar, Zahra; Dezfulinezhad, Saeed

    2014-04-01

    Dust pollution has become a serious environmental problem especially in recent decades. The present study aim was the investigation of the levels of PM10 concentration in Kermanshah, western Iran and also measured five important heavy metals (Pb, Cd, As, Hg and Cr) in some samples during 2005 to 2011. A total 2277 samples were collected from air pollution measurement station belonging to the Department of Environment in Kermanshah. Furthermore, four samples were collected during dusty days to determine the selected heavy metals concentration. The samples were analyzed statistically using the SPSS Ver.16. The highest seasonal average concentration in spring was recorded in 2008 with 216.63μg/m(3), and the maximum values of 267.79 and 249.09μg/m(3) were observed in summer and winter in 2009, respectively. The maximum concentration of 127.1μg/m(3) was in autumn in 2010. The metals concentration (Pb, Cd, As, Hg and Cr) of samples were 42.32±5.40, 37.45±9.29, 3.51±2.07, 1.88±1.64 and 0μg/g in July, 2009, respectively. According to National Ambient Air Quality of USEPA guidelines, the most days with non-standard, warning, emergency and critical conditions were related to 2009 (120 days) while the least polluted days were recorded in 2006 (16 days). There are concerns about the increasing frequency and intensity trend of dust storms in recent years as a result of special condition in neighboring Western countries which it could endanger public health and environment. All measured heavy metals except mercury was higher than the standard level of WHO and USEPA.

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

    Energy Technology Data Exchange (ETDEWEB)

    Mansfield, E.S.; Worley, J.M. [Molecular Dynamics, Inc., Sunnyvale, CA (United States); Zimmerman, P.A. [Laboratory of Parasitic Diseases, Bethesda, MD (United States)] [and others

    1994-09-01

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

  8. A Field Study of Plague and Tularemia in Rodents, Western Iran.

    Science.gov (United States)

    Mostafavi, Ehsan; Shahraki, Abdolrazagh Hashemi; Japoni-Nejad, Alireza; Esmaeili, Saber; Darvish, Jamshid; Sedaghat, Mohammad Mehdi; Mohammadi, Ali; Mohammadi, Zeinolabedin; Mahmoudi, Ahmad; Pourhossein, Behzad; Ghasemi, Ahmad; Gyuranecz, Miklós; Carniel, Elisabeth

    2017-04-01

    Kurdistan Province in Iran is a historical focus for plague and tularemia. This study aimed at assessing the current status of these two foci by studying their rodent reservoirs. Rodents were trapped and their ectoparasites were collected. The genus and species of both rodents and ectoparasites were determined. Serological analyses of rodent blood samples were done by enzyme-linked immunosorbent assay for plague and by standard tube agglutination assay for tularemia. Rodent spleen samples were subjected to bacterial culture, microscopic examination, and real-time PCR to search for active plague or tularemia infection. During this study, 245 rodents were trapped, of which the most abundant genera were Apodemus (40%), Mus (24.49%), and Meriones (12.65%). One hundred fifty-three fleas, 37 mites, and 54 ticks were collected on these rodents. The results of all direct and indirect tests were negative for plague. Serological tests were positive for tularemia in 4.8% of trapped rodents. This study is the first report on the presence of tularemia infection in rodents in Western Iran. Since Meriones persicus is a known reservoir for plague and tularemia, and this rodent carried plague and tularemia vectors in Marivan and Sanandaj districts, there is a real potential for the occurrence of these two diseases in this region.

  9. Influence of Prescribed Herbal and Western Medicine on Patients with Abnormal Liver Function Tests: A Retrospective Quasi-Experimental Study

    Science.gov (United States)

    Lee, Ah-Ram; Yim, Je-Min; Kim, Won-Il

    2012-01-01

    Objectives: The aim of this study was to investigate the safety and the efficacy of Korean herbal, western and combination medicine use in patients with abnormal liver function tests. Methods: We investigated nerve disease patients with abnormal liver function tests who were treated with Korean herbal, western and combination medicine at Dong-Eui University Oriental Hospital from January 2011 to August 2011. We compared aspartic aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and total bilirubin (T-bil) levels before and after taking medicine and excluded patients who had liver-related disease when admitted. Results: AST and ALT were decreased significantly in patients who had taken herbal, western medicine. AST, ALT and ALP were decreased significantly in patients who had taken combination medicine. Compare to herbal medicine, AST, ALT and ALP were decreased significantly in patients who had taken western medicine, and ALT and ALP were decreased significantly in patients who had taken combination medicine. There were no significant differences between western and combination medicine. Conclusions: This study suggests that prescribed Korean herbal medicine, at least, does not injure liver function for patients’, moreover, it was shown to be effective in patients with abnormal liver function tests. PMID:25780634

  10. Influence of prescribed herbal and Western medicine on patients with abnormal liver function tests: a retrospective quasi-experimental study.

    Science.gov (United States)

    Lee, Ah-Ram; Yim, Je-Min; Kim, Won-Il

    2012-03-01

    The aim of this study was to investigate the safety and the efficacy of Korean herbal, western and combination medicine use in patients with abnormal liver function tests. We investigated nerve disease patients with abnormal liver function tests who were treated with Korean herbal, western and combination medicine at Dong-Eui University Oriental Hospital from January 2011 to August 2011. We compared aspartic aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP) and total bilirubin (T-bil) levels before and after taking medicine and excluded patients who had liver-related disease when admitted. AST and ALT were decreased significantly in patients who had taken herbal, western medicine. AST, ALT and ALP were decreased significantly in patients who had taken combination medicine. Compare to herbal medicine, AST, ALT and ALP were decreased significantly in patients who had taken western medicine, and ALT and ALP were decreased significantly in patients who had taken combination medicine. There were no significant differences between western and combination medicine. This study suggests that prescribed Korean herbal medicine, at least, does not injure liver function for patients', moreover, it was shown to be effective in patients with abnormal liver function tests.

  11. Influence of Prescribed Herbal and Western Medicine on Patients with Abnormal Liver Function Tests: A Retrospective Quasi-Experimental Study

    Directory of Open Access Journals (Sweden)

    Lee Ah-Ram

    2012-03-01

    Full Text Available Objectives: The aim of this study was to investigate the safety and the efficacy of Korean herbal, western and combination medicine use in patients with abnormal liver function tests. Methods: We investigated nerve disease patients with abnor-mal liver function tests who were treated with Korean herbal, western and combination medicine at Dong-Eui University Oriental Hospital from January 2011 to August 2011. We co-mpared aspartic aminotransferase (AST, alanine aminotran-sferase (ALT, alkaline phosphatase (ALP and total bilirubin (T-bil levels before and after taking medicine and excluded patients who had liver-related disease when admitted. Results: AST and ALT were decreased significantly in patients who had taken herbal, western medicine. AST, ALT and ALP were decreased significantly in patients who had taken comb-ination medicine. Compare to herbal medicine, AST, ALT and ALP were decreased significantly in patients who had taken western medicine, and ALT and ALP were decreased signifi-cantly in patients who had taken combination medicine. There were no significant differences between western and combin-ation medicine. Conclusions: This study suggests that prescribed Korean herbal medicine, at least, does not injure liver function for patients’, moreover, it was shown to be effective in patients with abnormal liver function tests.

  12. Study of Endemic and Threatened Fish Species Diversity and its Assemblage Structure from Northern Western Ghats, Maharashtra, India

    OpenAIRE

    Tejas S. Patil; Amrut R. Bhosale; Rupesh B. Yadav; Rupali S. Khandekar; Dipak V. Muley

    2015-01-01

    The present investigation undertakes to study endemic and threatened fish species and its assemblage structure from rivers of Kolhapur district for the period July-2012 to December-2014. The study area is situated in the extreme Southern part of Maharashtra state. It contributes much more part of Western Ghats. A total of 23 species belonging to 7 families and 19 genera were reported. In which 9 species are threatened and 20 species are endemic to Western Ghats, we found that 6 species are th...

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

    Energy Technology Data Exchange (ETDEWEB)

    Lee, C.Y.G. (Univ. of British Columbia, Vancouver, Canada); Huang, Y.S.; Hu, P.C.; Gomel, V.; Menge, A.C.

    1982-06-01

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

  14. Examining Culture's Impact on the Learning Behaviors of International Students from Confucius Culture Studying in Western Online Learning Context

    Science.gov (United States)

    Kang, Haijun; Chang, Bo

    2016-01-01

    There is a lack of shared understanding of how culture impacts learning in online environment. Utilizing document analysis, the authors in this research study culture's impact on the learning behaviors of student sojourners from Confucius culture studying in Western online learning context. The shared understandings of Confucius culture and…

  15. Questioning western assessment of trauma among Tibetan torture survivors. A quantitative assessment study with comments from Buddhist Lamas

    DEFF Research Database (Denmark)

    Elsass, Peter; Carlsson, Jessica; Jespersen, Kristian

    2009-01-01

    Our study falls in line with the numerous studies providing a critique of the use of western diagnostic instruments for assessing trauma in a cross-cultural context. Our purpose has been to give evidence for the Tibetan torture survivors' degree of traumatisation and for their use of spirituality...

  16. The Evolutions of Interest and Beliefs about Arabic as a Foreign Language: A Case Study on Three Western Learners

    Science.gov (United States)

    Rashed, Hazem

    2013-01-01

    The purpose of this case study was to investigate the evolutions of interest and beliefs about Arabic as a foreign language among three Western faculty members at an education college in United Arab Emirates. The study used four data resources which were: interviews, field notes, reflective journals and an information form. After four months of…

  17. Tap into Good Teeth--a Western Australian pilot study of children's drinking patterns.

    Science.gov (United States)

    Leavy, Justine E; Heyworth, Jane; Middleton, Aves; Rosenberg, Michael; Woloszyn, Magdalene

    2012-04-01

    ISSUED ADDRESSED: The increase trend in the prevalence of dental caries in Australian children is a concern to public health professionals. Attitudes, behaviours and lifestyle patterns established in childhood are often carried throughout adult life. The objective of the study was to estimate the proportion of Perth metropolitan year two public primary school children drinking tap water at home, school and play. It also aimed to explore knowledge and attitudes that children and parents have towards drinking tap water, bottled water, fruit juices and soft drinks. Nine Western Australian government primary schools were recruited. A facilitator-led questionnaire was administered to year two primary school students and a matched parent self-administered questionnaire was also completed. Forty-two per cent of the children in our study reported if thirsty they drank tap (fluoridated) water at home whereas parents stated 60% of children drank tap water at home. The type of drink appeared to vary with time of day/activity while overall water was most frequently drunk; a higher proportion of milk was drunk at breakfast, whereas soft drinks were drunk in a greater proportion while watching television. This study found the vast majority of year two children in metropolitan Perth public primary schools are drinking tap water.

  18. US-China collaborative study of Western Tibet: progress report and initial results

    Science.gov (United States)

    Shokoohi Razi, A.; Huang, G.; Levin, V. L.; Roecker, S. W.; Ping, X.; Cao, Z.

    2009-12-01

    While Tibetan plateau has been the focus of numerous geophysical studies to date, logistics of access to the region resulted in the majority of them being concentrated on the eastern half of the plateau. At the same time global-scale studies suggest lateral variability in properties beneath the seemingly uniform plateau. We are conducting a passive seismological study to improve the degree of understanding of the western side of the Tibetan plateau, with crustal and uper mantle structure being the key objectives. Since the summer of 2007 we operate a broadband array in western Tibet, between longitudes of 79 and 83 deg. East. The array started with 10 nodes, and has been densified by additional 20 nodes in late summer of 2009. We expect the larger array to operate through the summer of 2011. We retrieved and archived data from the first 12 months of recording. Using these data, we develop preliminary constraints on the crustal structure of the region. We use P and S wave travel time data from local earthquakes to develop a 1D velocity model by simultaneously inverting for vertical velocity distribution and earthquake locations. Having examined records of over 300 events present in the Chinese national catalog, we retained data from 268 events that yielded approximately 1200 arrivals at nine sites for which data were recovered. A large fraction of these earthquakes are almost due north from our array, within the intense aftershock sequence that followed a March, 2008 M=7.2 earthquake on the TIbet -Tarim Basin border. Preliminary assessment of apparent P wave speed obtained using catalog earthquake locations showed a large scatter most likely caused by uncertainties in hypocentral parameters. We also use records of teleseismic P waves to develop crustal thickness constraints using receiver function analysis. Results we obtain are similar to other areas in TIbet, with a likely Moho-converted Ps phase seen at 8-10 s delay. A site near the Himalaya also shows evidence

  19. Ethics of Surrogacy: A Comparative Study of Western Secular and Islamic Bioethics

    Science.gov (United States)

    Islam, Sharmin; Nordin, Rusli Bin; Bin Shamsuddin, Ab Rani; Mohd Nor, Hanapi Bin; Al-Mahmood, Abu Kholdun

    2012-01-01

    The comparative approach regarding the ethics of surrogacy from the Western secular and Islamic bioethical view reveals both commensurable and incommensurable relationship. Both are eager to achieve the welfare of the mother, child and society as a whole but the approaches are not always the same. Islamic bioethics is straightforward in prohibiting surrogacy by highlighting the lineage problem and also other social chaos and anarchy. Western secular bioethics is relative and mostly follows a utilitarian approach. PMID:23864994

  20. Crosscultural differences in frontalis muscle tension levels: an exploratory study comparing Japanese and Westerners.

    Science.gov (United States)

    Uchiyama, K; Lutterjohann, M; Shah, M D

    1981-03-01

    This paper describes a comparison between Japanese (N = 20) and Westerners (N = 20) in their respective frontal EMG levels and their ability to relax during one 5-minute biofeedback training session. The data indicated a significantly lower baseline muscle tension level in Japanese than in Westerners. After EMG biofeedback, however, no significant difference was found between the two groups. Japanese females showed significantly higher tension levels both during baseline and biofeedback phases than did japanese males.

  1. Mortality in migrants living with HIV in western Europe (1997-2013): a collaborative cohort study.

    Science.gov (United States)

    2015-12-01

    Many migrants face adverse socioeconomic conditions and barriers to health services that can impair timely HIV diagnosis and access to life-saving treatments. We aimed to assess the differences in overall mortality by geographical origin in HIV-positive men and women using data from COHERE, a large European collaboration of HIV cohorts from 1997 to 2013. In this observational cohort study, we included HIV-positive, antiretroviral-naive people accessing care in western Europe from COHERE. Individuals were eligible if enrolled in a cohort that collected information on geographical origin or ethnic origin from Jan 1, 1997, to March 19, 2013, aged 18-75 years, they had available information about sex, they were not infected perinatally or after the receipt of clotting factor concentrates, and were naive to combination antiretroviral therapy at cohort entry. Migrants' origins were grouped into seven regions: western Europe and similar countries (Australia, Canada, New Zealand, and the USA); eastern Europe; North Africa and the Middle East; sub-Saharan Africa; Latin America; the Caribbean; and Asia and the rest of Oceania (excluding Australia and New Zealand). Crude and adjusted mortality rate ratios were calculated by use of Poisson regression stratified by sex, comparing each group with the native population. Multiple imputation with chained equations was used to account for missing values. Between Oct 25, 1979, and March 19, 2013, we recruited 279 659 individuals to the COHERE collaboration in EuroCoord. Of these 123 344 men and 45 877 women met the inclusion criteria. Our data suggested effect modification by transmission route (pinteraction=0·12 for men; pinteraction=0·002 for women). No significant difference in mortality was identified by geographical origin in men who have sex with men. In heterosexual populations, most migrant men had mortality lower than or equal to that of native men, whereas no group of migrant women had mortality lower than that in

  2. Floristic Study of Buxus hyrcana Stands in the Western Forests of Haraz District, Amol

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    S. Hosseinzadeh

    2017-06-01

    Full Text Available Floristic composition of boxwood (Buxus hyrcana Pojark stands in the forests of Western Haraz, distributed at the altitudinal range of 250 to 1,200 m asl, was investigated by field-walk method. Results showed that the flora of this area includes 50 families, 69 genera, and 78 plant species. Rosaceae, Orchidaceae, Dryopteridaceae, Lamiaceae and Aspleniaceae are the greatest families in this area. Chorological studies showed that the largest proportion of the flora is related to Euro-Siberian region (56.4%. Cryptophytes (32 species, Phanerophytes (26 sp. and Hemicryptophytes (16 sp., which are compromising 44%, 33.3% and 20.5% of the flora, were the most important structure groups of the biological spectrum. Abundance of Cryptophytes with Phanerophytes and Hemicryptophytes in these forests implys that there is a temperate climate with cold winter, frequently rainfall and relatively cool summer, which are suitable for growing temperate forest. Frequent occurrence of Prunus Laurocerasus trees with Daneae racemosa as a woody understory species especially in the upper parts of the studied area accompany by absence of Therophytes and relatively well distribution of ferns species, especially Asplenium scolopendrium, also implys high, favorable, moisture conditions without any disturbances in Box tree stands of these forests.

  3. Thrombocytopenia in malaria: its usefulness in a clinical setting. A study from Western Rajasthan, India.

    Science.gov (United States)

    Singh, Anurag; Goyal, Manish; Sharma, Deepak

    2014-10-01

    To assess the occurrence of thrombocytopaenia in hospitalised patients of malaria and to correlate its severity with the type of malaria in Western Rajasthan. A total of 100 patients with proven malaria were included in this study. Haematological parameters were determined by using an automated analyser. Fifty malaria-free patients were taken as controls. In the study group of 100 patients; 45 patients had P. vivax malaria, 38 had P. Falciparum malaria and mixed malaria was detected in 17 patients. Out of 100 patients, 89 patients had thrombocytopaenia while out of 50 controls only eight (16%) patients had thrombocytopaenia (P < 0.001). Ninety-one percent of patients with thrombocytopaenia also had anaemia. Thrombocytopaenia was more commonly associated with complicated malaria compared to uncomplicated malaria (P < 0.001). Any patient in a malaria endemic zone presenting with acute febrile illness and having thrombocytopaenia and anaemia should be alerted to the possibility of malarial infection. © The Author(s) 2014 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav.

  4. Sacred Groves: Myths, Beliefs, and Biodiversity Conservation—A Case Study from Western Himalaya, India

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    Sushma Singh

    2017-01-01

    Full Text Available Religious and traditional beliefs, cultural mores, and practices play a crucial role in the conservation of environment and biodiversity. The present paper describes a case study of two sacred groves in Western Himalaya. Sacred groves (SGs are patches of land that are communally protected with religious zeal. A preliminary survey was conducted in these SGs to study their role in biodiversity conservation. The data collected included the general information regarding the SGs and the associated deity, nearest human habitation, access to them, and their floral and faunal diversity. Ethnomedicinal property of plants was collected from the indigenous communities. Many taboos are associated with both the SGs, which help in managing resources well through ritual representation. Different festivals are organized, where the local communities reaffirm their commitment to the forest and the deity. Sacred groves, in general, are a valuable tool of biodiversity conservation. But people’s changing attitudes, erosion of traditional beliefs, and human impact have caused degradation of sacred groves over the years. Their conservation would not be possible without the active participation of the local people. By improving their living standards and by giving benefits of conservation to them, long-term conservation goals in these SGs can be achieved.

  5. Compilation of selected hydrologic data, through water year 1992, Black Hills Hydrology Study, western South Dakota

    Science.gov (United States)

    Driscoll, Daniel G.; Bradford, Wendell

    1994-01-01

    This report presents water-level, water-quailty, and springflow data that have been collected or compiled, through water year 1992, for the Black Hills Hydrology Study. This study is a long-term cooperative effort between the U.S. Geological Survey, the South Dakota Department of Environmental and Natural Resources, and the West Dakota Water Development District (which represents various local and county cooperators). Water-level data are presented for 32 observation wells and 2 cave sites in the Black Hills area of western South Dakota. The wells are part of a network of observation wells maintained by the South Dakota Department of Environment and Natural Resources and are completed in various bedrock formations that are utilized as aquifers in the Black Hills area. Both cave sites are located within outcrops of the Madison Limestone. Data presented include site descriptions, hydrographs, and tabular data. Water- quality data are presented for 12 surface-water sites and 5 ground-water sites. Data presented include field parameters, bacteria counts, and concentrations of common ions, solids, nutrients, trace elements, radiometrics, cyanide, phenols, dissolved organic carbon, and suspended sediment. Spring data are presented for 83 springs and 21 stream reaches with significant springflow components. Data presented include site information, discharge, and field water-quality parameters including temperature, specific conductance, dissolved oxygen, and pH.

  6. Early life programming of cardiometabolic disease in the Western Australian pregnancy cohort (Raine) study.

    Science.gov (United States)

    Huang, Rae-Chi; Mori, Trevor A; Beilin, Lawrence J

    2012-11-01

    The Raine study (http://www.rainestudy.org.au, accessed 18 June 2012) is a longitudinal Australian birth cohort that has serially assessed the offspring of 2900 pregnant women from 18 weeks gestation in utero to 17 years of age. The Raine study data have shown that low birth weight is a surrogate for poor in utero growth from 18 weeks gestation. A U-shaped relationship between birth size and cardiometabolic risk exists in this Western Australian cohort, implying that both low and high birth weight are associated with increased risk. High birth weight is a risk factor for cardiometabolic risk, particularly for females. Lifetime adiposity trajectories are better at predicting metabolic risk of the offspring than birth size or current body mass index. Therefore, early life programming is an ongoing process, starting in utero and undergoing at least some level of modification in parallel with changes in adiposity during early childhood. Maternal smoking during pregnancy, maternal obesity, hypertension and diabetes increase the risk for metabolic risk in the offspring. Breast feeding is protective for cardiometabolic risk in this Australian cohort. © 2012 The Authors Clinical and Experimental Pharmacology and Physiology © 2012 Wiley Publishing Asia Pty Ltd.

  7. Environmental Collaborations Between Indigenous Communities and Western Science: Case Studies and Reflections

    Science.gov (United States)

    Emanuel, R. E.

    2016-12-01

    The study of coupled natural and human systems in a changing world can benefit greatly from indigenous perspectives, which have the potential to bring deep, placed-based understanding to complex environmental issues while promoting sustainable solutions to pressing socio-environmental problems. In recent years, scientists have begun to embrace indigenous knowledge and perspectives, but indigenous voices in the sciences remain relatively few. At the same time, indigenous communities face wide ranging and unique vulnerabilities to global environmental change on a variety of fronts, particularly where water resources are concerned. Given this situation, indigenous scientists often find themselves bridging both western scientific and indigenous communities, sometimes embodying the nexus in a literal sense. Here I reflect on this nexus from the perspective of an indigenous hydrologist collaborating with American Indian communities in North Carolina, which has the largest American Indian population of any state in the eastern US. Intertwining case studies of coupled natural and human systems illustrate some of the the challenges, complexities, and successes of ongoing collaborations with tribal communities and Native-serving organizations on water resource issues, environmental impacts of food and energy production, and broadening participation of American Indians in the sciences.

  8. Economic Assessment of Overtopping BReakwater for Energy Conversion (OBREC: A Case Study in Western Australia

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    Pasquale Contestabile

    2016-12-01

    Full Text Available This paper constructs an optimal configuration assessment, in terms of the financial returns, of the Overtopping BReakwater for wave Energy Conversion (OBREC. This technology represents a hybrid wave energy harvester, totally embedded in traditional rubble mound breakwaters. Nine case studies along the southern coast of Western Australia have been analysed. The technique provides tips on how to estimate the quality of the investments, for benchmarking with different turbine strategy layouts and overlapping with the costs of traditional rubble mound breakwaters. Analyses of the offshore and nearshore wave climate have been studied by a high resolution coastal propagation model, forced with wave data from the European Centre for Medium-Range Weather Forecasts (ECMWF. Inshore wave conditions have been used to quantify the exploitable resources. It has been demonstrated that the optimal investment strategy is nonlinearly dependent on potential electricity production due to outer technical constraints. The work emphasizes the importance of integrating energy production predictions in an economic decision framework for prioritizing adaptation investments.

  9. Magnetotelluric Study of the Southern Margin of the Capricorn Orogen, Western Australia

    Science.gov (United States)

    Piña-Varas, P.; Dentith, M. C.

    2015-12-01

    We present the current status and the preliminary results of a magnetotelluric (MT) study that is being performed in the Capricorn Orogen, in northern Western Australia. This project is a component of large multi-disciplinary geoscience project on the 'Distal Footprints of Giant Ore Systems'. The Capricorn Orogen, a largely Proterozoic orogenic belt, is being used as a natural laboratory to develop methods for exploring for mineral deposits 'under cover'. The MT dataset consists of a total of 240 broadband magnetotelluric stations (BBMT) and 84 long period stations (LMT). Analysis of the dataset reveals the area to have complex 3-D variations in electrical conductivity, making 3-D interpretation methods essential. The first 3-D inversion model encompasses south-eastern part of the Capricorn Orogen (Padbury-Bryah Basins) and the adjoining Yilgarn Craton, and involves the inversion of 58 BBMT sites. The results shown a prominent resistive feature that is tentatively correlated with the northern boundary of the Yilgarn Craton. Apart from this, the other main geoelectrical feature is a high conductivity body located within the Yilgarn Craton. This structure could be related to tectonic features associated with the accretion of the different terranes composing this part of the craton. Forward models were used to validate these and other structures. Major faults occurring at cratonic margins are known to be important indicators of increased prospectivity and our preliminary results suggest MT surveys are a useful tool for regional scale exploration in the study area.

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

    Science.gov (United States)

    Volpi, Nicola; Maccari, Francesca

    2009-01-01

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

  11. Malaria mosquito control using edible fish in western Kenya: preliminary findings of a controlled study

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    Omlin Francois X

    2007-08-01

    Full Text Available Abstract Background Biological control methods are once again being given much research focus for malaria vector control. This is largely due to the emerging threat of strong resistance to pesticides. Larvivorous fish have been used for over 100 years in mosquito control and many species have proved effective. In the western Kenyan highlands the larvivorous fish Oreochromis niloticus L. (Perciformes: Cichlidae (formerly Tilapia nilotica is commonly farmed and eaten but has not been previously tested in the field for malaria mosquito control. Methods This fish was introduced into abandoned fishponds at an altitude of 1,880 m and the effect measured over six months on the numbers of mosquito immatures. For comparison an untreated control pond was used. During this time, all ponds were regularly cleared of emergent vegetation and fish re-stocking was not needed. Significant autocorrelation was removed from the time series data, and t-tests were used to investigate within a pond and within a mosquito type any differences before and after the introduction of O. niloticus. Mulla's formula was also used on the raw data to calculate the percentage reduction of the mosquito larvae. Results After O. niloticus introduction, mosquito densities immediately dropped in the treated ponds but increased in the control pond. This increase was apparently due to climatic factors. Mulla's formula was applied which corrects for that natural tendency to increase. The results showed that after 15 weeks the fish caused a more than 94% reduction in both Anopheles gambiae s.l. and Anopheles funestus (Diptera: Culicidae in the treated ponds, and more than 75% reduction in culicine mosquitoes. There was a highly significantly reduction in A. gambiae s.l. numbers when compared to pre-treatment levels. Conclusion This study reports the first field trial data on O. niloticus for malaria mosquito control and shows that this species, already a popular food fish in western Kenya

  12. Burn injury, gender and cancer risk: population-based cohort study using data from Scotland and Western Australia.

    Science.gov (United States)

    Duke, Janine M; Bauer, Jacqui; Fear, Mark W; Rea, Suzanne; Wood, Fiona M; Boyd, James

    2014-01-17

    To investigate the risk of cancer and potential gender effects in persons hospitalised with burn injury. Population-based retrospective cohort study using record-linkage systems in Scotland and Western Australia. Records of 37 890 and 23 450 persons admitted with a burn injury in Scotland and Western Australia, respectively, from 1983 to 2008. Deidentified extraction of all linked hospital morbidity records, mortality and cancer records were provided by the Information Service Division Scotland and the Western Australian Data Linkage Service. Total and gender-specific number of observed and expected cases of total ('all sites') and site-specific cancers and standardised incidence ratios (SIRs). From 1983 to 2008, for female burn survivors, there was a greater number of observed versus expected notifications of total cancer with 1011 (SIR, 95% CI 1.3, 1.2 to 1.4) and 244 (SIR, 95% CI 1.12, 1.05 to 1.30), respectively, for Scotland and Western Australia. No statistically significant difference in total cancer risk was found for males. Significant excesses in observed cancers among burn survivors (combined gender) in Scotland and Western Australian were found for buccal cavity, liver, larynx and respiratory tract and for cancers of the female genital tract. Results from the Scotland data confirmed the increased risk of total ('all sites') cancer previously observed among female burn survivors in Western Australia. The gender dimorphism observed in this study may be related to the role of gender in the immune response to burn injury. More research is required to understand the underlying mechanism(s) that may link burn injury with an increased risk of some cancers.

  13. Communication ability in cerebral palsy: a study from the CP register of western Sweden.

    Science.gov (United States)

    Himmelmann, Kate; Lindh, Karin; Hidecker, Mary Jo Cooley

    2013-11-01

    Communication is often impaired in cerebral palsy (CP). Tools are needed to describe this complex function, in order to provide effective support. To study communication ability and the relationship between the Communication Function Classification System (CFCS) and CP subtype, gross motor function, manual ability, cognitive function and neuroimaging findings in the CP register of western Sweden. Sixty-eight children (29 girls), 14 with unilateral spastic CP, 35 with bilateral spastic CP and 19 with dyskinetic CP, participated. The CFCS, Gross Motor Function Classification System (GMFCS) and Manual Ability Classification System (MACS) levels, cognitive impairment and neuroimaging findings were recorded. Half the children used speech, 32% used communication boards/books and 16% relied on body movements, eye gaze and sounds. Twenty-eight per cent were at the most functional CFCS level I, 13% at level II, 21% at level III, 10% at level IV and 28% at level V. CFCS levels I-II were found in 71% of children with unilateral spastic CP, 46% in bilateral spastic CP and 11% in dyskinetic CP (p = 0.03). CFCS correlated with the GMFCS, MACS and cognitive function (p CP can be derived from the CFCS, which correlates to gross and fine motor and cognitive function. Good communication ability is associated with lesions acquired early, rather than late, in the third trimester. Copyright © 2013 European Paediatric Neurology Society. Published by Elsevier Ltd. All rights reserved.

  14. An investigation of the auditory perception of western lowland gorillas in an enrichment study.

    Science.gov (United States)

    Brooker, Jake S

    2016-09-01

    Previous research has highlighted the varied effects of auditory enrichment on different captive animals. This study investigated how manipulating musical components can influence the behavior of a group of captive western lowland gorillas (Gorilla gorilla gorilla) at Bristol Zoo. The gorillas were observed during exposure to classical music, rock-and-roll music, and rainforest sounds. The two music conditions were modified to create five further conditions: unmanipulated, decreased pitch, increased pitch, decreased tempo, and increased tempo. We compared the prevalence of activity, anxiety, and social behaviors between the standard conditions. We also compared the prevalence of each of these behaviors across the manipulated conditions of each type of music independently and collectively. Control observations with no sound exposure were regularly scheduled between the observations of the 12 auditory conditions. The results suggest that naturalistic rainforest sounds had no influence on the anxiety of captive gorillas, contrary to past research. The tempo of music appears to be significantly associated with activity levels among this group, and social behavior may be affected by pitch. Low tempo music also may be effective at reducing anxiety behavior in captive gorillas. Regulated auditory enrichment may provide effective means of calming gorillas, or for facilitating active behavior. Zoo Biol. 35:398-408, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  15. Vessel traffic safety in busy waterways: A case study of accidents in western shenzhen port.

    Science.gov (United States)

    Mou, J M; Chen, P F; He, Y X; Yip, Tsz Leung; Li, W H; Tang, J; Zhang, H Z

    2016-08-03

    Throughout the world, busy waterways near large ports witness heavy vessel traffic in recent decades. The waterways are characterized by high risk in terms of loss of life, property, and pollution to environment. To facilitate maritime safety management with satisfactory efficiency and efficacy, the authors propose a framework of safety indexes to evaluate the risk level in busy waterways according to the accident severity, fatality rate and special indicators of maritime transportation. The safety indexes consist of Safety Evaluation Index (SEI) and Safety Warning Index (SWI), and are derived from the proposed risk criteria of Chinese vessel traffic. As a case study, data on vessel traffic accidents reported in the Western Shenzhen Port, South China from 1995 to 2015 are analyzed. The actual risk level of this area during the period is calculated under the framework. The implementation of the safety indexes indicate that the risk criteria and safety indexes are practicable and effective for the vessel traffic management. The methodology based on long-term accident data can significantly support the risk analysis in the macroscopic perspective for busy ports and waterways, such that SWI can act as threshold to trigger actions, while SEI can act as an indicator to measure safety status. Copyright © 2016 Elsevier Ltd. All rights reserved.

  16. 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.

  17. The unit of study and the interpretation of ethnicity : studying the Nkoya of Western Zambia

    NARCIS (Netherlands)

    Binsbergen, W.M.J.

    1981-01-01

    The alternative proposed here for the tribal model as a unit of study is not another, better unit of study (e.g. a mode of production, an expanding social formation, or a well-defined spatio-temporal portion of reality), but a growing awareness of possible problems and interrelations, informed by

  18. 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.

  19. Mass anti-malarial administration in western Cambodia: a qualitative study of factors affecting coverage.

    Science.gov (United States)

    Pell, Christopher; Tripura, Rupam; Nguon, Chea; Cheah, Phaikyeong; Davoeung, Chan; Heng, Chhouen; Dara, Lim; Sareth, Ma; Dondorp, Arjen; von Seidlein, Lorenz; Peto, Thomas J

    2017-05-19

    Mass anti-malarial administration has been proposed as a key component of the Plasmodium falciparum malaria elimination strategy in the Greater Mekong sub-Region. Its effectiveness depends on high levels of coverage in the target population. This article explores the factors that influenced mass anti-malarial administration coverage within a clinical trial in Battambang Province, western Cambodia. Qualitative data were collected through semi-structured interviews and focus group discussions with villagers, in-depth interviews with study staff, trial drop-outs and refusers, and observations in the communities. Interviews were audio-recorded, transcribed and translated from Khmer to English for qualitative content analysis using QSR NVivo. Malaria was an important health concern and villagers reported a demand for malaria treatment. This was in spite of a fall in incidence over the previous decade and a lack of familiarity with asymptomatic malaria. Participants generally understood the overall study aim and were familiar with study activities. Comprehension of the study rationale was however limited. After the first mass anti-malarial administration, seasonal health complaints that participants attributed to the anti-malarial as "side effects" contributed to a decrease of coverage in round two. Staff therefore adapted the community engagement approach, bringing to prominence local leaders in village meetings. This contributed to a subsequent increase in coverage. Future mass anti-malarial administration must consider seasonal disease patterns and the importance of local leaders taking prominent roles in community engagement. Further research is needed to investigate coverage in scenarios that more closely resemble implementation i.e. without participation incentives, blood sampling and free healthcare.

  20. Suicidal ideation among students of a medical college in Western Nepal: a cross-sectional study.

    Science.gov (United States)

    Menezes, Ritesh G; Subba, S H; Sathian, Brijesh; Kharoshah, Magdy A; Senthilkumaran, Subramanian; Pant, Sadip; Arun, M; Kundapur, Rashmi; Jain, Animesh; Lobo, Stany Wilfred; Ravi Shankar, P

    2012-07-01

    Many studies have been conducted in the developed countries to know the magnitude and factors influencing suicidal ideation among medical students, but such data are sparse in developing countries. This cross-sectional study was therefore conducted to find out the prevalence of suicidal ideation and factors influencing such ideation among students of a medical college in Western Nepal. A total of 206 students were selected using random sampling and questioned about their socio-demographic factors, other risk factors and suicidal ideation using a preformed validated questionnaire. The data were analyzed using SPSS for Windows Version 16.0 and the EPI Info 3.5.1 Windows Version. Descriptive statistics and testing of hypothesis were applied for the statistical methodology. The univariate and multivariate logistic regression methods were used to examine the association between different variables. Suicidal ideation in the last one year was present in nearly one tenth of the study population and in almost one fifth of them life-time suicidal ideation was present. Factors that were associated with suicidal ideation were primarily dissatisfaction with academic performance, being in the clinical semesters, having history of drug abuse and feeling neglected by parents. Most common reason reported for suicidal ideation was family related followed by self-related. Recognition of suicidal ideation among students and their associated factors can help in detecting it on time, making the right interventions and controlling the problem. Understanding the magnitude of the problem and their epidemiology via scientific study like this would be the first step in this process. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  1. Variable density management in riparian reserves: lessons learned from an operational study in managed forests of western Oregon, USA.

    Science.gov (United States)

    Samuel Chan; Paul Anderson; John Cissel; Larry Lateen; Charley. Thompson

    2004-01-01

    A large-scale operational study has been undertaken to investigate variable density management in conjunction with riparian buffers as a means to accelerate development of late-seral habitat, facilitate rare species management, and maintain riparian functions in 40-70 year-old headwater forests in western Oregon, USA. Upland variable retention treatments include...

  2. A Study on the Functions of Western Cultural Non-Verbal Behavior in English Classroom in China

    Science.gov (United States)

    Wei, Yuehong

    2013-01-01

    In China, English classroom is the main place of English language acquisition. Therefore, how to improve English classroom teaching effectively has become the scholars' concern. This paper reports a study conducted at North China Electric Power University on the functions of western cultural nonverbal behaviors in English classroom in China.…

  3. Western conceptualizations and Eastern experience: A cross-cultural study of traumatic stress reactions among Tibetan refugees in India

    NARCIS (Netherlands)

    Terheggen, M.A.; Stroebe, M.S.; Kleber, R.J.

    This study investigated the nature and impact of traumatic experiences among Tibetan refugees in India. It explored the applicability of western conceptualizations of reactions to traumatic events among this cultural group. A randomly selected sample of refugee camp students was assessed on

  4. Western Conceptualizations and Eastern Experience : A Cross-cultural Study of Traumatic Stress Reactions among Tibetan Refugees in India

    NARCIS (Netherlands)

    Terheggen, M.A.; Stroebe, M.S.; Kleber, R.J.

    2001-01-01

    This study investigated the nature and impact of traumatic experiences among Tibetan refugees in India. It explored the applicability of western conceptualizations of reactions to traumatic events among this cultural group. A randomly selected sample of refugee camp students was assessed on measures

  5. Density management and riparian buffer study in Western Oregon: Phase 1 results, launch of phase 2 [brochure

    Science.gov (United States)

    Rhonda Mazza

    2009-01-01

    Can we expedite the development of late-successional forest conditions by applying thinning treatments to young forest stands? What effect will these thinning treatments have on headwater ecosystems? These broad questions lie at the foundation of the Density Management and Riparian Buffer Study (DMS) of western Oregon.

  6. Identifying Knowledge Sharing Barriers in the Collaboration of Traditional and Western Medicine Professionals in Chinese Hospitals: A Case Study

    Science.gov (United States)

    Zhou, Lihong; Nunes, Miguel Baptista

    2012-01-01

    This paper reports on a research project that aims at identifying knowledge sharing (KS) barriers between traditional and western medicine practitioners co-existing and complementing each other in Chinese healthcare organisations. The study focuses on the tacit aspects of patient knowledge, rather than the traditional technical information shared…

  7. Concurrent Study of Eastern and Western Medicine at the National College of Natural Medicine: Dual or Duel?

    Science.gov (United States)

    Smith, Andrea Christine

    2010-01-01

    Students at the National College of Natural Medicine (NCNM) are eligible to concurrently study both Western medicine, as reflected by the Doctor of Naturopathic Medicine (ND) program, and Eastern medicine, as exhibited by the Master of Science in Oriental Medicine (MSOM) degree program. The dual track is unique in that the dominant Western…

  8. [Comparative study on irritable bowel syndrome treated with acupuncture and western medicine].

    Science.gov (United States)

    Shi, Zhi-Min; Zhu, Ye-Shan; Wang, Qing-Xian; Lei, Miao-Na

    2011-07-01

    To compare the differences in the therapeutic effect on irritable bowel syndrome (IBS) between acupuncture at Tianshu (ST 25) and Dachangshu (BL 25) and western medication with Trimebutine Maleate. Forty cases were divided randomly into an acupuncture group and a western medication group, 20 cases in each one. In acupuncture group, acupuncture was applied to Tianshu (ST 25) and Dachangshu (BL 25). Ziwu Daojiu needling technique was adopted, once daily. In western medication group, Trimebutine Maleate capsule was administered, 2 capsules in each time, 3 times per day. The assessment on the therapeutic effect was performed in 4 weeks of treatment in two groups. As compared with those before treatment, the time of abdominal pain, the frequency of abdominal pain, the morbidity of abnormal stool appearance, the morbidity of defecation abnormality, the morbidity of mucus stool and the score of bloating or abdominal pain on bowel movement were all reduced after treatment in two groups (all P acupuncture group were much more significant than those in western medication group (the total score: 16.70 +/- 2.40 vs 15.70 +/- 3.01, P acupuncture group was 95.0% (19/20), which was superior to that of 70.0% (14/20) in western medication group (P Acupuncture at Tianshu (ST 25) and Dachangshu (BL 25) may remarkably relieve the clinical symptoms of IBS and its efficacy is superior to that of oral medication with Trimebutine Maleate.

  9. Western Australian Public Opinions of a Minimum Pricing Policy for Alcohol: Study Protocol.

    Science.gov (United States)

    Keatley, David A; Carragher, Natacha; Chikritzhs, Tanya; Daube, Mike; Hardcastle, Sarah J; Hagger, Martin S

    2015-11-18

    Excessive alcohol consumption has significant adverse economic, social, and health outcomes. Recent estimates suggest that the annual economic costs of alcohol in Australia are up to AUD $36 billion. Policies influencing price have been demonstrated to be very effective in reducing alcohol consumption and alcohol-related harms. Interest in minimum pricing has gained traction in recent years. However, there has been little research investigating the level of support for the public interest case of minimum pricing in Australia. This article describes protocol for a study exploring Western Australian (WA) public knowledge, understanding, and reaction to a proposed minimum price policy per standard drink. The study will employ a qualitative methodological design. Participants will be recruited from a wide variety of backgrounds, including ethnic minorities, blue and white collar workers, unemployed, students, and elderly/retired populations to participate in focus groups. Focus group participants will be asked about their knowledge of, and initial reactions to, the proposed policy and encouraged to discuss how such a proposal may affect their own alcohol use and alcohol consumption at the population level. Participants will also be asked to discuss potential avenues for increasing acceptability of the policy. The focus groups will adopt a semi-structured, open-ended approach guided by a question schedule. The schedule will be based on feedback from pilot samples, previous research, and a steering group comprising experts in alcohol policy and pricing. The study is expected to take approximately 14 months to complete. The findings will be of considerable interest and relevance to government officials, policy makers, researchers, advocacy groups, alcohol retail and licensed establishments and organizations, city and town planners, police, and other stakeholder organizations.

  10. Western Australian Public Opinions of a Minimum Pricing Policy for Alcohol: Study Protocol

    Science.gov (United States)

    Keatley, David A; Daube, Mike; Hardcastle, Sarah J

    2015-01-01

    Background Excessive alcohol consumption has significant adverse economic, social, and health outcomes. Recent estimates suggest that the annual economic costs of alcohol in Australia are up to AUD $36 billion. Policies influencing price have been demonstrated to be very effective in reducing alcohol consumption and alcohol-related harms. Interest in minimum pricing has gained traction in recent years. However, there has been little research investigating the level of support for the public interest case of minimum pricing in Australia. Objective This article describes protocol for a study exploring Western Australian (WA) public knowledge, understanding, and reaction to a proposed minimum price policy per standard drink. Methods The study will employ a qualitative methodological design. Participants will be recruited from a wide variety of backgrounds, including ethnic minorities, blue and white collar workers, unemployed, students, and elderly/retired populations to participate in focus groups. Focus group participants will be asked about their knowledge of, and initial reactions to, the proposed policy and encouraged to discuss how such a proposal may affect their own alcohol use and alcohol consumption at the population level. Participants will also be asked to discuss potential avenues for increasing acceptability of the policy. The focus groups will adopt a semi-structured, open-ended approach guided by a question schedule. The schedule will be based on feedback from pilot samples, previous research, and a steering group comprising experts in alcohol policy and pricing. Results The study is expected to take approximately 14 months to complete. Conclusions The findings will be of considerable interest and relevance to government officials, policy makers, researchers, advocacy groups, alcohol retail and licensed establishments and organizations, city and town planners, police, and other stakeholder organizations. PMID:26582408

  11. Development of a monoclonal antibody-based colony blot immunoassay for detection of thermotolerant Campylobacter species.

    Science.gov (United States)

    Huang, Hongsheng; Phipps-Todd, Beverley; McMahon, Tanis; Elmgren, Catherine L; Lutze-Wallace, Cheryl; Todd, Zoe A; Garcia, Manuel M

    2016-11-01

    Campylobacter species, particularly thermotolerant Campylobacter spp., such as C. jejuni, are major human foodborne pathogens. Culture methods have been routinely used for the detection of this organism in various types of samples. An alternative, simple and rapid confirmation test(s) without further tedious biochemical tests would be useful. Meanwhile, Campylobacter-like colonies can be difficult to identify on agar plates overgrown with competitive bacteria, which can lead to false-negative results. This study was to develop a simple colony blot immunoassay using a new monoclonal antibody (Mab) produced in the present study for rapid screening, confirmation and quantification of campylobacters on culture agar plates. The procedure developed in this study was able to specifically detect thermotolerant Campylobacter spp., but not other non-thermotolerant Campylobacter and non-Campylobacter reference strains tested. This assay could detect 105 cells in a single dot. This assay showed 100% correlation with the culture method for the blotted membranes from 21 either chicken meat or vegetable samples experimentally inoculated with thermotolerant campylobacters. Among 101 natural samples of chicken meat (n=44), chicken feces (n=20) and vegetables (n=37), this assay also showed positive for 23 chicken meat and 14 fecal samples that were positive for thermotolerant campylobacters by culture method, and identified four additional suspects that were culture negative. Membranes stored at 4°C for at least 4years could also be used for this assay. The assay developed in this study can be used in quantitative study for immediate or archival usage, and for diagnostic test to preliminarily confirm the presence of thermotolerant Campylobacter on agar plates. Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.

  12. Unaccompanied and Separated Foreign Children in the Care System in the Western Cape – A Socio-Legal Study

    Directory of Open Access Journals (Sweden)

    Julia Sloth-Nielsen

    2016-05-01

    Full Text Available This article reports on the findings of a study of foreign children accommodated in the care system in the Western Cape, based on fieldwork conducted in child and youth care centres. The objectives of the study were firstly to map and quantify the number and demographics of foreign children placed in all CYCCs across the Western Cape. Secondly, the study aimed to analyse the reasons for children's migration and the circumstances around their placement in residential care institutions in order to establish whether family reunification was possible or desirable. Thirdly, the study explores the sufficiency of efforts made to trace and reunify the children with their families, whether in South Africa or across borders, as the institutional placement of children should not only be a last resort but it should preferably be temporary whilst family-based solutions are sought. Lastly, the documentation status of the children in the study was examined. Recommendations emanating from the research conclude the study.

  13. Endophthalmitis in the western Sydney region: a case-control study.

    Science.gov (United States)

    Lertsumitkul, S; Myers, P C; O'Rourke, M T; Chandra, J

    2001-12-01

    A retrospective case-control study was conducted to investigate risk factors for endophthalmitis following routine intraocular surgery. A review was performed of consecutive cases of endophthalmitis from three teaching hospitals in the western Sydney region and matched controls from the same institutions between 1996 and 1998. There were 31 cases and 66 controls. Eighty procedures were phacoemulsification, 15 conventional extracapsular cataract extraction, and two were penetrating keratoplasties. Of the 80 patients who had phacoemulsification surgery, 50 had a clear corneal incision, and 26 had a scleral incision (four were unknown). Logistic regression showed an increased risk of endophthalmitis with surgical complications (P = 0.002) and clear cornea temporal incisions (P = 0.007). Risk of endophthalmitis was reduced with use of subconjunctival injections (P = 0.008). The yield for the Gram stain was 47% and for culture was 67%. Anterior chamber tap in addition to vitreous biopsy alone did not increase the yield for microorganism (P = 0.78). Mean visual acuity on presentation was hand movement with 13 patients (50%) showing visual improvement following intravitreal injections of antibiotics (P = 0.003). Visual prognosis did not correlate with presenting visual acuity but appeared to be better in those who grew Staphylococcus epidermidis or were culture negative. Although this study is unable to draw definite conclusions regarding risk of endophthalmitis in clear corneal temporal cataract surgery, sufficient data suggest the importance of incision type and location. Surgical complication is an important risk factor for endophthalmitis. Use of subconjunctival antibiotic injections at the conclusion of the procedure is recommended.

  14. Wintertime particulate pollution episodes in an urban valley of the Western US: a case study

    Directory of Open Access Journals (Sweden)

    L.-W. A. Chen

    2012-11-01

    Full Text Available This study investigates the causes of elevated PM2.5 episodes and potential exceedences of the US National Ambient Air Quality Standards (NAAQS in Truckee Meadows, Nevada, an urban valley of the Western US, during winter 2009/2010, an unusually cold and snowy winter. Continuous PM2.5 mass and time-integrated chemical speciation data were acquired from a central valley monitoring site, along with meteorological measurements from nearby sites. All nine days with PM2.5 > 35 μg m−3 showed 24-h average temperature inversion of 1.5–4.5 °C and snow cover of 8–18 cm. Stagnant atmospheric conditions limited wind ventilation while highly reflective snow cover reduced daytime surface heating creating persistent inversion. Elevated ammonium nitrate (NH4NO3 and water associated with it are found to be main reasons for the PM2.5 exceedances. An effective-variance chemical mass balance (EV-CMB receptor model using locally-derived geological profiles and inorganic/organic markers confirmed secondary NH4NO3 (27–37%, residential wood combustion (RWC; 11–51%, and diesel engine exhaust (7–22% as the dominant PM2.5 contributors. Paved road dust and de-icing materials were minor, but detectable contributors. RWC is a more important source than diesel for organic carbon (OC, but vice versa for elemental carbon (EC. A majority of secondary NH4NO3 is also attributed to RWC and diesel engines (including snow removal equipment through oxides of nitrogen (NOx emissions from these sources. Findings from this study may apply to similar situations experienced by other urban valleys.

  15. EUNIS habitat's thresholds for the Western coast of the Iberian Peninsula - A Portuguese case study

    Science.gov (United States)

    Monteiro, Pedro; Bentes, Luis; Oliveira, Frederico; Afonso, Carlos M. L.; Rangel, Mafalda O.; Gonçalves, Jorge M. S.

    2015-06-01

    The European Nature Information System (EUNIS) has been implemented for the establishment of a marine European habitats inventory. Its hierarchical classification is defined and relies on environmental variables which primarily constrain biological communities (e.g. substrate types, sea energy level, depth and light penetration). The EUNIS habitat classification scheme relies on thresholds (e.g. fraction of light and energy) which are based on expert judgment or on the empirical analysis of the above environmental data. The present paper proposes to establish and validate an appropriate threshold for energy classes (high, moderate and low) and for subtidal biological zonation (infralittoral and circalittoral) suitable for EUNIS habitat classification of the Western Iberian coast. Kinetic wave-induced energy and the fraction of photosynthetically available light exerted on the marine bottom were respectively assigned to the presence of kelp (Saccorhiza polyschides, Laminaria hyperborea and Laminaria ochroleuca) and seaweed species in general. Both data were statistically described, ordered from the largest to the smallest and percentile analyses were independently performed. The threshold between infralittoral and circalittoral was based on the first quartile while the 'moderate energy' class was established between the 12.5 and 87.5 percentiles. To avoid data dependence on sampling locations and assess the confidence interval a bootstrap technique was applied. According to this analysis, more than 75% of seaweeds are present at locations where more than 3.65% of the surface light reaches the sea bottom. The range of energy levels estimated using S. polyschides data, indicate that on the Iberian West coast the 'moderate energy' areas are between 0.00303 and 0.04385 N/m2 of wave-induced energy. The lack of agreement between different studies in different regions of Europe suggests the need for more standardization in the future. However, the obtained thresholds in

  16. Epidemiological Study of Common Dermatological Disorders in Western Nepal: A Cross-Sectional Comparative Study

    Directory of Open Access Journals (Sweden)

    Pokhrel Kumar

    2016-07-01

    Full Text Available Background: Skin diseases and their complications are a major burden on the health system of many nations. Since there is a paucity of data regarding the epidemiological status among the local population at the different parts of Nepal which are geographically distinct from each other, the present study was conducted to meet the need of an hour. Aim and Objectives: To identify and compare the pattern of dermatological diseases in the outpatients reporting to a medical camp that was organized at three distinct part of Nepal. Material and Methods: All cases with dermatological complaints attending the medical camp were included in the study after thorough examination. Data were collected, tabulated and analyzed statistically. Results: Out of 1,045 dermatological cases, who reported to the camp, 45.3% (mean age 26.3±30.3 were males and 54.7% (mean age 31.8±45.5 were females with the maximum number of disorders present in patients under the age group 16 to 30 years (33.6% for all the study groups. Non-infective conditions (55.8% outweighed infective conditions (36.8%. Conclusion: The identification of these diseases which are usually neglected by the community is important for proper diagnosis, treatment and for dermatologic education and research as well.

  17. OCULAR MANIFESTATIONS OF ATOPIC DERMATITIS- AN OBSERVATIONAL STUDY IN A TERTIARY CARE CENTRE IN WESTERN ODISHA

    Directory of Open Access Journals (Sweden)

    Swati Samikshya

    2017-08-01

    Full Text Available BACKGROUND Atopic dermatitis (AD also known as atopic eczema, is an allergic condition with hereditary predisposition. It mostly presents with intensely itchy skin, raised, splotchy lesions anywhere throughout the body. AD is most commonly seen in younger age group, the severity increases with increasing age. It is a chronic allergic condition, having both dermatologic as well as ocular manifestations. Ocular manifestations and its complications of AD are proven to be potentially morbid. The aim of this study was to evaluate the frequencies, prevalence and pattern of ocular manifestations in patients with Atopic Dermatitis in our Tertiary eye care center in Western Odisha, so that early diagnosis and treatment of symptoms can be effectively done to prevent complications. MATERIALS AND METHODS A Hospital based observational study of Ocular manifestations in 80 cases of Atopic Dermatitis was done over 12 months. To study the frequency of ocular symptoms and its complications, a study group comprising of 49 males and 31 females were examined thoroughly. Associated ocular signs, anterior segment of eye and fundus were examined. RESULTS Atopic keratoconjunctivitis (AKC having the maximum contribution (38.8% followed by severe blepharoconjunctivitis (20.3% was found in our study. Lid involvement appeared as thickening, scaling and Dennie Morgan folds, while conjunctival changes were seen in form of severe follicular and moderate to severe papillary reactions with limbal thickening. Cobblestone appearance of papillae were typically found along with papillary hypertrophy. The ocular abnormalities were mostly found in the age group of 0-10 years with an average duration of suffering from AD of >1 year. The ocular manifestations in our case group were not significantly associated with visual impairment or any serious morbidity. CONCLUSION Atopic dermatitis is a chronic allergic condition which when presents as ocular manifestations presents mostly with

  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...... to the internet, Mark Sedgwick demonstrates that the phenomenon of Western Sufism not only draws on centuries of intercultural transfers, but is also part of a long-established relationship between Western thought and Islam that can be productive, not confrontational....

  19. Maternal depression and child development: Evidence from São Paulo's Western Region Cohort Study.

    Science.gov (United States)

    Brentani, Alexandra; Fink, Günther

    2016-09-01

    While a growing body of evidence has investigated the relationship between maternal mental health and child development, evidence on children's early life outcomes remains mixed. We analyze the empirical relationship between maternal depression and children's development at age one using data from the São Paulo Western Region Cohort project. Seven hundred and ninety-eight (798) mother-child dyads living in the Butantã-Jaguaré' region of São Paulo were assessed through a home visit between January and March 2015. Maternal mental health was assessed using the Edinburgh Postnatal Depression Scale (EPDS). Mothers were classified as "possibly depressed" if their EPDS score was between 10 and 13 and as "likely depressed" if their EPDS score was > 13. The child outcomes analyzed were height, weight, and overall development as assessed by the Ages and Stages Questionnaire (ASQ). Height and weight were age-normalized using WHO growth standards. Stunting was defined as height-for-age z-score (HAZ) 2. Adjusted and unadjusted linear regression models were used to assess the associations between Edinburgh scores and child outcomes. No association was found between maternal depression variables and children's height, weight, stunting, and obesity. Positive associations were found between possible depression and ASQ (delta = 0.33; 95CI 0.11-0.54; p-valuechild development in the study setting analyzed. Further research will be needed to understand this lack of association: while it is possible that caregivers' mental health did not affect caregiving behavior, it is possible that the effect of maternal depression can vary according to timing, persistence, and intensity. It is also possible that the EPDS instrument may fail to identify mothers with clinical depression, or that children with depressed mothers get increased support from other family members or public early childhood focused programs.

  20. A core set of microsatellite markers for Western Corn Rootworm (Coleoptera: Chrysomelidae) population genetics studies

    Science.gov (United States)

    Interest in the ecological and population genetics of the western corn rootworm, Diabrotica virgifera virgifera LeConte, has grown rapidly in the last few years in North America and Europe. This interest is a result of a number of converging issues related to increasing difficult...

  1. The western Kenai Peninsula: An opportunity to study fire and its effects on soils and trees

    Science.gov (United States)

    Theresa Jain; Tara M. Barrett

    2011-01-01

    Most of the coastal Alaska inventory unit is part of the temperate rain-forest biome, and fire is an extremely rare event. However, for the western side of the Kenai Peninsula and the Cook Inlet region, fire is a common source of natural disturbance (fig. 50). Although wildfires are a normal part of the disturbance regime for this region, urban growth and associated...

  2. Tolerance of suicide, religion and suicide rates : an ecological and individual study in 19 Western countries

    NARCIS (Netherlands)

    Neeleman, J; Halpern, D; Leon, D; Lewis, G

    Background. Negative associations between religion and suicide, in individuals and countries, may be mediated by the degree to which suicide is tolerated. Methods. Linear regression was used to examine ecological associations between suicide tolerance, religion and suicide rates in 19 Western

  3. The Current State of European Studies in North America and of Scholarly Publishing in Western Europe.

    Science.gov (United States)

    Hacken, Richard

    1998-01-01

    Relates how scholarly publishing in Western Europe feeds into North America. Discusses globalization, regionalism, and particularism; new models and research methodology; Biblio-Darwinism (survival of the fittest publishing languages) and the language of the imprint; differing academic infrastructures of Europe; booming scholarly-title production;…

  4. The occultation of Surrealism: a study of the relationship between Bretonian Surrealism and western esotericism

    NARCIS (Netherlands)

    Bauduin, T.M.

    2012-01-01

    It has been said that Surrealism was nothing if not deeply involved with occultism and Western esotericism. Others claim that there was no such involvement or even that Surrealism was directly opposed to the occult and esoteric. ‘The occultation of Surrealism’ offers a fresh view of this complex and

  5. Western Michigan University: Quasi-Revolving Fund. Green Revolving Funds in Action: Case Study Series

    Science.gov (United States)

    Billingsley, Christina

    2011-01-01

    Western Michigan University has designed an innovative "Quasi-Revolving Fund" model that demonstrates the institution's full commitment to incorporating sustainability into campus operations. The Quasi-Revolving Fund recaptures money from cost-savings, similar to a typical green revolving fund, but it also sources capital from the…

  6. Descriptive study of an emergency centre in Western Kenya: Challenges and opportunities

    Directory of Open Access Journals (Sweden)

    Darlene R. House

    2014-03-01

    Conclusions: The most common presentation for acute care in western Kenya was injury related. However, the severity of illness, lack of pre-hospital transportation, and lack of community mental health services provide significant challenges and opportunities for developing ECs in sub-Saharan Africa.

  7. Life Satisfaction among Children in Different Family Structures: A Comparative Study of 36 Western Societies

    Science.gov (United States)

    Bjarnason, Thoroddur; Bendtsen, Pernille; Arnarsson, Arsaell M.; Borup, Ina; Iannotti, Ronald J.; Lofstedt, Petra; Haapasalo, Ilona; Niclasen, Birgit

    2012-01-01

    This paper examines differences in life satisfaction among children in different family structures in 36 western, industrialised countries (n = 184 496). Children living with both biological parents reported higher levels of life satisfaction than children living with a single parent or parent-step-parent. Children in joint physical custody…

  8. A study of the Pteromalidae (Hymenoptera: Chalcidoidea) from western and northwestern Iran

    OpenAIRE

    Ghahari H.; Huang J

    2012-01-01

    Pteromalids are small parasitic koinobiont wasps that grow on immature stages of other insects. Thus, they play an important role in most ecosystems, mainly as secondary or tertiary consumers. In the conducted surveys of pteromalid fauna in western and northwestern Iran, 37 parasitoids and hyperparasitoid species from 32 genera were collected and are presented here with the host records.

  9. A study of the Pteromalidae (Hymenoptera: Chalcidoidea from western and northwestern Iran

    Directory of Open Access Journals (Sweden)

    Ghahari H.

    2012-01-01

    Full Text Available Pteromalids are small parasitic koinobiont wasps that grow on immature stages of other insects. Thus, they play an important role in most ecosystems, mainly as secondary or tertiary consumers. In the conducted surveys of pteromalid fauna in western and northwestern Iran, 37 parasitoids and hyperparasitoid species from 32 genera were collected and are presented here with the host records.

  10. Intake of fatty acids in Western Europe with emphasis on trans fatty acids: The TRANSFAIR study

    NARCIS (Netherlands)

    Hulshof, K.F.A.M; Erp van - Baart, M.A.; Anttolainen, M.; Becker, W.; Church, S.M.; Couet, C.; Hermann-Kunz, E.; Kesteloot, H.; Leth, T.; Martins, I.; Moreiras, O.; Moschandreas, J.; Pizzoferrato, L.; Rimestad, A.H.; Thorgeirsdottir, H.; Amelsvoort, J.M.M. van; Aro, A.; Kafatos, A.G.; Lanzmann-Petithory, D.; Poppel, G. van

    1999-01-01

    Objective: To assess the intake of trans fatty acids (TFA) and other fatty acids in 14 Western European countries. Design and subjects: A maximum of 100 foods per country were sampled and centrally analysed. Each country calculated the intake of individual trans and other fatty acids, clusters of

  11. 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.

  12. A Literature Study on Usage of and Satisfaction Levels with Combined Treatment Including Oriental and Western Medicine

    Directory of Open Access Journals (Sweden)

    Lim Jung-Hun

    2012-09-01

    Full Text Available Objective: This study aimed to summarize and analyze the usage of and the satisfaction levels with combined treatment including Oriental and Western medicine. Methods: We searched studies on the usage of and the satisfaction levels with combined treatment including Oriental and Western medicine over the past 10 yrs (2001-2011 from 3 Korean databases (National Assembly Library, Research Information Service System, and National Discovery for Science Leaders. The reviewers also conducted a summarizing analysis by sampling the literature according to the type of study, study period, region, study subjects, sample size, type of sampling, research method, data analysis, study instruments, main results, etc. Results: When the main results of six studies on combined treatment usage and satisfaction levels were considered together, the most important decisive factor in determining the usage of combined treatment was the illness of the patient, followed by the patient’s occupation, sex, age, education, marital status, religion, treatment cost, and treatment results. In addition, the most important factor that determined satisfaction levels with combined treatment was age, followed by education, religion, income, health status, treatment procedures, staff attitude, and cleanliness. Conclusions: Elderly patients with musculoskeletal, cerebro-vascular, and circulatory system illnesses are more likely to prefer combined treatment over independent Oriental or Western treatment and are more likely to request specialized, adjusted medical care.

  13. Questioning western assessment of trauma among Tibetan torture survivors. A quantitative assessment study with comments from Buddhist Lamas

    DEFF Research Database (Denmark)

    Elsass, Peter; Carlsson, Jessica; Jespersen, Kristian

    2009-01-01

    not have an influence on the level of distress. After the assessment study, eight Tibetan lamas were interviewed about their views on our methods and results. They questioned the validity of our western rating scales and explained that our results might be influenced by the Tibetan culture, which among...... other things can be characterized as having a view and articulation of suffering much more complex than the units of our study's rating scales....

  14. Modulation of Summer Monsoon Circulation over Peninsular India by Western Ghats- A regional Climate modeling study

    Science.gov (United States)

    Rajanayagam, Lorna; Janardanan, Rajesh; Ram Mohan, H. S.

    The aim of the study is to understand the wind pattern over the Peninsular India with the modification of Orography over the region using Regional climate model. The model used in this study is the recent version (Version III) of the National Centre for Atmospheric Research (NCAR) Regional Climate Model RegCM3. The model integration is done on a horizontal resolution of 60 km. . The planetary boundary layer scheme used is that of Holtslag, cumulus parameterization scheme Emanuel of MIT, SUBEX large scale precipitation scheme and BATS ocean flux parameterization scheme. The model is run from 1st May to 30th September. The first month is taken for the spin up. The next four months are taken to study the monsoon. The simulation has been carried out for the 100%, 90%, 60% and 30% Orography (hereafter 100%o,90%o, 60%o, and 30%o) cases with RegCM3. The Zonal wind pattern for the 100%o and 90%o are similar, whereas the pattern changed for the 60% and 30%o. For the 60% and 30%o cases, the Zonal wind strengthened over the south peninsular India gradually increasing towards north. The Meridional component of the wind has a maximum over the Western Ghats between the latitudes 10o N and 15o N for the 100%o. Similar pattern has been observed for the 90%o also. For the 60%o and 30%o, the core has shifted to the northeast of India. The magnitude has decreased for the 30%o and the magnitude of meridional component is zero over the region south of 10o N. Keywords: Orography, Zonal and Meridional wind. References: Giorgi F, Mon. Wea Rev. 121: 2794 (1993) Giorgi F, Marinucci M R and Bates G T , Mon. Wea Rev. 121: 2794 (1993) K. C. Chow, Timing Liu, Johnny C. L. Chan and Yihui Ding, Int. J. Climatol. 26:1339-1359 (2006) K. C. Chow, Hang-Wai Tong and Johnny C. L. Chan, Clim. Dyn. DOI 10.1007/s00382-007- 0301-6

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

    Science.gov (United States)

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

    2014-10-01

    The detection of Campylobacter, the most commonly reported cause of foodborne gastroenteritis in the European Union, is very important for human health. The most commonly recognised risk factor for infection is the handling and/or consumption of undercooked poultry meat. The methods typically applied to evaluate the presence/absence of Campylobacter in food samples are direct plating and/or enrichment culture based on the Horizontal Method for Detection and Enumeration of Campylobacter spp. (ISO 10272-1B: 2006) and PCR. Molecular methods also allow for the detection of cells that are viable but cannot be cultivated on agar media and that decrease the time required for species identification. The current study proposes the use of two molecular methods for species identification: dot blot and PCR. The dot blot method had a sensitivity of 25 ng for detection of DNA extracted from a pure culture using a digoxigenin-labelled probe for hybridisation; the target DNA was extracted from the enrichment broth at 24 h. PCR was performed using a pair of sensitive and specific primers for the detection of Campylobacter jejuni and Campylobacter coli after 24 h of enrichment in Preston broth. The initial samples were contaminated by 5 × 10 C. jejuni cells/g and 1.5 × 10(2)C. coli cells/g, thus the number of cells present in the enrichment broth at 0 h was 1 or 3 cell/g, respectively. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. Protein kinase Cδ expression in breast cancer as measured by real-time PCR, western blotting and ELISA

    NARCIS (Netherlands)

    E. McKiernan; K. O'Brien; N. Grebenchtchikov (Nicolai); A. Geurts-Moespot (Anneke); A.M. Sieuwerts (Anieta); J.W.M. Martens (John); V. Magdolen; D. Evoy; E. McDermott; J. Crown; F.C. Sweep (Fred); M.J. Duffy (Michael)

    2008-01-01

    textabstractThe protein kinase C (PKC) family of genes encode serine/threonine kinases that regulate proliferation, apoptosis, cell survival and migration. Multiple isoforms of PKC have been described, one of which is PKCδ. Currently, it is unclear whether PKCδ is involved in promoting or inhibiting

  17. Protein kinase Cdelta expression in breast cancer as measured by real-time PCR, western blotting and ELISA.

    NARCIS (Netherlands)

    McKiernan, E.; O'Brien, K.; Grebenchtchikov, N.; Geurts-Moespot, A.; Sieuwerts, A.M.; Martens, J.W.; Magdolen, V.; Evoy, D.; McDermott, E.; Crown, J.; Sweep, F.C.; Duffy, M.J.

    2008-01-01

    The protein kinase C (PKC) family of genes encode serine/threonine kinases that regulate proliferation, apoptosis, cell survival and migration. Multiple isoforms of PKC have been described, one of which is PKCdelta. Currently, it is unclear whether PKCdelta is involved in promoting or inhibiting

  18. Cathepsin D Expression in Colorectal Cancer: From Proteomic Discovery through Validation Using Western Blotting, Immunohistochemistry, and Tissue Microarrays

    Directory of Open Access Journals (Sweden)

    Chandra Kirana

    2012-01-01

    Full Text Available Despite recent advances in surgical techniques and therapeutic treatments, survival from colorectal cancer (CRC remains disappointing with some 40–50% of newly diagnosed patients ultimately dying of metastatic disease. Current staging by light microscopy alone is not sufficiently predictive of prognosis and would benefit from additional support from biomarkers in order to stratify patients appropriately for adjuvant therapy. We have identified that cathepsin D expression was significantly greater in cells from invasive front (IF area and liver metastasis (LM than those from main tumour body (MTB. Cathepsin D expression was subsequently examined by immunohistochemistry in tissue microarrays from 119 patients with CRC. Strong expression in tumour cells at the IF did not correlate significantly with any clinico-pathological parameters examined or patient survival. However, cathepsin D expression in cells from the MTB was highly elevated in late stage CRC and showed significant correlation with subsequent distant metastasis and shorter cancer-specific survival. We also found that macrophages surrounding tumour cells stained strongly for cathepsin D but there was no significant correlation found between cathepsin D in macrophages at IF and MTB of CRC patient with the clinic-pathological parameters examined.

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

    Science.gov (United States)

    1992-09-30

    development of AIDS in subjects with hemophilia . N Engl J Med 1989;321:1141-8. 8. Margolick JB, Fauci AS. Immunopathogenesis of the acquired ...S, Taylor J. et al. Acquired immunodeficiency syndrome (AIDS)-free time after Human Immunodeficiency Virus Type I (HIV-1) seroconversion in homosexual

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

    Science.gov (United States)

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

  1. False-positive HIV-1 test results in a low-risk screening setting of voluntary blood donation. Retrovirus Epidemiology Donor Study.

    Science.gov (United States)

    Kleinman, S; Busch, M P; Hall, L; Thomson, R; Glynn, S; Gallahan, D; Ownby, H E; Williams, A E

    Persons at risk of human immunodeficiency virus 1 (HIV-1) infection, have been classified incorrectly as HIV infected because of Western blot results, but the frequency of false-positive Western blot results is unknown. To determine the frequency of false-positive HIV-1 Western blot results in US blood donors and to make projections to other screened populations. Secondarily, to validate an algorithm for evaluating possible false-positive cases. A retrospective cohort study of HIV-1 enzyme immunoassay (EIA) and Western blot results from large blood donor screening programs in which donors with suspected false-positive Western blot results underwent HIV-1 RNA polymerase chain reaction (PCR) testing and follow-up HIV-1 serology. Five US blood centers participating in the Retrovirus Epidemiology Donor Study. More than 5 million allogeneic and autologous blood donors who successfully donated blood at 1 of the 5 participating centers from 1991 through 1995. Rate of false positivity by Western blot and true HIV-1 infection status as determined by HIV-1 RNA PCR and by serologic follow-up of blood donors more than 5 weeks after donation. Of 421 donors who were positive for HIV-1 by Western blot, 39 (9.3%) met the criteria of possible false positivity because they lacked reactivity to p31. Of these, 20 (51.3%) were proven by PCR not to be infected with HIV-1. The false-positive prevalence was 4.8% of Western blot-positive donors and 0.0004% (1 in 251000) of all donors (95% confidence interval, 1 in 173000 to 1 in 379000 donors). A false diagnosis of HIV-1 infection can result from the combination of EIA and Western blot testing in blood donor and other HIV-1 screening programs. Individuals with a positive Western blot result lacking the p31 band should be counseled that, although they may be HIV infected, there is uncertainty about this conclusion. These individuals should be further evaluated by RNA PCR testing (if feasible) and HIV serologic analysis on a follow-up sample.

  2. Buddhism at Crossroads: A Case Study of Six Tibetan Buddhist Monks Navigating the Intersection of Buddhist Theology and Western Science

    Science.gov (United States)

    Sonam, Tenzin

    Recent effort to teach Western science in the Tibetan Buddhist monasteries has drawn interest both within and outside the quarters of these monasteries. This novel and historic move of bringing Western science in a traditional monastic community began around year 2000 at the behest of His Holiness the Dalai Lama, the spiritual head of Tibetan Buddhism. Despite the novelty of this effort, the literature in science education about learners from non-Western communities suggests various "cognitive conflicts" experienced by these non-Western learners due to fundamental difference in the worldview of the two knowledge traditions. Hence, in this research focuses on how six Tibetan Buddhist monks were situating/reconciling the scientific concepts like the theory of evolution into their traditional Buddhist worldview. The monks who participated in this study were engaged in a further study science at a university in the U.S. for two years. Using case study approach, the participants were interviewed individually and in groups over the two-year period. The findings revealed that although the monks scored highly on their acceptance of evolution on the Measurement of Acceptance of Theory of Evolution (MATE) survey, however in the follow-up individual and focus group interviews, certain conflicts as well as agreement between the theory of evolution and their Buddhist beliefs were revealed. The monks experienced conflicts over concepts within evolution such as common ancestry, human evolution, and origin of life, and in reconciling the Buddhist and scientific notion of life. The conflicts were analyzed using the theory of collateral learning and was found that the monks engaged in different kinds of collateral learning, which is the degree of interaction and resolution of conflicting schemas. The different collateral learning of the monks was correlated to the concepts within evolution and has no correlation to the monks' years in secular school, science learning or their

  3. Western Australia energy system

    Energy Technology Data Exchange (ETDEWEB)

    Peter Scaife; Andre Urfer; Phil Brown; Aaron Cottrell; Jason Nunn; Louis Wibberley

    2006-03-15

    The study aims to assess present and future energy supply in Western Australia, and incorporates requests made by Wesfarmers, Griffin Energy, Western Power and the Department of Industry and Resources in October 2003 to include a number of hypothetical energy futures.

  4. Western Cape Primary Care Assessment Tool (PCAT study: Measuring primary care organisation and performance in the Western Cape Province, South Africa (2013

    Directory of Open Access Journals (Sweden)

    Graham F. Bresick

    2016-03-01

    Full Text Available Background: Major health sector reform and the need for baseline measures of performance to determine impact.Aim: Baseline audit of primary healthcare (PHC performance.Setting: Cape Town and Cape Winelands (rural PHC facilities (PCFs in Western Cape Province, South Africa.Method: The South African cross-culturally validated ZA PCAT to audit PHC performance on 11 subdomains associated with improved health and reduced costs. Adult PCF users systematically sampled. All full-time doctors and nurse practitioners in PCFs sampled and all PCF managers in sub-districts sampled invited into the study.Results: Data from 1432 users, 100 clinicians and 64 managers from 13 PCFs in 10 sub-districts analysed (figures show stakeholder percentages scoring subdomain performance ‘acceptable to good’. 11.5% users scored access ‘acceptable to good’; community orientation and comprehensive services provided 20.8% and 39.9%, respectively. Total PHC score for users 50.2%; for managers and practitioners 82.8% and 88.0%, respectively. Among practitioners access was lowest (33.3%; PHC team (98.0% and comprehensive services available (100.0% highest. Among managers, access (13.5% and family centredness (45.6% are lowest; PHC team (85.9% and comprehensive services available (90.6% highest. Managers scored access, family centredness and cultural competence significantly lower than practitioners. Users scored comprehensive services available, comprehensive services provided and community orientation significantly lower than practitioners and managers.Conclusion: Gaps between users’ experience and providers’ assessments of PHC performance are identified. Features that need strengthening and alignment with best practice, provincial and national, and health policies are highlighted with implications for practitioner and manager training, health policy, and research.

  5. Western Cape Primary Care Assessment Tool (PCAT) study: Measuring primary care organisation and performance in the Western Cape Province, South Africa (2013).

    Science.gov (United States)

    Bresick, Graham F; Sayed, Abdul-Rauf; Le Grange, Cynthia; Bhagwan, Susheela; Manga, Nayna; Hellenberg, Derek

    2016-05-19

    Major health sector reform and the need for baseline measures of performance to determine impact. Baseline audit of primary healthcare (PHC) performance. Cape Town and Cape Winelands (rural) PHC facilities (PCFs) in Western Cape Province, South Africa. The South African cross-culturally validated ZA PCAT to audit PHC performance on 11 subdomains associated with improved health and reduced costs. Adult PCF users systematically sampled. All full-time doctors and nurse practitioners in PCFs sampled and all PCF managers in sub-districts sampled invited into the study. Data from 1432 users, 100 clinicians and 64 managers from 13 PCFs in 10 sub-districts analysed (figures show stakeholder percentages scoring subdomain performance 'acceptable to good'). 11.5% users scored access 'acceptable to good'; community orientation and comprehensive services provided 20.8% and 39.9%, respectively. Total PHC score for users 50.2%; for managers and practitioners 82.8% and 88.0%, respectively. Among practitioners access was lowest (33.3%); PHC team (98.0%) and comprehensive services available (100.0%) highest. Among managers, access (13.5%) and family centredness (45.6%) are lowest; PHC team (85.9%) and comprehensive services available (90.6%) highest. Managers scored access, family centredness and cultural competence significantly lower than practitioners. Users scored comprehensive services available, comprehensive services provided and community orientation significantly lower than practitioners and managers. Gaps between users' experience and providers' assessments of PHC performance are identified. Features that need strengthening and alignment with best practice, provincial and national, and health policies are highlighted with implications for practitioner and manager training, health policy, and research.

  6. Trans-Pacific Transport of Saharan Dust to Western North America: A Case Study

    Science.gov (United States)

    Kendry, Ian G. M.; Strawbridge, Kevin B.; O'Neill, Norman; Macdonald, Anne Marie; Liu, Peter S. K.; Leaitch, W. Richard; Anlauf, Kurt G.; Jaegle, Lyatt; Fairlie, T. Duncan; Westphal, Douglas L.

    2007-01-01

    The first documented case of long range transport of Saharan dust over a pathway spanning Asia and the Pacific to Western North America is described. Crustal material generated by North African dust storms during the period 28 February - 3 March 2005 reached western Canada on 13-14 March 2005 and was observed by lidar and sunphotometer in the Vancouver region and by high altitude aerosol instrumentation at Whistler Peak. Global chemical models (GEOS-CHEM and NRL NAAPS) confirm the transport pathway and suggest source attribution was simplified in this case by the distinct, and somewhat unusual, lack of dust activity over Eurasia (Gobi and Takla Makan deserts) at this time. Over western North America, the dust layer, although subsiding close to the boundary layer, did not appear to contribute to boundary layer particulate matter concentrations. Furthermore, sunphotometer observations (and associated inversion products) suggest that the dust layer had only subtle optical impact (Aerosol Optical Thickness (Tau(sub a500)) and Angstrom exponent (Alpha(sub 440-870) were 0.1 and 1.2 respectively) and was dominated by fine particulate matter (modes in aerodynamic diameter at 0.3 and 2.5microns). High Altitude observations at Whistler BC, confirm the crustal origin of the layer (rich in Ca(++) ions) and the bi-modal size distribution. Although a weak event compared to the Asian Trans-Pacific dust events of 1998 and 2001, this novel case highlights the possibility that Saharan sources may contribute episodically to the aerosol burden in western North America.

  7. The occultation of Surrealism: a study of the relationship between Bretonian Surrealism and western esotericism

    OpenAIRE

    Bauduin, T.M.

    2012-01-01

    It has been said that Surrealism was nothing if not deeply involved with occultism and Western esotericism. Others claim that there was no such involvement or even that Surrealism was directly opposed to the occult and esoteric. ‘The occultation of Surrealism’ offers a fresh view of this complex and important debate that has remained unresolved until now, and seeks to account for such differing opinions about the supposedly occult character of Surrealism, specifically under the leadership of ...

  8. Collection of population-based cancer staging information in Western Australia – a feasibility study

    Directory of Open Access Journals (Sweden)

    Katris Paul

    2005-08-01

    Full Text Available Abstract Background Routine data from cancer registries often lack information on stage of cancer, limiting their use. This study aimed to determine whether or not it is feasible to add cancer staging data to the routine data collections of a population-based Western Australian Cancer Registry (WACR. Methods For each of the five most common cancer types (prostate, colorectal, melanoma, breast and lung cancers, 60 cases were selected for staging. For the 15 next most common cancer types, 20 cases were selected. Four sources for collecting staging data were used in the following order: the WACR, the hospital based cancer registries (HBCRs, hospital medical records, and letters to treating doctors. If the case was unable to be fully staged, due to lack of information on regional lymph node invasion or distant metastases, we made the following assumptions. Cases which had data available for tumour (T and regional lymph nodes (N, but no assessment of distant metastasis (MX were assumed to have no distant metastases (M0. Cases which had data for T and M, but no assessment of regional nodal involvement (NX were assumed to have no regional nodal involvement (N0. Results The main focus of this project was the process of collecting staging data, and not the outcomes. For ovary, cervix and uterus cancers the existence of a HBCR increased the stageable proportion of cases so that staging data for these cancers could be incorporated into the WACR immediately. Breast and colorectal cancer could also be staged with adequate completeness if it were assumed that MX = M0. Similarly, melanoma and prostate cancer could be staged adequately if it were assumed that NX = N0 and MX = M0. Some cases of stomach, lung, pancreas, thyroid, testis and kidney cancers could be staged, but additional clinical input – on pathology request forms, for example – would be required to achieve useable levels of completeness. For the remaining cancer types either staging is widely

  9. Geoarchaeological studies of the Yalibirri Mindi rock shelter, Weld Range, Wajarri Yamaji Country, Western Australia

    Science.gov (United States)

    Leopold, Matthias; Brown, Viviene; Carson, Anneliese; D'Ovidio, Belinda; Yusiharni, Emielda; Winton, Vicky

    2017-04-01

    Understanding of past environmental conditions can be used to answer archaeologically based questions. Geoarchives such as soils and sediments inside rockshelters store information about chemical and physical processes from their time of formation thus allowing the reconstruction of the past. This study seeks to understand the sediment and soil formation factors at the 'Yalibirri Mindi' rock shelter, located in the Weld Range in the country of the Wajarri Yamaji Aboriginal people of Mid West Western Australia. The project is part of the 'federally funded Weld Range Web of Knowledge Project' . The aim of the work is to evaluate the origin of dated carbon material and associated sediments. Rockshelter sediments as well as two soil profiles outside the shelter were characterized using a series of different laboratory techniques such as pH and EC, nested particle sieving, ICP-OES, XRD, XRF, CN and radiocarbon analysis as well as magnetic susceptibility. An additional heating experiment was performed to simulate the influence of frequent fire on the magnetic properties of the sediments to evaluate potential anthropogenic origin of fire remains (charcoal). Pleistocene age estimates were obtained for some of the charcoal found in association with Aboriginal flaked stone artefacts. The lowest layer containing Aboriginal artefacts was dated to 29,089 ± 132 years uncal. BP providing the first evidence for Pre-Last Glacial Maximum occupation of the inland Mid West in Australia. Sediment analyses indicate that the rockshelter sediments are the result of in-situ weathering with contemporaneous human occupation rather than transport from outside. Gypsum (CaSO4.2H2O), which is not part of the natural environment in the surrounding area was identified in the rockshelter sediments and might be indicative of heating and evaporation during wood fire burning. Human induced fires had also altered the magnetic susceptibility of the sediments. Sedimentological analyses strongly support

  10. Soil tillage, physical disturbance and fauna population: a case study in western Iran

    Science.gov (United States)

    Moradi, Jabbar

    2015-04-01

    As a vital biological habitat for a great number of organisms and a medium for soil food web, soil has a great importance in regulating the two main life-supporting processes: production and decomposition. For more sustainable agricultural systems, understanding the mechanisms shaping soil fauna populations is of great importance specially in semi-arid regions with low organic matter soils. In this regard a two year study in 2008 and 2009 was conducted in western Iran to see the consequences of implementing three different tillage systems (conventional, minimum and no tillage) and three levels of organic matter amendment (0, 20 and 40 ton.ha-1 of cattle manure) over the population of soil fauna (i.e. earthworms, mite, springtail and nematodes) in three different sampling periods each year. In the second year BD decreased in the tillage treatments with mechanical turmoil but seems it started to increase in conventional tillage that can be due to higher decomposition of organic matter as the result of aeration and mixing of organic matter with the soil but shows a decrease pattern for the other two which can be due to less and no disturbance and as a result less elimination of soil aggregates. Observed earthworm populations were low besides of their patchy distribution that made the numbers unreliable to be interpreted. Soil mites showed no change regarding to treatments implemented which highlighted the importance of the need to observations in the suborder level and some other environmental variables. Soil springtails were reduced by soil tillage indicating their sensitivity to the disturbance in their physical habitat. Nematodes were mainly affected by organic matter. They showed an increase in their population (113 N.100g soil-1) in 2008 with application of 40 ton.ha-1 of cattle manure but in the second year because of the remaining effects of cattle manure the changes has been observed in response to the disturbance induced by tillage with the lowest numbers in

  11. Status of oral health awareness in Indian children as compared to Western children: A thought provoking situation (A pilot study

    Directory of Open Access Journals (Sweden)

    Grewal Navneet

    2007-03-01

    Full Text Available A study was conducted to evaluate the level of oral health awareness in a selected sample of Indian children and a selected sample of western children residing in Amritsar city of Punjab. Children were evaluated on the basis of a standardized questionnaire developed by WHO, for health awareness in children. Final results revealed the level of awareness in both the groups and the practical application of their knowledge about dental health in day to day life.

  12. Do gender-dyads have different communication patterns?: a comparative study in Western-European general practice.

    OpenAIRE

    Brink-Muinen, A. van den; Dulmen, S. van; Messerli-Rohrbach, V.; Bensing, J.

    2002-01-01

    From the viewpoint of quality of care, doctor-patient communication has become more and more important. Gender is an important factor in communication. Besides, cultural norms and values are likely to influence doctor-patient communication as well. This study examined (1) whether or not communication patterns of gender-dyads in general practice consultations differ across and between Western-European countries, and (2) if so, whether these differences continue to exist when controlling for pa...

  13. A Flash Flood Study on the Small Montaneous River Catchments in Western Romania

    Science.gov (United States)

    Győri, Maria-Mihaela; Haidu, Ionel; Humbert, Joël

    2013-04-01

    The present study focuses on flash flood modeling on several mountaneous catchments situated in Western Romania by the use of two methodologies, when rainfall and catchment characteristics are known. Hence, the Soil Conservation Service (SCS) Method and the Rational Method will be employed for the generation of the 1%, 2% and 10% historical flash flood hydrographs on the basis of data spanning from 1989-2009. The SCS Method has been applied on the three gauged catchments in the study area: Petris, Troas and Monorostia making use of the existing interconnection between GIS and the rainfall-runoff models. The DEM, soil data and land use preprocessing in GIS allowed a determination of the hydrologic parameters needed for the rainfall-runoff model, with special emphasis on determining the time of concentration, Lag time and the weighted Curve Number according to Antecedent Moisture Conditions II, adapted for the Romanian territory. HEC-HMS rainfall-runoff model (Hydrologic Engineering Center- Hydrologic Modeling System) facilitates the historical 1%, 2% and 10% flash flood hydrograph generation for the three afore mentioned watersheds. The model is calibrated against measured streamflow data from the three existing gauging stations. The results show a good match between the resulted hydrographs and the observed hydrographs under the form of the Peak Weighted Error RMS values. The hydrographs generated by surface runoff on the ungauged catchments in the area is based on an automation of a workflow in GIS, built with ArcGIS Model Builder graphical interface, as a large part of the functions needed were available as ArcGIS tools. The several components of this model calculate: the runoff depth in mm, the runoff coefficient, the travel time and finally the discharge module which is an application of the rational method, allowing the discharge computation for every cell within the catchment. The result consists of discharges for each isochrones that will be subsequently

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

    OpenAIRE

    Zhang, Pengfei; LU, HUIQI; Chen, Jia; Han,Huanxing; Ma, Wei

    2014-01-01

    Simple and sensitive detection of infectious disease at an affordable cost is urgently needed in developing nations. In this regard, the dot blot immunoassay has been used as a common protein detection method for detection of disease markers. However, the traditional signal reporting systems, such as those using enzymes or gold nanoparticles lack sensitivity and thus restrict the application of these methods for disease detection. In this study, we report a simple and sensitive detection meth...

  15. Acute lead poisoning in western Canadian cattle — A 16-year retrospective study of diagnostic case records

    Science.gov (United States)

    Cowan, Vanessa; Blakley, Barry

    2016-01-01

    This study describes the epidemiology of acute lead poisoning in western Canadian cattle over the 16-year period of 1998 to 2013 and reports background bovine tissue lead concentrations. Case records from Prairie Diagnostic Services, Western College of Veterinary Medicine, identified 525 cases of acute lead toxicity over the investigational period. Poisonings were influenced by year (P poisoned (53.5%; P poisoned. Mean toxic lead concentrations (mg/kg wet weight) in the blood, liver, and kidney were 1.30 ± 1.70 (n = 301), 33.5 ± 80.5 (n = 172), and 56.3 ± 39.7 (n = 61). Mean normal lead concentrations in the blood, liver, and kidney were 0.036 ± 0.003 mg/kg (n= 1081), 0.16 ± 0.63 mg/kg (n = 382), and 0.41 ± 0.62 mg/kg (n = 64). PMID:27041761

  16. Western conceptualizations and Eastern experience: a cross-cultural study of traumatic stress reactions among Tibetan refugees in India.

    Science.gov (United States)

    Terheggen, M A; Stroebe, M S; Kleber, R J

    2001-04-01

    This study investigated the nature and impact of traumatic experiences among Tibetan refugees in India. It explored the applicability of western conceptualizations of reactions to traumatic events among this cultural group. A randomly selected sample of refugee camp students was assessed on measures of psychological and physical complaints, and on impact as well as severity of traumatic experiences. Respondents had experienced multiple traumatic events. The majority reported ongoing health problems. More than half demonstrated symptoms of intrusion-avoidance. Those with more traumatic experiences reported more symptoms of anxiety and depression (psychologically and, particularly, somatically manifested). Cultural differences in types of experienced traumatic events and manifestations of distress are discussed, as are implications for cross-cultural trauma research using western conceptualizations and instruments.

  17. Western dietary pattern increases risk of cardiovascular disease in Iranian adults: a prospective population-based study.

    Science.gov (United States)

    Mirmiran, Parvin; Bahadoran, Zahra; Vakili, Azita Zadeh; Azizi, Fereidoun

    2017-03-01

    Limited data are available regarding the association of major dietary patterns and risk of cardiovascular disease (CVD) in Middle Eastern countries. We aimed to evaluate the association of major dietary patterns, using factor analysis, with the risk of CVD. Participants without CVD (n = 2284) were recruited from the Tehran Lipid and Glucose Study and were followed for a mean of 4.7 years. Dietary intake of participants was assessed at baseline (2006-2008); biochemical variables were evaluated at baseline and follow-up examination. Multivariate Cox proportional hazard regression models, adjusted for potential confounders, were used to estimate risk of CVD across tertiles of dietary pattern scores. Linear regression models were used to indicate association of dietary pattern scores with changes of CVD risk factors over the study period. Two major dietary patterns, Western and traditional, were identified. During a mean 4.7 ± 1.4 years of follow-up, 57 participants experienced CVD-related events. In the fully adjusted model, we observed an increased risk of CVD-related events in the highest compared to the lowest tertile category of Western dietary pattern score (HR = 2.07, 95% CI = 1.03-4.18, P for trend = 0.01). Traditional dietary pattern was not associated with incidence of CVD or CVD risk factors. A significant association was observed between the Western dietary pattern and changes in serum insulin (β = 5.88, 95% CI = 0.34-11.4). Our findings confirm that the Western dietary pattern, characterized by higher loads of processed meats, salty snacks, sweets, and soft drinks, is a dietary risk factor for CVD in the Iranian population.

  18. The Dietary Approaches to Stop Hypertension (DASH) diet, Western diet, and risk of gout in men: prospective cohort study

    Science.gov (United States)

    Rai, Sharan K; Fung, Teresa T; Lu, Na; Keller, Sarah F; Curhan, Gary C

    2017-01-01

    Objective To prospectively examine the relation between the Dietary Approaches to Stop Hypertension (DASH) and Western diets and risk of gout (ie, the clinical endpoint of hyperuricemia) in men. Design Prospective cohort study. Setting The Health Professionals Follow-up Study. Participants 44 444 men with no history of gout at baseline. Using validated food frequency questionnaires, each participant was assigned a DASH dietary pattern score (based on high intake of fruits, vegetables, nuts and legumes, low fat dairy products, and whole grains, and low intake of sodium, sweetened beverages, and red and processed meats) and a Western dietary pattern score (based on high intake of red and processed meats, French fries, refined grains, sweets, and desserts). Main outcome measure Risk of incident gout meeting the preliminary American College of Rheumatology survey criteria for gout, adjusting for potential confounders, including age, body mass index, hypertension, diuretic use, and alcohol intake. Results During 26 years of follow-up, 1731 confirmed cases of incident gout were documented. A higher DASH dietary pattern score was associated with a lower risk for gout (adjusted relative risk for extreme fifths 0.68, 95% confidence interval 0.57 to 0.80, P value for trend <0.001). In contrast, a higher Western dietary pattern score was associated with an increased risk for gout (1.42, 1.16 to 1.74, P=0.005). Conclusion The DASH diet is associated with a lower risk of gout, suggesting that its effect of lowering uric acid levels in individuals with hyperuricemia translates to a lower risk of gout. Conversely, the Western diet is associated with a higher risk of gout. The DASH diet may provide an attractive preventive dietary approach for men at risk of gout. PMID:28487277

  19. A new centennial index to study the Western North Pacific Monsoon decadal variability

    Science.gov (United States)

    Vega, Inmaculada; Gómez-Delgado, F. de Paula; Gallego, David; Ribera, Pedro; Peña-Ortiz, Cristina; García-Herrera, Ricardo

    2016-04-01

    The concept of the Western North Pacific Summer Monsoon (WNPSM) appeared for the first time in 1987. It is, unlike the Indian Summer Monsoon (ISM) and the East Asian summer monsoon (EASM), an oceanic monsoon mostly driven by the meridional gradient of sea surface temperature. Its circulation is characterized by a northwest-southeast oriented monsoon trough with intense precipitation and low-level southwesterlies and upper-tropospheric easterlies in the region [100°-130° E, 5°-15°N]. Up to now, the primary index to characterize the WNPSM has been the Western North Pacific Monsoon Index (WNPMI) which covers the 1949-2013 period. The original WNPMI was defined as the difference of 850-hPa westerlies between two regions: D1 [5°-15°N, 100°-130°E] and D2 [20°-30°N, 110°-140°E]. Both domains are included in the main historical ship routes circumnavigating Asia for hundreds of years. Many of the logbooks of these ships have been preserved in historical archives and they usually contain daily observations of wind force and direction. Therefore, it has been possible to compute a new index of instrumental character, which reconstructs the WNPSM back to the middle of the 19th Century, by using solely historical wind direction records preserved in logbooks. We define the monthly Western North Pacific Directional Index (WNPDI) as the sum of the persistence of the low-level westerly winds in D1 and easterly winds in D2. The advantages of this new index are its nature (instrumental) and its length (1849-2013), which is 100 years longer than the WNPMI (which was based on reanalysis data). Our WNPDI shows a high correlation (r=+0.87, pCompetitividad through the project INCITE (CGL2013-44530-P, BES-2014-069733).

  20. Past climate change and plant evolution in Western North America: a case study in Rosaceae.

    Directory of Open Access Journals (Sweden)

    Mats Töpel

    Full Text Available Species in the ivesioid clade of Potentilla (Rosaceae are endemic to western North America, an area that underwent widespread aridification during the global temperature decrease following the Mid-Miocene Climatic Optimum. Several morphological features interpreted as adaptations to drought are found in the clade, and many species occupy extremely dry habitats. Recent phylogenetic analyses have shown that the sister group of this clade is Potentilla section Rivales, a group with distinct moist habitat preferences. This has led to the hypothesis that the ivesioids (genera Ivesia, Horkelia and Horkeliella diversified in response to the late Tertiary aridification of western North America. We used phyloclimatic modeling and a fossil-calibrated dated phylogeny of the family Rosaceae to investigate the evolution of the ivesioid clade. We have combined occurrence- and climate data from extant species, and used ancestral state reconstruction to model past climate preferences. These models have been projected into paleo-climatic scenarios in order to identify areas where the ivesioids may have occurred. Our analysis suggests a split between the ivesioids and Potentilla sect. Rivales around Late Oligocene/Early Miocene (∼23 million years ago, Ma, and that the ivesioids then diversified at a time when summer drought started to appear in the region. The clade is inferred to have originated on the western slopes of the Rocky Mountains from where a westward range expansion to the Sierra Nevada and the coast of California took place between ∼12-2 Ma. Our results support the idea that climatic changes in southwestern North America have played an important role in the evolution of the local flora, by means of in situ adaptation followed by diversification.

  1. Case study: Examples of Wooden Vernacular Architecture - Turkish Houses in Western Anatolia

    Directory of Open Access Journals (Sweden)

    Yilmaz Karaman Özgül

    2015-12-01

    Full Text Available Wooden material is described as the oldest construction material along with stone. In the historical process, especially for the buildings of shelter, timber construction components are seen to be used as primary and indispensable structural system components in different geographies of the world as well as Anatolia. This paper focuses on timber-frame structures of four different settlements from Western Anatolia. It is aimed to analyze the traditional techniques, in order to transmit our cultural heritage to the next generations, as well as obtaining some data on construction techniqes to be used for creating alternative solutions to the house production of today.

  2. The evolution of Eastern Tornquist-Paleoasian Ocean and subsequent continental collisions: A case study from the Western Tatra Mountains, Central Western Carpathians (Poland)

    OpenAIRE

    Aleksandra Gawęda; Jolanta Burda; Jan Golonka; Urs Klötzli; David Chew; Krzysztof Szopa; Michael Wiedenbeck

    2017-01-01

    The crystalline basement of the Tatra Mountains in the Central Western Carpathians, forms part of the European Variscides and contains fragments of Gondwanan provenance. Metabasite rocks of MORB affinity in the Tatra Mountains are represented by two suites of amphibolites present in two metamorphic units (the Ornak and Goryczkowa Units) intercalated with metapelitic rocks. They are interpreted as relics of ocean crust, with zircon δ18OVSMOW values of 4.97–6.96‰. Zircon REE patterns suggest ox...

  3. Comparative study on the impact of coal and uranium mining, processing, and transportation in the western United States

    Energy Technology Data Exchange (ETDEWEB)

    Sandquist, G.M.

    1979-06-01

    A comparative study and quantitative assessment of the impacts, costs and benefits associated with the mining, processing and transportation of coal and uranium within the western states, specifically Arizona, California, Colorado, Montana, New Mexico, Oregon, Utah, Washington and Wyoming are presented. The western states possess 49% of the US reserve coal base, 67% of the total identified reserves and 82% of the hypothetical reserves. Western coal production has increased at an average annual rate of about 22% since 1970 and should become the major US coal supplier in the 1980's. The Colorado Plateau (in Arizona, Colorado, New Mexico and Utah) and the Wyoming Basin areas account for 72% of the $15/lb U/sub 3/O/sub 8/ resources, 76% of the $30/lb, and 75% of the $50/lb resources. It is apparent that the West will serve as the major supplier of domestic US coal and uranium fuels for at least the next several decades. Impacts considered are: environmental impacts, (land, water, air quality); health effects of coal and uranium mining, processing, and transportation; risks from transportation accidents; radiological impact of coal and uranium mining; social and economic impacts; and aesthetic impacts (land, air, noise, water, biota, and man-made objects). Economic benefits are discussed.

  4. No official identity: a data linkage study of birth registration of Aboriginal children in Western Australia.

    Science.gov (United States)

    Gibberd, Alison J; Simpson, Judy M; Eades, Sandra J

    2016-08-01

    Evidence of identity, particularly a birth certificate, is essential to access many rights. However, the births of many Aboriginal Australians are not registered when they are infants. We examined factors related to birth registration among Western Australian children born to Aboriginal mothers. All births to Aboriginal mothers in the Midwives Notification System in Western Australia (WA) from 1980 to 2010 were linked to birth registrations. Associations between registration and maternal and child characteristics were examined for children aged under 16 years in 2012. Among 49,694 births between 1980 and 2010, 18% of those aged under 16 years had unregistered births, compared to 3% of those aged 16-32 years. Unregistered births were most strongly associated with young maternal age at first birth (adjusted odds ratio [AOR] 5.22; 95%CI 3.07-8.86; for 16 years or younger vs 30 years or older, among non-smokers), remoteness (AOR 2.17; 95%CI 1.87-2.52; very remote vs major cities), mothers whose own birth was unregistered (AOR 3.00; 95%CI 1.78-5.07) and no private hospital insurance (AOR 0.19; 95%CI 0.11-0.31; insured vs uninsured). Unregistered births are common among WA Aboriginal children, particularly in disadvantaged families. Assistance before discharge from hospital may increase birth registrations. © 2016 Public Health Association of Australia.

  5. Sensitivity of prestaining RNA with ethidium bromide before electrophoresis and performance of subsequent northern blots using heterologous DNA probes.

    Science.gov (United States)

    Zhao, Yun; Du, Linfang; Zhang, Nianhui

    2013-06-01

    Adding ethidium bromide (EtBr) at low concentrations to RNA samples before running formaldehyde-agarose gels affords the advantages of checking RNA integrity and evaluating the quality of size-separation at any time during electrophoresis or immediately after either electrophoresis or blotted the separated RNA onto the membrane without significantly compromising mobility, transfer, or hybridization. In this study, we systematically examined the factors that affect the sensitivity of RNA prestaining by heating RNA samples that include EtBr before electrophoresis under different denaturation conditions. We also examined the efficiency of the hybridization of EtBr-prestained RNA with heterologous DNA probes. The results showed that the fluorescent intensity of EtBr-prestained RNA was affected not only by the EtBr concentration as previously reported but also by the RNA amount, denaturation time, and denaturation temperature. Prior staining of RNA with 40 μg/mL EtBr significantly decreased the efficiency of Northern blot hybridization with heterologous DNA probes. We propose that to best combine staining sensitivity and the efficiency of Northern blot hybridization with heterologous DNA probes, the concentration of EtBr used to prestain RNA should not exceed 30 μg/mL. The efficiency of the hybridization of EtBr-prestained RNA was affected not only by factors that affect staining sensitivity but also by the type of probe used.

  6. Groundwater quality in the Western San Joaquin Valley study unit, 2010: California GAMA Priority Basin Project

    Science.gov (United States)

    Fram, Miranda S.

    2017-06-09

    Water quality in groundwater resources used for public drinking-water supply in the Western San Joaquin Valley (WSJV) was investigated by the USGS in cooperation with the California State Water Resources Control Board (SWRCB) as part of its Groundwater Ambient Monitoring and Assessment (GAMA) Program Priority Basin Project. The WSJV includes two study areas: the Delta–Mendota and Westside subbasins of the San Joaquin Valley groundwater basin. Study objectives for the WSJV study unit included two assessment types: (1) a status assessment yielding quantitative estimates of the current (2010) status of groundwater quality in the groundwater resources used for public drinking water, and (2) an evaluation of natural and anthropogenic factors that could be affecting the groundwater quality. The assessments characterized the quality of untreated groundwater, not the quality of treated drinking water delivered to consumers by water distributors.The status assessment was based on data collected from 43 wells sampled by the U.S. Geological Survey for the GAMA Priority Basin Project (USGS-GAMA) in 2010 and data compiled in the SWRCB Division of Drinking Water (SWRCB-DDW) database for 74 additional public-supply wells sampled for regulatory compliance purposes between 2007 and 2010. To provide context, concentrations of constituents measured in groundwater were compared to U.S. Environmental Protection Agency (EPA) and SWRCB-DDW regulatory and non-regulatory benchmarks for drinking-water quality. The status assessment used a spatially weighted, grid-based method to estimate the proportion of the groundwater resources used for public drinking water that has concentrations for particular constituents or class of constituents approaching or above benchmark concentrations. This method provides statistically unbiased results at the study-area scale within the WSJV study unit, and permits comparison of the two study areas to other areas assessed by the GAMA Priority Basin Project

  7. The metabolic signature associated with the Western dietary pattern: a cross-sectional study.

    Science.gov (United States)

    Bouchard-Mercier, Annie; Rudkowska, Iwona; Lemieux, Simone; Couture, Patrick; Vohl, Marie-Claude

    2013-12-11

    Metabolic profiles have been shown to be associated to obesity status and insulin sensitivity. Dietary intakes influence metabolic pathways and therefore, different dietary patterns may relate to modifications in metabolic signatures. The objective was to verify associations between dietary patterns and metabolic profiles composed of amino acids (AAs) and acylcarnitines (ACs). 210 participants were recruited in the greater Quebec City area between September 2009 and December 2011. Dietary patterns had been previously derived using principal component analysis (PCA). The Prudent dietary pattern was characterised by higher intakes of vegetables, fruits, whole grain products, non-hydrogenated fat and lower intakes of refined grain products, whereas the Western dietary pattern was associated with higher intakes of refined grain products, desserts, sweets and processed meats. Targeted metabolites were quantified in 37 participants with the Biocrates Absolute IDQ p150 (Biocrates Life Sciences AG, Austria) mass spectrometry method (including 14 amino acids and 41 acylcarnitines). PCA analysis with metabolites including AAs and ACs revealed two main components explaining the most variance in overall data (13.8%). PC1 was composed mostly of medium- to long-chain ACs (C16:2, C14:2, C14:2-OH, C16, C14:1-OH, C14:1, C10:2, C5-DC/C6-OH, C12, C18:2, C10, C4:1-DC/C6, C8:1 and C2) whereas PC2 included certain AAs and short-chain ACs (xLeu, Met, Arg, Phe, Pro, Orn, His, C0, C3, C4 and C5). The Western dietary pattern correlated negatively with PC1 and positively with PC2 (r = -0.34, p = 0.05 and r = 0.38, p = 0.03, respectively), independently of age, sex and BMI. These findings suggest that the Western dietary pattern is associated with a specific metabolite signature characterized by increased levels of AAs including branched-chain AAs (BCAAs) and short-chain ACs.

  8. The metabolic signature associated with the Western dietary pattern: a cross-sectional study

    Science.gov (United States)

    2013-01-01

    Background Metabolic profiles have been shown to be associated to obesity status and insulin sensitivity. Dietary intakes influence metabolic pathways and therefore, different dietary patterns may relate to modifications in metabolic signatures. The objective was to verify associations between dietary patterns and metabolic profiles composed of amino acids (AAs) and acylcarnitines (ACs). Methods 210 participants were recruited in the greater Quebec City area between September 2009 and December 2011. Dietary patterns had been previously derived using principal component analysis (PCA). The Prudent dietary pattern was characterised by higher intakes of vegetables, fruits, whole grain products, non-hydrogenated fat and lower intakes of refined grain products, whereas the Western dietary pattern was associated with higher intakes of refined grain products, desserts, sweets and processed meats. Targeted metabolites were quantified in 37 participants with the Biocrates Absolute IDQ p150 (Biocrates Life Sciences AG, Austria) mass spectrometry method (including 14 amino acids and 41 acylcarnitines). Results PCA analysis with metabolites including AAs and ACs revealed two main components explaining the most variance in overall data (13.8%). PC1 was composed mostly of medium- to long-chain ACs (C16:2, C14:2, C14:2-OH, C16, C14:1-OH, C14:1, C10:2, C5-DC/C6-OH, C12, C18:2, C10, C4:1-DC/C6, C8:1 and C2) whereas PC2 included certain AAs and short-chain ACs (xLeu, Met, Arg, Phe, Pro, Orn, His, C0, C3, C4 and C5). The Western dietary pattern correlated negatively with PC1 and positively with PC2 (r = −0.34, p = 0.05 and r = 0.38, p = 0.03, respectively), independently of age, sex and BMI. Conclusion These findings suggest that the Western dietary pattern is associated with a specific metabolite signature characterized by increased levels of AAs including branched-chain AAs (BCAAs) and short-chain ACs. Trial registration NCT01343342 PMID:24330454

  9. Eastward Expansion of Western Learning: A Study of Westernisation of China's Modern Education by Chinese Government Overseas-Study Scholarships

    Science.gov (United States)

    Lin, Ren-Jie Vincent

    2016-01-01

    This article aims to trace back the history of how Chinese Government attempted to strengthen its national power by learning from the USA, Western Europe and Japan since the mid-nineteenth century, as well as to analyse the influences Westernisation had on the development of China's modern education. In this process, the Chinese Government…

  10. Western Canada study of animal health effects associated with exposure to emissions from oil and natural gas field facilities. Study design and data collection II. Location of study herds relative to the oil and gas industry in Western Canada.

    Science.gov (United States)

    Waldner, Cheryl L

    2008-01-01

    During the late part of 2000 and early months of 2001, project veterinarians recruited 205 beef herds to participate in a study of the effects of emissions from the upstream oil and gas industry on cattle reproduction and health. Researchers developed herd-selection criteria to optimize the range of exposure to facilities, including oil and gas wells, battery sites, and gas-gathering and gas-processing facilities across the major cattle-producing areas of Western Canada. Herds were initially selected on the basis of a ranking system of exposure potential on the basis of herd-owner reports of the locations of their operations in relation to oil and gas industry facilities. At the end of the study, researchers summarized data obtained from provincial regulatory agencies on facility location and reported flaring and venting volumes for each herd and compared these data to the original rankings of herd-exposure potential. Through this selection process, the researchers were successful in obtaining statistically significant differences in exposure to various types of oil and gas facility types and reported emissions among herds recruited for the study.

  11. Rural perspectives of climate change: a study from Saurastra and Kutch of Western India.

    Science.gov (United States)

    Moghariya, Dineshkumar P; Smardon, Richard C

    2014-08-01

    This research reports on rural people's beliefs and understandings of climate change in the Saurastra/ Kutch region of Western India. Results suggest that although most rural respondents have not heard about the scientific concept of climate change, they have detected changes in the climate. They appear to hold divergent understandings about climate change and have different priorities for causes and solutions. Many respondents appear to base their understandings of climate change upon a mix of ideas drawn from various sources and rely on different kinds of reasoning in relation to both causes of and solutions to climate change to those used by scientists. Environmental conditions were found to influence individuals' understanding of climate change, while demographic factors were not. The results suggest a need to learn more about people's conceptual models and understandings of climate change and a need to include local climate research in communication efforts.

  12. Intake of fatty acids in Western Europe with emphasis on trans fatty acids: The TRANSFAIR study

    DEFF Research Database (Denmark)

    Hulshof, K. F. A. M.; Erp-Baart, M. A. van; Anttolainen, M.

    1999-01-01

    Objective: To assess the intake of trans fatty acids (TFA) and other fatty acids in 14 Western European countries. Design and subjects: A maximum of 100 foods per country were sampled and centrally analysed. Each country calculated the intake of individual trans and other fatty acids, clusters...... of fatty acids and total fat in adults and/or the total population using the best available national food consumption data set. Results: A wide variation was observed in the intake of total fat and (clusters) of fatty acids in absolute amounts. The variation in proportion of energy derived from total fat...... and from clusters of fatty acids was less. Only in Finland, Italy, Norway and Portugal total fat did provide on average less than 35% of energy intake. Saturated fatty acids (SFA) provided on average between 10% and 19% of total energy intake, with the lowest contribution in most Mediterranean countries...

  13. Western Dietary Pattern Increases, and Prudent Dietary Pattern Decreases, Risk of Incident Diverticulitis in a Prospective Cohort Study.

    Science.gov (United States)

    Strate, Lisa L; Keeley, Brieze R; Cao, Yin; Wu, Kana; Giovannucci, Edward L; Chan, Andrew T

    2017-04-01

    Dietary fiber is implicated as a risk factor for diverticulitis. Analyses of dietary patterns may provide information on risk beyond those of individual foods or nutrients. We examined whether major dietary patterns are associated with risk of incident diverticulitis. We performed a prospective cohort study of 46,295 men who were free of diverticulitis and known diverticulosis in 1986 (baseline) using data from the Health Professionals Follow-Up Study. Each study participant completed a detailed medical and dietary questionnaire at baseline. We sent supplemental questionnaires to men reporting incident diverticulitis on biennial follow-up questionnaires. We assessed diet every 4 years using a validated food frequency questionnaire. Western (high in red meat, refined grains, and high-fat dairy) and prudent (high in fruits, vegetables, and whole grains) dietary patterns were identified using principal component analysis. Follow-up time accrued from the date of return of the baseline questionnaire in 1986 until a diagnosis of diverticulitis, diverticulosis or diverticular bleeding; death; or December 31, 2012. The primary end point was incident diverticulitis. During 894,468 person years of follow-up, we identified 1063 incident cases of diverticulitis. After adjustment for other risk factors, men in the highest quintile of Western dietary pattern score had a multivariate hazard ratio of 1.55 (95% CI, 1.20-1.99) for diverticulitis compared to men in the lowest quintile. High vs low prudent scores were associated with decreased risk of diverticulitis (multivariate hazard ratio, 0.74; 95% CI, 0.60-0.91). The association between dietary patterns and diverticulitis was predominantly attributable to intake of fiber and red meat. In a prospective cohort study of 46,295 men, a Western dietary pattern was associated with increased risk of diverticulitis, and a prudent pattern was associated with decreased risk. These data can guide dietary interventions for the prevention of

  14. Application of Monoclonal Antibodies against Bioactive Natural Products: Eastern Blotting and Preparation of Knockout Extract

    Directory of Open Access Journals (Sweden)

    Hiroyuki Tanaka

    2012-01-01

    Full Text Available Matrix-assisted laser desorption/ionization (MALDI tof mass spectrometry was used for the confirmation of hapten number in synthesized antigen. As application of MAb, the MAbs against ginsenosides and glycyrrhizin have been prepared resulting in the development of two new techniques that we named the eastern blotting method and the knockout extract preparation. In eastern blotting technique, glycosides like ginsenosides and glycyrrhizin separated by silica gel TLC were blotted to PVDF membrane that was treated with a NaIO4 solution followed by BSA resulted in glycoside-BSA conjugate on a PVDF membrane. The blotted spots were stained by MAb. Double staining of eastern blotting for ginsenosides using antiginsenoside Rb1 and Rg1 MAbs promoted complete identification of ginsenosides in Panax species. The immunoaffinity concentration of glycyrrhizin was determined by immunoaffinity column conjugated with antiglycyrrhizin MAb resulting in the glycyrrhizin-knockout extract, which was determined by the synergic effect with glycyrrhizin on NO production using the cell line.

  15. Examining the safety of menstrual cups among rural primary school girls in western Kenya: observational studies nested in a randomised controlled feasibility study.

    OpenAIRE

    Juma, Jane; Nyothach, Elizabeth; Laserson, Kayla F; Oduor, Clifford; Arita, Lilian; Ouma, Caroline; Oruko, Kelvin; Omoto, Jackton; Mason, Linda; Alexander, Kelly; Fields, Barry; Onyango, Clayton; Phillips-Howard, Penelope

    2017-01-01

    Objective Examine the safety of menstrual cups against sanitary pads and usual practice in Kenyan schoolgirls. Design Observational studies nested in a cluster randomised controlled feasibility study. Setting 30 primary schools in a health and demographic surveillance system in rural western Kenya. Participants Menstruating primary schoolgirls aged 14?16 years participating in a menstrual feasibility study. Interventions Insertable menstrual cup, monthly sanitary pads or ?usual practice? (con...

  16. Rockfall risk evaluation using geotechnical survey, remote sensing data, and GIS: a case study from western Greece

    Science.gov (United States)

    Nikolakopoulos, Konstantinos; Depountis, Nikolaos; Vagenas, Nikolaos; Kavoura, Katerina; Vlaxaki, Eleni; Kelasidis, George; Sabatakakis, Nikolaos

    2015-06-01

    In this paper a specific example of the synergistic use of geotechnical survey, remote sensing data and GIS for rockfall risk evaluation is presented. The study area is located in Western Greece. Extensive rockfalls have been recorded along Patras - Ioannina highway just after the cable-stayed bridge of Rio-Antirrio, at Klokova site. The rockfalls include medium- sized limestone boulders with volume up to 1.5m3. A detailed engineering geological survey was conducted including rockmass characterization, laboratory testing and geological - geotechnical mapping. Many Rockfall trajectory simulations were done. Rockfall risk along the road was estimated using spatial analysis in a GIS environment.

  17. Contributions to the phytosociological study of Sessile and Turkey oak forests in the Orastie area (Central-Western Romania)

    OpenAIRE

    Petru BURESCU; Valeriu Ioan VINTAN

    2012-01-01

    In the current paper we present a phytosociological study of the phytocoenoses of the association Quercetum petraeae-cerris Soó 1963 (Syn.: Quercetum polycarpae-cerris G. Popescu 1988) identified in the Orăştie river basin, situated in the central-western part of Romania.The characterisation of the association under analysis as well as the presentation of the phytosociological table have been done by selecting the most representative relevés performed in the Turkey and Sessile oak forests of...

  18. Cruise and Data Report of USA-PRC Joint Air-Sea Interaction Studies in the Western Pacific Ocean (NODC Accession 8700374)

    Data.gov (United States)

    National Oceanic and Atmospheric Administration, Department of Commerce — The USA-PRC Joint Program on Air-Sea Interaction Studies in the Tropical Western Pacific is a component of the Protocol on Cooperation in the Field of Marine and...

  19. Acceptability of Male Circumcision among College Students in Medical Universities in Western China: A Cross-Sectional Study.

    Directory of Open Access Journals (Sweden)

    Junjun Jiang

    Full Text Available Male circumcision (MC has been shown to reduce the risk of female to male transmission of HIV. The goal of this survey was to explore MC's acceptability and the factors associated with MC among college students in medical universities in western China.A cross-sectional study was carried out in three provinces in western China (Guangxi, Chongqing and Xinjiang to assess the acceptability of MC as well as to discover factors associated with the acceptability among college students in medical universities. A total of 1,790 uncircumcised male students from three medical universities were enrolled in this study. In addition, 150 students who had undergone MC were also enrolled in the survey, and they participated in in-depth interviews.Of all the uncircumcised participants (n = 1,790, 55.2% (n = 988 were willing to accept MC. Among those who accepted MC, 67.3% thought that MC could improve their sexual partners' hygiene, 46.3% believed that HIV and sexually transmitted diseases (STDs could be partially prevented by MC. The multivariable logistic regression indicates that MC's acceptability was associated with three factors: the redundant foreskin (OR = 10.171, 95% CI = 7.629-13.559, knowing the hazard of having a redundant foreskin (OR = 1.597, 95% CI = 1.097-2.323, and enhancing sexual pleasure (OR = 1.628, 95% CI = 1.312-2.021. The in-depth interviews for subjects who had undergone MC showed that the major reason for having MC was the redundant foreskin (87.3%, followed by the benefits and the fewer complications of having MC done. In addition, most of these participants (65.3% said that the MC could enhance sexual satisfaction.MC's acceptance among college students in medical universities is higher than it is among other populations in western China. An implementation of an MC programme among this population is feasible in the future.

  20. Acceptability of Male Circumcision among College Students in Medical Universities in Western China: A Cross-Sectional Study.

    Science.gov (United States)

    Jiang, Junjun; Su, Jinming; Yang, Xiaobo; Huang, Mingbo; Deng, Wei; Huang, Jiegang; Liang, Bingyu; Qin, Bo; Upur, Halmurat; Zhong, Chaohui; Wang, Qianqiu; Wang, Qian; Ruan, Yuhua; Ye, Li; Liang, Hao

    2015-01-01

    Male circumcision (MC) has been shown to reduce the risk of female to male transmission of HIV. The goal of this survey was to explore MC's acceptability and the factors associated with MC among college students in medical universities in western China. A cross-sectional study was carried out in three provinces in western China (Guangxi, Chongqing and Xinjiang) to assess the acceptability of MC as well as to discover factors associated with the acceptability among college students in medical universities. A total of 1,790 uncircumcised male students from three medical universities were enrolled in this study. In addition, 150 students who had undergone MC were also enrolled in the survey, and they participated in in-depth interviews. Of all the uncircumcised participants (n = 1,790), 55.2% (n = 988) were willing to accept MC. Among those who accepted MC, 67.3% thought that MC could improve their sexual partners' hygiene, 46.3% believed that HIV and sexually transmitted diseases (STDs) could be partially prevented by MC. The multivariable logistic regression indicates that MC's acceptability was associated with three factors: the redundant foreskin (OR = 10.171, 95% CI = 7.629-13.559), knowing the hazard of having a redundant foreskin (OR = 1.597, 95% CI = 1.097-2.323), and enhancing sexual pleasure (OR = 1.628, 95% CI = 1.312-2.021). The in-depth interviews for subjects who had undergone MC showed that the major reason for having MC was the redundant foreskin (87.3%), followed by the benefits and the fewer complications of having MC done. In addition, most of these participants (65.3%) said that the MC could enhance sexual satisfaction. MC's acceptance among college students in medical universities is higher than it is among other populations in western China. An implementation of an MC programme among this population is feasible in the future.

  1. The Dietary Approaches to Stop Hypertension (DASH) diet, Western diet, and risk of gout in men: prospective cohort study.

    Science.gov (United States)

    Rai, Sharan K; Fung, Teresa T; Lu, Na; Keller, Sarah F; Curhan, Gary C; Choi, Hyon K

    2017-05-09

    Objective To prospectively examine the relation between the Dietary Approaches to Stop Hypertension (DASH) and Western diets and risk of gout (ie, the clinical endpoint of hyperuricemia) in men.Design Prospective cohort study.Setting The Health Professionals Follow-up Study.Participants 44 444 men with no history of gout at baseline. Using validated food frequency questionnaires, each participant was assigned a DASH dietary pattern score (based on high intake of fruits, vegetables, nuts and legumes, low fat dairy products, and whole grains, and low intake of sodium, sweetened beverages, and red and processed meats) and a Western dietary pattern score (based on high intake of red and processed meats, French fries, refined grains, sweets, and desserts).Main outcome measure Risk of incident gout meeting the preliminary American College of Rheumatology survey criteria for gout, adjusting for potential confounders, including age, body mass index, hypertension, diuretic use, and alcohol intake.Results During 26 years of follow-up, 1731 confirmed cases of incident gout were documented. A higher DASH dietary pattern score was associated with a lower risk for gout (adjusted relative risk for extreme fifths 0.68, 95% confidence interval 0.57 to 0.80, P value for trend gout (1.42, 1.16 to 1.74, P=0.005).Conclusion The DASH diet is associated with a lower risk of gout, suggesting that its effect of lowering uric acid levels in individuals with hyperuricemia translates to a lower risk of gout. Conversely, the Western diet is associated with a higher risk of gout. The DASH diet may provide an attractive preventive dietary approach for men at risk of gout. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://group.bmj.com/group/rights-licensing/permissions.

  2. Geographic factors as determinants of food security: a Western Australian food pricing and quality study.

    Science.gov (United States)

    Pollard, Christina Mary; Landrigan, Timothy John; Ellies, Pernilla Laila; Kerr, Deborah Anne; Lester, Matthew Langdon Underwood; Goodchild, Stanley Edward

    2014-01-01

    Food affordability and quality can influence food choice. This research explores the impact of geographic factors on food pricing and quality in Western Australia (WA). A Healthy Food Access Basket (HFAB) was cost and a visual and descriptive quality assessment of 13 commonly consumed fresh produce items was conducted in-store on a representative sample of 144 food grocery stores. The WA retail environment in 2010 had 447 grocery stores servicing 2.9 million people: 38% of stores the two major chains (Coles® Supermarkets Australia and Woolworths ® Limited) in population dense areas, 50% were smaller independently owned stores (Independent Grocers Association®) in regional areas as well, and 12% Indigenous community stores in very remote areas. The HFAB cost 24% (pfoods cost more and the quality of fresh produce was lower. Food affordability and quality may deter healthier food choice in geographically isolated communities. Improving affordability and quality of nutritious foods in remote communities may positively impact food choices, improve food security and prevent diet-sensitive chronic disease. Policy makers should consider influencing agriculture, trade, commerce, transport, freight, and modifying local food economies.

  3. Aphasia following left thalamic hemorrhage. A study by Western Aphasia Battery and single photon emission CT

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    Makishita, Hideo; Miyasaka, Motomaro; Tanizaki, Yoshio; Yanagisawa, Nobuo; Sugishita, Morihiro

    1984-07-01

    A report is given of 7 patients with left thalamic hemorrhage in the chronic stage (from 1.5 months to 4.5 months) in which language disorders were examined by Western Aphasia Battery (WAB) and cerebral blood flow was measured by single photon emission CT. Examination of language by WAB revealed 4 aphasics out of 7 cases, and 3 patients had no language deficit. The patient with Wernicke's aphasia showed low density area only in the left posterior thalamus in X-ray CT, and revealed severe low blood flow area extending to left temporal lobe in emission CT. In the case with transcortical sensory aphasia, although X-ray CT showed no obvious low density area, emission CT revealed moderate low flow area in the left temporooccipital region and low blood flow at the left thalamus. In one of the two patients classified as anomic aphasia, emission CT showed slight low flow area at the temporo-occipital region similar to the case with transcortical sensory aphasia. In another case with anomic aphasia there was a wide low density area all over the left thalamus and midline shift to the right in X-ray CT, and emission CT showed severe low blood flow in the same region spreading widely toward the cerebral surface. In all of the 3 patients without aphasia, emission CT showed low flow region restricted to the left thalamus.

  4. Drivers of sustained hygiene behaviour change: A case study from mid-western Nepal.

    Science.gov (United States)

    McMichael, Celia; Robinson, Priscilla

    2016-08-01

    Behaviour change is central to the prevention of many population health problems, yet it is typically difficult to initiate and sustain. This paper reports on an evaluation of a water, sanitation and hygiene (WASH) intervention in mid-western Nepal, with particular focus on the drivers and barriers for handwashing with soap/ash and elimination of open defecation. The research was conducted during October-November 2014, two and half years following the intervention's end-point. Qualitative data were collected from the target community (n = 112) via group discussions, interviews and drawings/stories of 'most significant change'. Households' handwashing/water facilities and toilets were observed. Analysis was informed by a model that highlights environmental, psychosocial and technological factors that shape hygiene behaviours across multiple levels, from the habitual to the structural (Dreibelbis et al. 2013). Findings indicate the intervention has supported development of new norms around hygiene behaviours. Key drivers of sustained hygiene behaviour were habit formation, emotional drivers (e.g. disgust, affiliation), and collective action and civic pride; key constraints included water scarcity and socio-economic disadvantage. Identifying and responding to the drivers and constraints of hygiene behaviour change in specific contexts is critical to sustained behaviour change and population health impact. Copyright © 2016 Elsevier Ltd. All rights reserved.

  5. Geothermobarometry in albite-garnet orthogneisses: A case study from the Gran Paradiso nappe (Western Alps)

    Science.gov (United States)

    Le Goff, Elisabeth; Ballèvre, Michel

    1990-11-01

    The aim of this paper is to estimate syntectonic P- T conditions within albite- and garnet-bearing orthogneisses. These rocks are generally characterized by the assemblage quartz + albite + biotite + phengite + CaFe-garnet + epidote + titanite. Garnet contains up to 55 mole per cent of grossular. K-feldspar is a relict magmatic phase. P- T conditions are estimated using several independent methods. First, it is shown that exchange reactions based on the FeMg partitioning between garnet and biotite or garnet and phengite cannot be used to estimate temperatures in these rocks, due to the high grossular content of garnet. Second, maximum and minimum pressures are constrained, respectively, by the occurrence of albite instead of jadeite + quartz and by the assemblage phengite + biotite + quartz. Third, phase equilibria in albite- and garnet-bearing metagranites are modelled in the system K 2OCaOFeOAl 2O 3SiO 2H 2O. Equilibrium curves are calculated for the observed phase compositions. Uncertainties in P- T estimates mainly result from the choice of appropriate non-ideal solution models for the garnet. An application is developed for granites from the Gran Paradiso nappe (Western Alps). These granites show an heterogeneous deformation of Alpine age expressed by mylonitic shear zones cutting across weakly deformed domains. Estimated P- T conditions for the synkinematic assemblages are 10-16 kbar at 550±50°C.

  6. Urban floods: a case study in the Savigliano area (North-Western Italy

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    C. Audisio

    2011-11-01

    Full Text Available Flood processes and effects are examined, concerning two rivers in an urbanized area in North-Western Italy (Piedmont – Cuneo Plain. In May 2008, some areas in Northern Italy were struck by intense and persistent rainfall. In the Cuneo province (Southern Piedmont, floodplain with some urban areas was inundated over ca. ten square kilometres, and the city of Savigliano (about 21 000 inhabitants was particularly hit by flood. A purposely-made historical research has evidenced approximately fifty flood events as having occurred since 1350 in the Savigliano area. Based upon historical data, both documents and maps, GIS (Geographical Information System technique and field surveys were used to quantitatively assess the growing urbanization of the city and to describe flood processes and effects over years. This work aims to describe the dynamic behaviour of the 2008 flood, also comparing it to past events, in particular those that occurred in 1896. It is emphasized how the knowledge of past events can be helpful in reducing urban flooding.

  7. Pemphigus in North-Western Yemen: A therapeutic study of 75 cases.

    Science.gov (United States)

    Khatri, Mishri Lal

    2016-01-01

    The incidence of pemphigus, though not documented, seems to be quite high in Yemen. There is no universal consensus on the treatment of this disease. The aim was to evaluate the efficacy and side effects of different therapeutic regimens used in patients of pemphigus in North-Western Yemen. Seventy-five Yemeni patients (39 males and 36 females) were included. Diagnosis was based on clinical features, histopathology and the Tzanck test. Results of treatment with these different therapeutic regimens were compared: (1) dexamethasone-cyclophosphamide pulse (DCP), (2) dexamethasone pulse with oral azathioprine, (3) oral prednisolone with azathioprine, (4) oral prednisolone with oral cyclophosphamide, and (5) prednisolone monotherapy. Pemphigus vulgaris (PV) was diagnosed in 46 patients, pemphigus foliaceus (PF) in 23, pemphigus vegetans (PVEG) in 5 and pemphigus herpetiformis (PH) in one. Among the 16 patients who received regular DCP therapy, 13 were in remission for 6 months to 11 years without medications (phase 4). Remission without pharmacotherapy could not be achieved with the other regimens and steroid-induced side-effects appeared to be more than with DCP. Immunofluorescence was not available to confirm the diagnosis of pemphigus. Randomization was not done. The DCP regimen seemed to be superior to the other regimens used.

  8. A Case Study of Nitrogen Saturation in Western U.S. Forests

    Directory of Open Access Journals (Sweden)

    Mark E. Fenn

    2001-01-01

    Full Text Available Virtually complete nitrification of the available ammonium in soil and nitrification activity in the forest floor are important factors predisposing forests in the San Bernardino Mountains of southern California to nitrogen (N saturation. As a result, inorganic N in the soil solution is dominated by nitrate. High nitrification rates also generate elevated nitric oxide (NO emissions from soil. High-base cation saturation of these soils means that soil calcium depletion or effects associated with soil acidification are not an immediate risk for forest health as has been postulated for mesic forests in the eastern U.S. Physiological disturbance (e.g., altered carbon [C] cycling, reduced fine root biomass, premature needle abscission of ozone-sensitive ponderosa pine trees exposed to high N deposition and high ozone levels appear to be the greater threat to forest sustainability. However, N deposition appears to offset the aboveground growth depression effects of ozone exposure. High nitrification activity reported for many western ecosystems suggests that with chronic N inputs these systems are prone to N saturation and hydrologic and gaseous losses of N. High runoff during the winter wet season in California forests under a Mediterranean climate may further predispose these watersheds to high nitrate leachate losses. After 4 years of N fertilization at a severely N saturated site in the San Bernardino Mountains, bole growth unexpectedly increased. Reduced C allocation below- ground at this site, presumably in response to ozone or N or both pollutants, may enhance the bole growth response to added N.

  9. How do pregnancy outcomes differ in teenage mothers? A Western Australian study.

    Science.gov (United States)

    Lewis, Lucy N; Hickey, Martha; Doherty, Dorota A; Skinner, S Rachel

    2009-05-18

    To determine whether teenage pregnancy and Indigenous status are associated with increased risk of adverse pregnancy outcomes. A cross-sectional descriptive analysis of nulliparous women with singleton pregnancies who delivered at the sole tertiary obstetric hospital in Western Australia between June 2004 and September 2006, using data obtained from computerised midwifery records. Maternal risk factors, pregnancy characteristics, and obstetric and perinatal outcomes for teenage and adult pregnancies. Of the 4896 births reviewed, 560 (11%) were to teenage mothers. Teenagers were more likely to be Indigenous and to experience maternal risk factors such as anaemia and smoking. Indigenous women were more likely than non-Indigenous women to be smokers, with young Indigenous teenagers (aged 12-16 years) being most likely to smoke (odds ratio [OR], 6.29; 95% CI, 3.99-9.92). Perinatal outcomes for teenage and adult births were similar, while adjustment for smoking and Indigenous status changed the observed association for the Indigenous population of preterm delivery teenagers (aged 17-18 years) were the group at highest risk of stillbirth (OR, 1.99; 95% CI, 1.03-3.76). These results improve our understanding of the obstetric and medical issues associated with teenage pregnancy and birth in WA and how we might tailor our approach to care. Indigenous teenagers need special attention, and there is significant scope for public health interventions around anaemia and smoking in this population.

  10. Reviving wood-pastures for biodiversity and people: A case study from western Estonia.

    Science.gov (United States)

    Roellig, Marlene; Sutcliffe, Laura M E; Sammul, Marek; von Wehrden, Henrik; Newig, Jens; Fischer, Joern

    2016-03-01

    Wood-pastures are associated with high cultural and biodiversity values in Europe. However, due to their relatively low productivity, large areas of wood-pastures have been lost over the last century. In some areas, incentive schemes have been developed to revive wood-pastures. We investigated the effects of one such scheme in western Estonia. We compared the structure of grazed wood-pastures (old and restored) to those of abandoned wood-pastures and ungrazed forest stands to explore the effects of management, and conducted interviews with 24 farmers to investigate their motivations to carry out the management. We found a positive influence of active management on the semi-open structure of wood-pastures. Financial support was vital for management, but personal values related to tradition also played an important role. The interviewees differed widely in their range of motivations, suggesting that other strategies in addition to financial incentives would further improve the management of wood-pastures in the region.

  11. PCB concentrations in plankton size classes, a temporal study in Marseille Bay, Western Mediterranean Sea.

    Science.gov (United States)

    Tiano, Marion; Tronczyński, Jacek; Harmelin-Vivien, Mireille; Tixier, Céline; Carlotti, François

    2014-12-15

    PCB levels in plankton were investigated in the Bay of Marseille, Western Mediterranean Sea, between September 2010 and October 2011. Concentrations of PCB congeners (CB 18, CB 52, CB 101, CB 118, CB 138, CB 153, CB 180) were determined in three plankton size-classes (60-200, 200-500 and 500-1000μm) together with different parameters: chlorophyll content, plankton dry-weight biomass, carbon and nitrogen stable isotope ratios and plankton-community structure. The ∑PCB7 concentrations ranged between 14.2 and 88.1ngg(-1)d.w., for all size classes and all sampling periods. The results do not show the biomass dilution effect and indicate moderate but significant biomagnification with plankton trophic position estimated by δ(15)N signatures. Equilibrium with water phase may notably contribute in controlling the PCB levels in the plankton. More generally, presented results imply that PCB accumulation in the plankton is an effect of abiotic and trophic complex interactions in the Bay of Marseille. Copyright © 2014 Elsevier Ltd. All rights reserved.

  12. [Comparative study of three Western models of deontological codes for dentists].

    Science.gov (United States)

    Macpherson Mayol, Ignacio; Roqué Sánchez, María Victoria; Gonzalvo-Cirac, Margarita; de Ribot, Eduard

    2013-01-01

    We performed a comparative analysis of the codes of ethics of three official organizations in Dentistry professional ethics: Code of Ethics for Dentists in the European Union, drawn up by the Council of European Dentists (CED); Código Español de Ética y Deontología Dental, published by the Consejo General de Colegios de Odontólogos y Estomatólogos de España (CGCOE); and Principles of Ethics and Code of Professional Conduct, of the American Dental Association (ADA). The analysis of the structure of the codes allows the discovery of different approaches governing professional ethics according to the ethical and legislative tradition from which they derive. While there are common elements inherent in Western culture, there are nuances in the grounds, the layout and wording of articles that allows to deduce the ethical foundations that underlie each code, and reflects the real problems encountered by dentists in the practice of their profession.

  13. Adherence to the Western Pattern Is Potentially an Unfavorable Indicator of Asthenozoospermia Risk: A Case-Control Study.

    Science.gov (United States)

    Eslamian, Ghazaleh; Amirjannati, Naser; Rashidkhani, Bahram; Sadeghi, Mohammad-Reza; Baghestani, Ahmad-Reza; Hekmatdoost, Azita

    2016-01-01

    The aim of this case-control study was to examine the relationship between dietary patterns and asthenozoospermia risk. In total, 107 incident asthenozoospermic men and 235 age-matched controls were interviewed through the infertility clinics in Tehran, Iran, from January 2012 to November 2013. Usual dietary intakes were collected using a semiquantitative food frequency questionnaire and semen quality data were analyzed according to the fifth edition of the World Health Organization (WHO) guidelines. Dietary patterns were derived using factor analysis. The first tertile served as the reference category for regression analyses. In principal component analysis, 2 dietary patterns emerged: a "prudent pattern" (leafy green vegetables, yellow vegetables, other vegetables, tomatoes, fish and other seafood, fruits and natural fruit juices, legumes, whole grains, poultry, tea and coffee, low-fat dairy products, and vegetable oils) and a "Western pattern" (organ meats, red and processed meats, sugar, soft drinks and confectionary, pasta, rice and refined grains, potatoes, french fries and fast foods, high-fat dairy products, hydrogenated fats, mayonnaise and fatty sauces, and snacks). After adjustment for potential confounders, participants in the highest tertile of the prudent pattern scores had 54% lower risk of asthenozoospermia compared to those in the lowest (p for trend: 0.003). Being in the highest tertile of the Western pattern was positively associated with asthenozoospermia risk (odds ratio [OR] = 2.86; 95% confidence interval [CI], 1.83-2.97). Our findings suggest that adherence to the Western pattern is potentially an unfavorable indicator of asthenozoospermia risk and a diet composed mainly of plant-based foods may be associated with a reduced risk.

  14. A random comparative study of terbinafine versus griseofulvin in patients with tinea capitis in Western China.

    Science.gov (United States)

    Deng, S; Hu, H; Abliz, P; Wan, Z; Wang, A; Cheng, W; Li, R

    2011-11-01

    To compare the efficacy and safety of terbinafine with griseofulvin in the treatment of tinea capitis in Western China. Children (2-14 years of age) with clinically diagnosed and potassium hydroxide microscopy-confirmed tinea capitis were randomized into three groups: group GRI4 received 4 weeks of griseofulvin; group TBF2 received 2 weeks of terbinafine; and Group TBF4 received 4 weeks of terbinafine. Clinical and mycological evaluations were done in 0, 2, 4, and 8 weeks and 1 year after therapy started. The isolated pathogenic fungi were evaluated for in vitro susceptibility by detecting the minimal inhibitory concentration (MIC) against terbinafine, griseofulvin, itraconazole, and ketoconazole. The clinical effectiveness rate of GRI4, TBF2, and TBF4 were 100% (95% CI-confidence interval: 82-100%), 96.3% (95% CI: 81-100%), and 100%(95% CI: 85-100%), respectively, at week 8 and 100% after 1 year for the 3 groups; clinical cure rates were 84.2%(95% CI: 77-99%), 85.2%(95% CI: 71-98%), and 78.3%(95% CI: 61-95%), respectively, at week 8 and 100% after 1 year for all agents; mycological cure rates were 100%(95% CI: 74-100%), 95.0%(95% CI: 74-100%), and 94.1%(95% CI: 50-93%) at week 8 and 100% after 1 year for the 3 groups. In vitro, all patient-derived cultures were sensitive to the four antifungal agents. Data from the clinical trial and in vitro antifungal activity indicated that terbinafine is efficacious and well tolerated in the treatment for Trichophyton infections (T. violaceum; Arthroderma vanbreuseghemii; and T. tonsurans) of the scalp, i.e., a 2- to 4-week course of terbinafine is as effective as a 4-week course of griseofulvin; in fact, a 2-week course of terbinafine is sufficient. Terbinafine is an effective alternative to griseofulvin against tinea capitis of Trichophyton infections.

  15. Pregnancy outcome in women with epilepsy in Western China: A prospective hospital based study.

    Science.gov (United States)

    He, Shixu; Zhu, Huili; Qiu, Xiangmiao; Zhu, Xi; Peng, Anjiao; Duan, Jianan; Chen, Lei

    2017-09-01

    Gaps exist in the diagnosis and treatment of women with epilepsy (WWE) between China and Euro-American countries. We aim to find out and share our experience of the multidisciplinary integrated treatment for WWE. We prospectively registered WWE who were diagnosed by both epileptologists and obstetrician in our green way system for the past 5years (2009-2015). Registration information include years of education, epilepsy history, seizure type and frequency, pregnancy and delivery complications, delivery mode, and Apgar score of newborn. All data were analyzed by SAS 9.3 version. We included 137 cases of maternal epilepsy (155 pregnancies with average maternal age of 26years old). 18 cases underwent epilepsy surgery before pregnancy. 103 pregnancies (66.45%) were cesarean section, 52 (33.55%) were natural childbirth, only 10 pregnancies have pregnancy complications, 2 have delivery complication, and 15 have seizures during delivery process. Most offspring were healthy when they were born (only 11 newborn got Apgar scoreartificial feeding, followed by 39 (16.77%) of mixed feeding and 26 of breastfeeding. Clinical features and perinatal outcome of Chinese WWE are similar to western WWE. For mode of delivery, even suggested by our epileptologists and obstetrician to deliver naturally, more patients selected cesarean section. Moreover, withdrawal of AEDs during pregnancy is common. Therefore, it is necessary to pay more attention to standard management of WWE and establish a more practical green way for WWE in China, to keep up with developed countries and improve the health level of birth in China. Copyright © 2017 Elsevier Inc. All rights reserved.

  16. Geohelminth Infections among Pregnant Women in Rural Western Kenya; a Cross-Sectional Study

    Science.gov (United States)

    van Eijk, Anna M.; Lindblade, Kim A.; Odhiambo, Frank; Peterson, Elizabeth; Rosen, Daniel H.; Karanja, Diana; Ayisi, John G.; Shi, Ya Ping; Adazu, Kubaje; Slutsker, Laurence

    2009-01-01

    Background Geohelminth infections are common in rural western Kenya, but risk factors and effects among pregnant women are not clear. Methodology During a community-based cross-sectional survey, pregnant women were interviewed and asked to provide a blood sample and a single fecal sample. Hemoglobin was measured and a blood slide examined for malaria. Geohelminth infections were identified using the concentration and Kato-Katz method. Results Among 390 participants who provided a stool sample, 76.2% were infected with at least one geohelminth: 52.3% with Ascaris lumbricoides, 39.5% with hookworm, and 29.0% with Trichuris trichiura. Infection with at least one geohelminth species was associated with the use of an unprotected water source (adjusted odds ratio [AOR] 1.8, 95% confidence interval [CI] 1.1–3.0) and the lack of treatment of drinking water (AOR 1.8, 95% CI 1.1–3.1). Geohelminth infections were not associated with clinical symptoms, or low body mass index. A hookworm infection was associated with a lower mid upper arm circumference (adjusted mean decrease 0.7 cm, 95% CI 0.3–1.2 cm). Hookworm infections with an egg count ≥1000/gram feces (11 women) were associated with lower hemoglobin (adjusted mean decrease 1.5 g/dl, 95% CI 0.3–2.7). Among gravidae 2 and 3, women with A. lumbricoides were less likely to have malaria parasitemia (OR 0.4, 95% CI 0.2–0.8) compared to women without A. lumbricoides, unlike other gravidity groups. Conclusion Geohelminth infections are common in this pregnant population; however, there were few observed detrimental effects. Routine provision of antihelminth treatment during an antenatal clinic visit is recommended, but in this area an evaluation of the impact on pregnancy, malaria, and birth outcome is useful. PMID:19172184

  17. Contributions to the Study of the Beginnings of the Orthodox Church in Western Rus’

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    Radosław Liwoch

    2014-11-01

    Full Text Available In 1882 T. Ziemięcki conducted in Pìdgìrcì (Lviv district, Ukraine a research excavation of two neighboring burial mounds which he called the Great Twin Barrows No. 1 and No. 2. In barrow No., he discovered on richly furnished double burial (probably of a married couple where he found two identical silver crosses. The crosses belong to the so called Scandinavian type (variant B.1 type 1.4.3 acc. J. Staecker. In barrow No. 2, he found a skeleton of a male buried with furnishings, including two crosses. The bigger one (lost was a bronze encolpion depicting the Crucifixion and the Virgin Orans, which can find analogies in the Danube countries. The smaller one is a silver St. Peter’s cross. In the mouths of the corpses found in both barrows there were tiny gold objects which could be interpreted as a local variation of the obolus of the dead custom. Both graves are a part of necropolis with druzhina burials. They date back to the end of the 10th century or the first half of the 11th century. The objects presented here are the earliest evidence to confirm the presence of a Christianized (Greek version of Christianity elite in the region of the Upper Bug and Upper Dniester, which is not burdened with chronological or interpretative reservation. The deceased were not members of the clergy but, nevertheless, representatives of the new official religion in a pagan Slavic population whose conversion had barely begun. It seems, therefore, that their presence in Pìdgìrcì can be considered the beginning of the Orthodox Church in the western part of the former Kievan Rus’.

  18. Miniaturized fluorescent RNA dot blot method for rapid quantitation of gene expression

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    Yadetie Fekadu

    2004-06-01

    Full Text Available Abstract Background RNA dot blot hybridization is a commonly used technique for gene expression assays. However, membrane based RNA dot/slot blot hybridization is time consuming, requires large amounts of RNA, and is less suited for parallel assays of more than one gene at a time. Here, we describe a glass-slide based miniaturized RNA dot blot (RNA array procedure for rapid and parallel gene expression analysis using fluorescently labeled probes. Results RNA arrays were prepared by simple manual spotting of RNA onto amino-silane coated microarray glass slides, and used for two-color fluorescent hybridization with specific probes labeled with Cy3 and 18S ribosomal RNA house-keeping gene probe labeled with Cy5 fluorescent dyes. After hybridization, arrays were scanned on a fluorescent microarray scanner and images analyzed using microarray image analysis software. We demonstrate that this method gives comparable results to Northern blot analysis, and enables high throughput quantification of transcripts from nanogram quantities of total RNA in hundreds of samples. Conclusion RNA array on glass slide and detection by fluorescently labeled probes can be used for rapid and parallel gene expression analysis. The method is particularly well suited for gene expression assays that involve quantitation of many transcripts in large numbers of samples.

  19. Hydrostratigraphy and hydrogeology of the western part of Maira area, Khyber Pakhtunkhwa, Pakistan: a case study by using electrical resistivity.

    Science.gov (United States)

    Farid, Asam; Jadoon, Khanzaib; Akhter, Gulraiz; Iqbal, Muhammad Asim

    2013-03-01

    Hydrostratigraphy and hydrogeology of the Maira vicinity is important for the characterization of aquifer system and developing numerical groundwater flow models to predict the future availability of the water resource. Conventionally, the aquifer parameters are obtained by the analysis of pumping tests data which provide limited spatial information and turn out to be costly and time consuming. Vertical electrical soundings and pump testing of boreholes were conducted to delineate the aquifer system at the western part of the Maira area, Khyber Pakhtun Khwa, Pakistan. Aquifer lithology in the eastern part of the study area is dominated by coarse sand and gravel whereas the western part is characterized by fine sand. An attempt has been made to estimate the hydraulic conductivity of the aquifer system by establishing a relationship between the pumping test results and vertical electrical soundings by using regression technique. The relationship is applied to the area along the resistivity profiles where boreholes are not drilled. Our findings show a good match between pumped hydraulic conductivity and estimated hydraulic conductivity. In case of sparse borehole data, regression technique is useful in estimating hydraulic properties for aquifers with varying lithology.

  20. The Development Needs of Newly Appointed Senior School Leaders in the Western Cape South Africa: A Case Study

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    Nelius Jansen van Vuuren

    2017-12-01

    Full Text Available The essential role that senior school leaders play in school leadership teams to ensure effective strategic leadership in schools has been the subject of intense discussion for many years. Crucial to this debate is the establishment of professional learning and leadership approaches for newly appointed senior school leaders. Recommendations for policy and practice highlight the importance of appropriate, multifaceted, developmental support initiatives for newly appointed school leaders. In many countries, including South Africa, a teaching qualification and, in most cases, extensive teaching experience is the only requirement for being appointed as a senior school leader in a school. This tends to suggest that no further professional development is required for newly appointed school leaders, the problem addressed in this paper. This paper reports on the main findings of the perceived development needs of newly appointed senior school leaders in the Western Cape, South Africa, and suggests that school leaders occupy a unique and specialist role in education, which requires relevant and specific preparation to support effective leadership. The respondents of this study report a lack of contextualised training and support before and after their appointment in their new roles creating unique development needs. This paper, therefore, employs a mixed-method approach to gather data to understand the perceived needs of twenty newly appointed senior school leaders in the Western Cape, South Africa.

  1. Modeling of a lot scale rainwater tank system in XP-SWMM: a case study in Western Sydney, Australia.

    Science.gov (United States)

    van der Sterren, Marlène; Rahman, Ataur; Ryan, Garry

    2014-08-01

    Lot scale rainwater tank system modeling is often used in sustainable urban storm water management, particularly to estimate the reduction in the storm water run-off and pollutant wash-off at the lot scale. These rainwater tank models often cannot be adequately calibrated and validated due to limited availability of observed rainwater tank quantity and quality data. This paper presents calibration and validation of a lot scale rainwater tank system model using XP-SWMM utilizing data collected from two rainwater tank systems located in Western Sydney, Australia. The modeling considers run-off peak and volume in and out of the rainwater tank system and also a number of water quality parameters (Total Phosphorus (TP), Total Nitrogen (TN) and Total Solids (TS)). It has been found that XP-SWMM can be used successfully to develop a lot scale rainwater system model within an acceptable error margin. It has been shown that TP and TS can be predicted more accurately than TN using the developed model. In addition, it was found that a significant reduction in storm water run-off discharge can be achieved as a result of the rainwater tank up to about one year average recurrence interval rainfall event. The model parameter set assembled in this study can be used for developing lot scale rainwater tank system models at other locations in the Western Sydney region and in other parts of Australia with necessary adjustments for the local site characteristics. Copyright © 2014 Elsevier Ltd. All rights reserved.

  2. Hydrostratigraphy and hydrogeology of the western part of Maira area, Khyber Pakhtunkhwa, Pakistan: A case study by using electrical resistivity

    KAUST Repository

    Farid, Asam M.

    2012-06-27

    Hydrostratigraphy and hydrogeology of the Maira vicinity is important for the characterization of aquifer system and developing numerical groundwater flow models to predict the future availability of the water resource. Conventionally, the aquifer parameters are obtained by the analysis of pumping tests data which provide limited spatial information and turn out to be costly and time consuming. Vertical electrical soundings and pump testing of boreholes were conducted to delineate the aquifer system at the western part of the Maira area, Khyber Pakhtun Khwa, Pakistan. Aquifer lithology in the eastern part of the study area is dominated by coarse sand and gravel whereas the western part is characterized by fine sand. An attempt has been made to estimate the hydraulic conductivity of the aquifer system by establishing a relationship between the pumping test results and vertical electrical soundings by using regression technique. The relationship is applied to the area along the resistivity profiles where boreholes are not drilled. Our findings show a good match between pumped hydraulic conductivity and estimated hydraulic conductivity. In case of sparse borehole data, regression technique is useful in estimating hydraulic properties for aquifers with varying lithology. © 2012 Springer Science+Business Media B.V.

  3. An ethnobotanical study of medicinal plants used by ethnic people in Parbat district of western Nepal.

    Science.gov (United States)

    Malla, Birendra; Gauchan, Dhurva P; Chhetri, Ran B

    2015-05-13

    Nepal, a mountainous country having diversified topographic and climatic conditions, gives rise to a wide range of flora and fauna. In villages ethnic people, including Magar and Majhi highly depend on medicinal plants for their primary healthcare. Due to overexploitation, encroachment in forest for arable land and destruction of natural habitat, potentially useful medicinal plants are going to be threatened in their natural environment. There is a strong need to enlist highly valuable medicinal plants and use traditional knowledge to protect, utilize and manage them in ex-situ and in-situ conservation. The present research investigates and documents traditional knowledge on medicinal plants utilization as natural medicine by two ethnic communities Magar and Majhi of Parbat district in Western Nepal. Ethnomedicinal data was collected during 2012-2013 by the following standard ethnobotanical methods. Data about medicinal uses of herbs, shrubs and trees were collected using semi-structured questionnaire, field observation, personal interview and group discussion with 334 (130 men and 204 women) pre-identified local informants, among which 13 male and 2 female were local healers. Statistical tool, informant consensus factors (F(IC)) and fidelity level (FL) values were used to analyze the importance of ethnomedicinal plants. A total of 132 ethnomedicinal plant species belonging to 99 genera and 67 families have been documented. These plants are used to treat various diseases and disorders grouped under 12 disease categories, with the highest number of species (61) being used for gastro-intestinal, parasitic and hepatobiliary disorders (FIC=0.78%), followed by blood and lymphatic system (F(IC)=0.76%) category. The highest fidelity level (FL) values recorded in Paris polyphylla (FL=96.0%) followed by Bergenia ciliata (FL=95.0%) confirms that these plants are the best plant species with medicinal properties. The two ethnic communities, Magar and Majhi, in Parbat district

  4. A Case Study in Archaeoseismology: the Collapses of the Temples at Selinunte (South-Western Sicily)

    Science.gov (United States)

    Guidoboni, E.

    2001-12-01

    This work presents the results of a research that was carried out to further our knowledge of the active faults in south-western Sicily. The only seismic event of particular significance that is known to have occurred is that of 15 January 1968 (Io= XMCS, Me 6.5) in the Belice valley. The archaeological park of Selinunte, the largest in the Mediterranean area, with its great temples and evidence of spectacular collapses, has been taken as a source of information capable of analysis by means of the methodological approach of archaeoseismology. The identification of the seismic indicators at Selinunte necessitated a detailed analysis of both old and new archaeological evidence, together with a critical re-examination of all the archaeological literature and existing documents dating from to the eighteenth century, together with travelers' accounts of that time. The history of the archaeological deposits, spoliation, and excavations has been reconstructed. These data are reinterpreted in the light of both the new discoveries of the most recent research, and of a number of methodological criteria already used in previous works on archaeoseismology. This long and complex analysis was carried out in 1998-99 with Anna Muggia, Clemente Marconi and Enzo Boschi in the research programme of the Istituto Nazionale di Geofisica e Vulcanologia. It resulted in the identification of two seismic events that struck Selinunte and led to the collapse of the temples. The chronology of the earthquakes can be dated, for the first, to a period between the fourth and third century BC; for the second, to a period between the sixth and the thirteen century AD. Although the time span proposed, particularly for the second earthquake, is very broad, it does not mean that this earthquake is in anyway hypothetical from a geophysical point of view. Its traces are clear, but the long periods when the site was abandoned mitigate against the fixing of a precise date for the event. In order to reach

  5. Crustal structure of western Hispaniola (Haiti) from a teleseismic receiver function study

    Science.gov (United States)

    Corbeau, J.; Rolandone, F.; Leroy, S.; Guerrier, K.; Keir, D.; Stuart, G.; Clouard, V.; Gallacher, R.; Ulysse, S.; Boisson, D.; Bien-aimé Momplaisir, R.; Saint Preux, F.; Prépetit, C.; Saurel, J.-M.; Mercier de Lépinay, B.; Meyer, B.

    2017-07-01

    Haiti, located at the northern Caribbean plate boundary, records a geological history of terrane accretion from Cretaceous island arc formations to the Eocene to Recent oblique collision with the Bahamas platform. Little is presently known about the underlying crustal structure of the island. We analyze P-waveforms arriving at 27 temporary broadband seismic stations deployed over a distance of 200 km across the major terrane boundaries in Haiti to determine the crustal structure of western Hispaniola. We compute teleseismic receiver functions using the Extended-Time Multi-Taper method and determine crustal thickness and bulk composition (Vp/Vs) using the H-k stacking method. Three distinctive and fault-bounded crustal domains, defined by their characteristic Moho depth distributions and bulk crustal Vp/Vs, are imaged across Haiti. We relate these domains to three crustal terranes that have been accreted along the plate boundary during the northeastwards displacement of the Caribbean plate and are presently being deformed in a localized fold and thrust belt. In the northern domain, made up of volcanic arc facies, the crust has a thickness of 23 km and Vp/Vs of 1.75 ± 0.1 typical of average continental crust. The crust in the southern domain is part of the Caribbean Large Igneous Province (Caribbean LIP), and is 22 km thick with Vp/Vs of 1.80 ± 0.03 consistent with plume-related rocks of late Cretaceous age. Significantly thicker, the crust in central Haiti has values of Moho depths averaging 41 km and with Vp/Vs of 1.80 ± 0.05. We propose that the central domain is likely constructed of an island arc upper crust with fragments of dense material originating from mafic lavas or LIP material. We produce a crustal profile along a N-S transect across Haiti accounting for the surface geology, shallow structural history, and new seismological constraints provided by variations of crustal thickness and bulk composition.

  6. A Case Study of a Methodological Approach to Cocreating Perinatal Health Knowledge Between Western and Indigenous Communities

    Directory of Open Access Journals (Sweden)

    Sujane Kandasamy

    2017-03-01

    Full Text Available This article describes the methods taken to create an understanding of the perinatal health beliefs of elder Indigenous women of the Six Nations of the Grand River in Ontario, Canada. Our study paired constructivist grounded theory data collection and analysis methods with an Indigenous epistemological framework. We aimed to create knowledge that was specific to an Indigenous context, which was useful and resonant with both Indigenous and Western readers. The multidisciplinary research team included Indigenous and non-Indigenous members and worked with a common appreciation for multiple knowledge sources. We offer an account of our process and methodological principles to serve as an illustrative case study of bringing together diverse approaches when working with Indigenous communities.

  7. A comparative study on genetic and environmental influences on metabolic phenotypes in Eastern (Chinese) and Western (Danish) populations

    DEFF Research Database (Denmark)

    Li, Shuxia

    2015-01-01

    and conducted the comparative study between the two populations, Danish data represent western and Chinese data represent eastern populations. Results from this PhD project suggested that there are disparity patterns of genetic and environmental regulations of some important metabolic phenotypes between Danish......During the last decades metabolic disorders such as high blood pressure, impaired blood glycose, atherosclerotic lipid abnormalities, overweight/obesity and elevated blood pressure are among the leading risks for mortality and morbidity worldwide. Those risks are mostly responsible for raising...... the risk of clinic diseases e.g. diabetes, atherosclerosis, stroke and cardiovascular disease. Metabolic phenotypes, similar to most complex traits, can be influenced by both genetic and environmental factors as well as their interplay. Many family and twin studies have demonstrated both genetic...

  8. Study of water mixing in the coastal waters of the western Taiwan Strait based on radium isotopes.

    Science.gov (United States)

    Men, Wu; Jiang, Yuwu; Liu, Guangshan; Wang, Fenfen; Zhang, Yusheng

    2016-02-01

    Radium is considered to be a useful tracer for studying the physical processes of seawater. In this work, three naturally occurring radium isotopes, (224)Raex, (226)Ra and (228)Ra, were measured in the coastal zone of the western Taiwan Strait during the summer seasons. Based on the distributions of the three radium isotopes and the salinity, we conclude that the water mixing pattern in the study area in summer consists of diluted water flowing from the Jiulong River to the open sea towards the east and southeast, and open sea seawater flowing inward from south to north. The submarine ground water discharges in the estuarine region, as suggested by the radium and salinity data. The residence times of the Jiulong River estuary, ranging from 7 to 49 d, were estimated using the radium isotope pairs (224)Raex and (226)Ra. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Characterization of an atypical lipoprotein-binding protein in human aortic media membranes by ligand blotting.

    Science.gov (United States)

    Kuzmenko, Y S; Bochkov, V N; Philippova, M P; Tkachuk, V A; Resink, T J

    1994-01-01

    By use of ligand-blotting techniques, this study investigated lipoprotein-binding proteins in human aortic smooth muscle. PAGE was performed under non-reducing conditions, and, using low-density lipoprotein (LDL) as ligand, with rabbit anti-apolipoprotein (apo) B and 125I-labelled goat anti-rabbit IgG as primary and secondary antibodies respectively, we demonstrate that membranes from human aortic media (and cultured human smooth-muscle cells) contain a major lipoprotein-binding protein with an apparent molecular mass of 105 kDa. Anionized preparations (carbamoyl- and acetyl-) of LDL, which did not displace 125I-LDL bound to the apo B,E receptor of cultured fibroblasts, were also recognized as ligands for the 105 kDa protein in aortic media membranes. LDL binding to 105 kDa protein was decreased in the presence of high density lipoprotein (HDL), although more than 100-fold molar excess of HDL was required to achieve 50% displacement of bound LDL. The LDL-binding activity of 105 kDa protein was inhibited by EDTA, and was also significantly decreased when samples were reduced by beta-mercaptoethanol before electrophoresis. Monoclonal antibodies against apo B,E receptor reacted with partially purified bovine adrenal apo B,E receptor, but not with 105 kDa protein of human aortic media membranes. The spectrum of properties of this vascular smooth-muscle lipoprotein-binding protein binding are clearly distinct from those of other previously characterized lipoprotein-binding molecules. Images Figure 1 Figure 2 Figure 4 Figure 5 Figure 6 Figure 8 PMID:7945254

  10. 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.

  11. Molecular studies of BKCa channels in intracranial arteries: presence and localization

    DEFF Research Database (Denmark)

    Johansson, Helle Wulf; Hay-Schmidt, Anders; Poulsen, Asser Nyander

    2008-01-01

    of the BK(Ca) channel in rat basilar, middle cerebral, and middle meningeal arteries by reverse transcription polymerase chain reaction (RT-PCR), quantitative real-time PCR, and Western blotting. Distribution patterns were investigated using in situ hybridization and immunofluorescence studies. RT......-PCR and quantitative real-time PCR detected the expression of the BK(Ca) channel mRNA transcript in rat basilar, middle cerebral, and middle meningeal arteries, with the transcript being expressed more abundantly in rat basilar arteries than in middle cerebral and middle meningeal arteries. Western blotting detected...

  12. Molecular studies of BKCa channels in intracranial arteries

    DEFF Research Database (Denmark)

    Wulf, Helle; Hay-Schmidt, Anders; Poulsen, Asser Nyander

    2008-01-01

    expression of the BK(Ca) channel in rat basilar, middle cerebral, and middle meningeal arteries by reverse transcription polymerase chain reaction (RT-PCR), quantitative real-time PCR, and Western blotting. Distribution patterns were investigated using in situ hybridization and immunofluorescence studies. RT......-PCR and quantitative real-time PCR detected the expression of the BK(Ca) channel mRNA transcript in rat basilar, middle cerebral, and middle meningeal arteries, with the transcript being expressed more abundantly in rat basilar arteries than in middle cerebral and middle meningeal arteries. Western blotting detected...

  13. Intangible Capital, Innovation and Export-Led Growth: Empirical Comparative Study of Slovenia and the Western Balkans

    Directory of Open Access Journals (Sweden)

    Marija Drenkovska

    2015-03-01

    Full Text Available In the face of progressing globalisation and liberalisation of the markets, innovation is the minimum necessary requirement for companies and countries to be globally competitive, and knowledge is the key input. In a comparative study we investigate the intellectual capital of a sample of firms from the Western Balkans and Slovenia, and analyse the link between intellectual capital, innovation, and export volume. Using unique survey data sets for these countries, we propose a structural model to examine our hypotheses. The results suggest that possessing intellectual capital does not suffice for firms’ global competitiveness and that higher presence on global markets may offer exposure to more advanced knowledge that firms cannot obtain in their domestic markets.

  14. A Case-Study of the Resources and Functioning of Two Research Ethics Committees in Western India.

    Science.gov (United States)

    Chenneville, Tiffany; Menezes, Lynette; Kosambiya, Jayendrakumar; Baxi, Rajendra

    2016-12-01

    Assessing the resources and functioning of research ethics committees (RECs) in low-resource settings poses many challenges. We conducted a case study of two medical college RECs (A and B) in Western India utilizing the Research Ethics Committee Quality Assurance Self-Assessment Tool (RECQASAT) as well as in-depth interviews with representative members to evaluate REC effectiveness. REC A and B obtained 62% and 67% of allowable points on the RECQASAT. These scores together with findings from the in-depth interviews suggest the need for significant improvement in REC effectiveness particularly in the areas of membership and educational training, organizational aspects, recording minutes, communicating decisions, and REC resources. Developing evidence-based best practices and strengthening infrastructure are essential to enhancing REC efficacy in low-resource countries.

  15. Contributions to the phytosociological study of Sessile and Turkey oak forests in the Orastie area (Central-Western Romania

    Directory of Open Access Journals (Sweden)

    Petru BURESCU

    2012-11-01

    Full Text Available In the current paper we present a phytosociological study of the phytocoenoses of the association Quercetum petraeae-cerris Soó 1963 (Syn.: Quercetum polycarpae-cerris G. Popescu 1988 identified in the Orăştie river basin, situated in the central-western part of Romania.The characterisation of the association under analysis as well as the presentation of the phytosociological table have been done by selecting the most representative relevés performed in the Turkey and Sessile oak forests of the lower and central parts of the Orastie river basin (Culoarul Orăştiei.This paper aims at analysing the phytocoenoses of the association Quercetum petraeae-cerris Soó 1963 in terms of physiognomy and floristic composition, life forms spectrum, floristic elements, and ecological indices.

  16. Assessment of the Affective Evaluation Competencies of Social Studies Teachers in Secondary Schools in Western Nigeria

    Science.gov (United States)

    Omare, C. Otote; Iyamu, Ede O. S.

    2006-01-01

    The nature and objectives of Social Studies in Nigerian Secondary schools indicate the affective orientation of the subject. Studies abound on the dominance of cognitive orientation to the teaching and evaluation of the subject in the schools, an indication that the curriculum is poorly implemented. This study assessed the affective evaluation…

  17. The Response of Grain Potential Productivity to Land Use Change: A Case Study in Western Jilin, China

    Directory of Open Access Journals (Sweden)

    Fei Li

    2015-11-01

    Full Text Available The impact of land use change on grain potential productivity is one of the most important topics in the research of land use/cover change and its effects. Western Jilin, located on the edge of an ecotone in northern China, and its land use have changed dramatically in recent decades, with significant impact on grain potential productivity. This study evaluated the grain potential productivity in different conditions and analyzed the response to land use change based on land use data, meteorological data and statistical data by using the Global Agro-ecological Zone model. Results showed that (1 grain potential productivity of Western Jilin in 2010 was 19.12 million tons, an increase of 34.8% over 1975 because of changes in land use and in climate; (2 due to land use change, grain potential productivity in the study area increased between 1975 and 2000, however, it decreased between 2000 and 2010; (3 conversion in type of land use and an increase in irrigation percentage caused grain potential productivity to increase by 0.70 million tons and 3.13 million tons respectively between 1975 and 2000; between 2000 and 2010, grain potential productivity had an increase of 0.17 million tons due to the increase in farmland area, but it decreased by 1.88 million tons because the irrigation percentage declined from 36.6% to 24.7%. Therefore, increasing investment in agriculture, improving land quality and increasing the conversion rate of grain potential productivity to actual production would be a better choice for ensuring national food security and achieving sustainable land use.

  18. Complicated secondary textures in zircon record evolution of the host granitic rocks: Studies from Western Tauern Window and Ötztal-Stubai Crystalline Complex (Eastern Alps, Western Austria)

    Science.gov (United States)

    Kovaleva, Elizaveta; Harlov, Daniel; Klötzli, Urs

    2017-07-01

    Samples of metamorphosed and deformed granitic rocks were collected from two Alpine complexes with well-constrained metamorphic history: Western Tauern Window and Ötztal-Stubai Crystalline Complex. Zircon grains from these samples were investigated in situ by a combination of scanning electron microscope techniques, cathodoluminescence (CL) imaging and Raman spectroscopy. The aims were: to describe and interpret complicated secondary textures and microstructures in zircon; based on cross-cutting relationships between secondary microstructures, reconstruct the sequence of processes, affecting zircon crystals; link the evolution of zircon with the history of the host rocks. The results indicate that zircon in the sampled granitic rocks forms growth twins and multi-grain aggregates, which are unusual for this mineral. Moreover, various secondary textures have been found in the sampled zircon, often cross-cutting each other in a single crystal. These include: distorted oscillatory CL zoning with inner zones forming inward-penetrating, CL-bright embayments, which are the evidence of dry recrystallization via annealing/lattice recovery; CL mosaicism with no preservation of growth zoning, but abundant nano- and micro-scale pores and mineral inclusions, which are the evidence of recrystallization by coupled dissolution-reprecipitation and/or leaching; embayed zircon boundaries filled with apatite, monazite, epidote and mylonitic matrix, indicating mineral-fluid reactions resulting in zircon dissolution and fragmentation; overgrowth CL-dark rims, which contain nano-pores and point to transport and precipitation of dissolved zircon matter. We conclude that zircon in our meta-granites is sensitive to metamorphism/deformation events, and was reactive with metamorphic fluids. Additionally, we have found evidence of crystal-plastic deformation in the form of low angle boundaries and bent grain tips, which is a result of shearing and ductile deformation of the host rock. We

  19. The importance of forest islands for invertebrate biodiversity: a case study in Western Poland

    National Research Council Canada - National Science Library

    Manole, Traian; Banaszak, Józef; Ratyńska, Halina; Ionescu-Mălăncuş, Irina; Petrescu, Eugenia; Mărgărit, Gabriela

    2011-01-01

    Ecological landscape studies are carried out concerning the ecosystem biocenosis restore and conservation and to define the ecological terms like "ecosystem services" which have increasingly caught...

  20. 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.

  1. Convenient analysis of protein modification by chemical blotting with fluorogenic “click” reagents

    Science.gov (United States)

    Ohata, Jun; Farrukh, Vohidov; Ball, Zachary T.

    2015-01-01

    Direct visualization of bioorthogonal alkyne or azide handles using fluorogenic azide-alkyne cycloaddition conducted on the surface of a blot membrane. The method eliminates the need for separation steps to remove excess small molecule reagents before attachment of antigen molecules or other visualization handles, and is especially useful for the analysis of peptides and small proteins. A variety of potential fluorogenic reagents are assessed, and sensitivity (<0.1 picomole) similar to current commercially available fluorescence imaging methods is possible. PMID:26325302

  2. Multi-type maltreatment in childhood and psychological adjustment in adolescence: questionnaire study among adolescents in Western Herzegovina Canton.

    Science.gov (United States)

    Sesar, Kristina; Zivcić-Bećirević, Ivanka; Sesar, Damir

    2008-04-01

    To determine the prevalence and intercorrelation of different forms of childhood maltreatment and psychological problems in adolescents in Western Herzegovina Canton. A questionnaire study was conducted in March 2003 on a convenient sample of 458 third-grade high-school students (39% boys) aged between 15 and 20 (median age, 17). Data were collected using a sociodemographic questionnaire, Family Adaptability and Cohesion Evaluation Scales III, Child Maltreatment Questionnaire, Youth Self-Report, and Rosenberg Self-Esteem Scale. Sociodemographic and family characteristics and exposure to maltreatment were analyzed as possible predictors of exposure to a particular type of abuse and subsequent psychological adjustment problems. Out of 458 students, 77% were emotionally abused, 52% physically abused, 30% neglected, 20% witnessed family violence, and 13% of girls and 21% of boys were sexually abused before the age of 14. Significant association between the maltreatment by a mother, father, and other adults were found for emotional and physical abuse and for neglect and witnessing family violence (r=0.413-0.541, Pstudents were exposed to multi-type maltreatment in childhood. Family characteristics and maltreatment scores significantly predicted anxiety/depression (R=0.456, R(2)=0.076), withdrawal (R=0.389, R(2)=0.049), somatic complaints (R=0.437, R(2)=0.059), social problems (R=0.417, R(2)=0.063), attention deficit and hyperactivity disorder (R=0.368, R(2)=0.045), rule-breaking behavior (R=0.393, R(2)=0.045), aggression (R=0.437, R(2)=0.078) (Pstudents in Western Herzegovina Canton were exposed to multi-type maltreatment in childhood, regardless of the war experience. Emotional and physical abuse were most frequently combined forms of maltreatment. Sociodemographic and family characteristics and exposure to some forms of abuse were significant predictors of exposure to other forms of abuse. Exposure to maltreatment in childhood predicted difficulties in psychological

  3. Utility of pediatric cariogenicity index among preschoolers of the western part of Maharashtra: A cross-sectional study

    Directory of Open Access Journals (Sweden)

    Siddhi Pancholi

    2015-01-01

    Full Text Available Introduction: Early childhood caries (ECC is a multifactorial infectious disease occurring on one or more decayed, missing or filled tooth surfaces in a child from birth to 6 years of age. ECC results from the interaction of behavioral and biological factors including fermentable carbohydrate, plaque, and dietary patterns. Diet is a major modifiable risk factor in the initiation of ECC. Aim: The aim was to assess the utility of pediatric cariogenicity index among the Indian population. Materials and Methods: A cross-sectional study was conducted with sample size of 400 preschool children of Satara district, western Maharashtra. Data regarding the diet were obtained from caregivers and cariogenicity indices were used to score dietary data using a food frequency questionnaire. The 24 h dietary recall was taken. Preschool children were subjected to type 3 dental examinations. Solid and liquid cariogenicity scores were assessed by Statistical analysis - SPSS v 17.0. Results: The mean deft was 1.44 (±1.20 for the subjects. Similarly, according to the calculation using 24 h recall, mean solid cariogenicity score was 2.3 (±0.9 and mean liquid cariogenicity score 4.37 (±1.5. The correlation between deft and the solid cariogenicity score was not significant (r = −0.074. The correlation between deft and liquid cariogenicity scores was significant and positive (r = 0.671. Conclusions: There was definite positive correlation of liquid cariogenic food intake with the ECC experience among the preschool children of western part of Maharashtra.

  4. Association between dietary protein intake and the risk of hypertension: a cross-sectional study from rural western China.

    Science.gov (United States)

    Liu, Ruru; Dang, Shaonong; Yan, Hong; Wang, Duolao; Zhao, Yaling; Li, Qiang; Liu, Xiaoning

    2013-11-01

    Evidence for an association between dietary proteins and the risk of hypertension in rural Chinese adults, whose diets are protein-poor and unbalanced, is limited. The aim of this study was to investigate the effect of dietary proteins on hypertension among adults of rural western China. A cross-sectional survey was conducted in Hanzhong, China, and 2241 residents were included in the analysis. Logistic regression models were used to examine whether dietary proteins (total, plant and animal protein) were independently correlated with hypertension. The total protein intake was inadequate (51.7 g per day for male and 40.1 g per day for female), accounting for 56-71% of the Chinese recommended nutrient intakes or adequate intakes. Nearly 80% of protein intake derived from plants, especially grains, and the proportion derived from plants was higher in females than in males. The daily animal protein intake was 12.1 g for males and 8.3 g for females. For females, no significant association was found between hypertension and total protein or plant protein intake. However, animal protein intake was significantly and negatively associated with the risk of hypertension after controlling for demographic characteristics, lifestyle factors, body mass index and other dietary nutrients. In addition, the odds ratio for the upper quartile compared with the bottom quartile was 0.64 (95% confidence interval, CI: 0.43-0.95, P for trend intake in females. For males, we did not observe a significant association between dietary proteins and hypertension or blood pressure. In conclusion, for the western rural Chinese population, especially women, whose dietary protein intake is low and largely derived from plants, the intake of animal protein may be related to a decreased risk of hypertension.

  5. 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.