WorldWideScience

Sample records for western blot cambridge

  1. 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...

  2. 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.

  3. 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.

  4. 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.

  5. 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.

  6. Single cell-resolution western blotting.

    Science.gov (United States)

    Kang, Chi-Chih; Yamauchi, Kevin A; Vlassakis, Julea; Sinkala, Elly; Duncombe, Todd A; Herr, Amy E

    2016-08-01

    This protocol describes how to perform western blotting on individual cells to measure cell-to-cell variation in protein expression levels and protein state. Like conventional western blotting, single-cell western blotting (scWB) is particularly useful for protein targets that lack selective antibodies (e.g., isoforms) and in cases in which background signal from intact cells is confounding. scWB is performed on a microdevice that comprises an array of microwells molded in a thin layer of a polyacrylamide gel (PAG). The gel layer functions as both a molecular sieving matrix during PAGE and a blotting scaffold during immunoprobing. scWB involves five main stages: (i) gravity settling of cells into microwells; (ii) chemical lysis of cells in each microwell; (iii) PAGE of each single-cell lysate; (iv) exposure of the gel to UV light to blot (immobilize) proteins to the gel matrix; and (v) in-gel immunoprobing of immobilized proteins. Multiplexing can be achieved by probing with antibody cocktails and using antibody stripping/reprobing techniques, enabling detection of 10+ proteins in each cell. We also describe microdevice fabrication for both uniform and pore-gradient microgels. To extend in-gel immunoprobing to gels of small pore size, we describe an optional gel de-cross-linking protocol for more effective introduction of antibodies into the gel layer. Once the microdevice has been fabricated, the assay can be completed in 4-6 h by microfluidic novices and it generates high-selectivity, multiplexed data from single cells. The technique is relevant when direct measurement of proteins in single cells is needed, with applications spanning the fundamental biosciences to applied biomedicine.

  7. SDS -PAGE and Western Blotting Techniques.

    Science.gov (United States)

    Blancher, C; Jones, A

    2001-01-01

    The goal of Western blotting, or more correctly, immunoblotting, is to identify with a specific antibody a particular antigen within a complex mixture of proteins that has been fractionated in a polyacrylamide gel and immobilized onto a membrane. Immunoblotting can be used to determine a number of important characteristics of protein antigens-the presence and quantity of an antigen, the relative molecular weight of the polypeptide chain, and the efficiency of extraction of the antigen.Immunoblotting occurs in six stages: (1) extraction and quantification of protein samples; (2) resolution of the protein sample in sodium dodecyl sulfatepolyacrylamide denaturing gel electrophoresis (SDS-PAGE); (3) transfer of the separated polypeptides to a membrane support; (4) blocking nonspecific binding sites on the membrane; (5) addition of antibodies; and (6) detection.Sample preparation is important for obtaining accurate separation of the proteins on the basis of molecular weight. Depending on whether an antigen is primarily extracellular, cytoplasmic, or membrane-associated different procedures might be required to prepare the sample initially. Although there are exceptions, many soluble nuclear and cytoplasmic proteins can be solubilized by lysis buffers that contain the nonionic detergent Nonidet P-40 (NP-40) and either no salt at all or relatively high concentrations of salt (e.g., 0.5 M NaCl). However, the efficiency of extraction is often greatly affected by pH of the buffer and the presence or absence of chelating agents such EDTA.

  8. Selected Reaction Monitoring (SRM) frente a Western Blot

    OpenAIRE

    Heredia Ponce, Zahira Maria; Urbano Gámez, Jose Alberto

    2016-01-01

    We propose the SRM technology as a complementary method to the Western Blot for the detection and quantification of proteins in a sample. The technique Western Blot has its own limitations: i) only a protein-of-choice is detected, ignoring any non-relevant proteins, ii) the sensitivity of the technique depends on the specificity of the antibody and iii) Western Blot is expensive and time-consuming. The advantages of SRM with respect Western Blot are remarkable: i) you can detect up to h...

  9. 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.

  10. 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.

  11. The Design of a Quantitative Western Blot Experiment

    Directory of Open Access Journals (Sweden)

    Sean C. Taylor

    2014-01-01

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

  12. 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.

  13. 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.

  14. 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.

  15. 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

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

    NARCIS (Netherlands)

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

    2015-01-01

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

  17. 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.

  18. 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 disadvanta

  19. 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.

  20. 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.

  1. 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.

  2. 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.

  3. 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.

  4. 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.

  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. 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.

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

    Directory of Open Access Journals (Sweden)

    Brad L. Ericson

    2016-01-01

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

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

    Science.gov (United States)

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

    2006-01-01

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

  9. 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.

  10. 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.

  11. 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 A

    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....

  12. 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…

  13. 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.

  14. 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.

  15. 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

  16. 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).

  17. 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 antibo

  18. Use of Nonradioactive Detection Method for North- and South-Western Blot.

    Science.gov (United States)

    Franke, Claudia; Gräfe, Daniel; Bartsch, Holger; Bachmann, Michael P

    2015-01-01

    Many proteins bind to nucleic acids. For the first characterization of novel proteins, a fast and simple technique for testing their nucleic acid binding capabilities is desirable. Here we describe the use of a North-western and South-western blot protocol for the evaluation of the DNA and RNA binding abilities of a novel putative methyl transferase HSPC133 (METTL5).

  19. Evaluation of an indigenous western blot kit for human immunodeficiency virus

    Directory of Open Access Journals (Sweden)

    Lakshmi V

    2002-01-01

    Full Text Available PURPOSE: The Western Blot test is considered a gold standard test for the confirmation of an ELISA and/or rapid assay screened reactive sample in the diagnosis of HIV infection, especially in the low risk population. In this study, an indigenously developed HIV W. Blot kit (J.Mitra & Co., New Delhi, India was compared for its performance characteristics with a widely used Western Blot kit, HIV Blot 2.2 (Genelabs, Singapore. Antigens of both HIV-1 and the indicator antigen gp36 of HIV-2 are included in the strips. METHODS: A panel of 150 clinical serum samples was used in the evaluation. All the sera were tested simultaneously by both the kits. RESULTS: The HIV W. Blot kit had high performance characteristics (100% sensitivity and 100% specificity, like the HIV Blot 2.2. The test procedure was easy to perform. There was clear delineation of the bands. CONCLUSIONS: The interpretation of the results on the HIV W. Blot was less prone to subjective errors. The test gave positive bands at even very high serum dilutions in the test kit. This fact indicates that HIV W. Blot probably has a potential application in early phases of infection, when the antibody concentrations are still very low.

  20. 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.

  1. Quantum dot-based western blot for sensitive detection of pig serum antibody to actinobacillus pleuropneumoniae

    Science.gov (United States)

    Cişmileanu, Ana; Sima, Cornelia; Grigoriu, Constantin

    2007-08-01

    A quantum dot - immunoglobulin conjugate specific for pig IgG, was obtained by carbodiimide chemistry. We used a Western blot technique for detecting specific antibodies against Actinobacillus pleuropneumoniae (A. pp), which cause porcine pleuropneumonia. The antigen used in this technique was Apx haemolysin which is an important virulence factor of A. pp and it induces protective immunity in vaccined pigs. The detection on Western blot membrane was possible at 1/50 dilution of quantum dot conjugate at a dilution of pig serum till 1/6400. The results for pig serum demonstrated a higher sensitivity of QD-based Western blot technique for the presence of antibodies specific for Apx haemolysin in comparison with similar classical techniques (with coloured substrate for enzyme present in secondary antibody conjugate).

  2. 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.

  3. 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...

  4. 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...

  5. 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...

  6. 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…

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

    Science.gov (United States)

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

    2016-01-01

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

  8. 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…

  9. 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…

  10. Western blotting as a method for studying cell-biomaterial interactions : The role of protein collection

    NARCIS (Netherlands)

    van Kooten, TG; Klein, CL; Kirkpatrick, CJ

    2001-01-01

    Research of cell-biomaterial interactions is building on knowledge and methods available in cell and molecular biology. Western blotting is one of the options to characterize protein expression in cell populations. Method transfer to biomaterial model systems is not trivial because of the structure

  11. 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.

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

    Science.gov (United States)

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

    2016-03-01

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

  13. 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.

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

    OpenAIRE

    Son, Eui-Sun; Kim, Tong Soo; Nam, Ho-Woo

    2001-01-01

    Western blot analysis was performed to diagnose vivax malaria using stage-specific recombinant antigens. Genomic DNA from the whole blood of a malaria patient was used as templates to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), merozoite surface protein (MSP-1), apical merozoite antigen (AMA-1), serine repeat antigen (SERA), and exported antigen (EXP-1) of Plasmodium vivax. Each amplified DNA fragment was inserted into a pGEX-4T plasmid to induce ...

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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

  20. 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.

  1. 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.

  2. 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.

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

    DEFF Research Database (Denmark)

    Uhlén, Mathias

    2017-01-01

    applications and that this influences the epitopes exposed on the target protein, which might have profound consequences for the ability of a given antibody to bind specifically to its target. As an example, proteins that are analyzed by immunohistochemistry (IHC) are normally first cross-linked with formalin....... In conclusion, western blot and protein array analyses can indeed be useful tools when selecting specific antibodies for other applications. The use of these methods is encouraged both for antibody providers and users, and antibodies with signs of cross-reactivity in these applications should be treated...

  4. 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.

  5. 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

  6. 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.

  7. 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.

  8. 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.

  9. Analysis of sperm membrane antigens relevant to antisperm antibody using Western blot

    Institute of Scientific and Technical Information of China (English)

    WangHF

    2002-01-01

    Objective:To identify the sperm membrane antigens associated with antisperm antibody.Methods:The antisperm antibody in serum was tested by ELISA.Antisperm antibody positive sera from 18 infertile men and 15 infertile women were used.The molecular weight(MW) of sperm membrane antigens associated with the antisperm antibody was analyzed with antisperm antibody positive serum using Western blot.Results:Eight kinds of MW of sperm membrane antigens were identified.The ratio of identification on the 78 KD(60.7%),60KD(71.4%),51KD(14.9%) and 23KD(14.29%)sperm antigen was higher than other.Conclusion:sperm membrane antigens with MW of 78KD,60KD,51KD and 23KD were associated with antisperm antibody and immunological infertility.

  10. 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

    2016-01-01

    Chikungunya virus (CHIKV), a tropical pathogen, has re-emerged and has massive outbreaks abruptly all over the world. Containing many dominant epitopes, the envelope E2 protein of CHIKV has been explored for the vaccination or diagnosis. In the present study, the antigenicity of a recombinant expressed intrinsically disorder domain (IUD) of E2 was tested for the detection of the antibody against CHIKV through western blot method. The gene of the IUD of E2 was inserted into 2 different vectors and expressed as recombinant GST-E2 and recombinant MBP-E2 fusion protein, respectively. Two kinds of fusion proteins were tested with 30 CHIKV patient sera and 30 normal sera, respectively. Both proteins were detected by 25 patients sera (83.3%) and 1 normal serum (3.3%). This test showed a relatively high sensitivity and very high specificity of the recombinant E2 proteins to be used as diagnostic antigens against CHIKV infection. PMID:27180586

  11. 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.

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

    Science.gov (United States)

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

    2001-06-01

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

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

    Directory of Open Access Journals (Sweden)

    Samantha L Eaton

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

  14. 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.

  15. 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.

  16. 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.

  17. 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.

  18. 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.

  19. 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.

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

    Directory of Open Access Journals (Sweden)

    MORALES Olga Lucía

    2002-01-01

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

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

    Science.gov (United States)

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

    2014-06-01

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

  2. 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.

  3. 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.

  4. 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.

  5. 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

    2017-07-13

    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.

  6. 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.

  7. 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.

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

    Science.gov (United States)

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

    2015-01-01

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

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

    Science.gov (United States)

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

    1999-01-01

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

  10. 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.

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

    Science.gov (United States)

    Hashemi, Maryam; Madani, Rasool; Razmi, Nematollah

    2014-04-01

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

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

    Science.gov (United States)

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

    2007-07-01

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

  14. 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.

  15. 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.

  16. 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.

  17. Western blot confirmation of the H+/K+-ATPase proton pump in the human larynx and submandibular gland.

    Science.gov (United States)

    Altman, Kenneth W; Kinoshita, Yayoi; Tan, Melin; Burstein, David; Radosevich, James A

    2011-11-01

    The authors have previously demonstrated the H(+)/K(+)-ATPase (proton pump) in human larynx and lung glands via immunohistochemistry (IHC). The present hypothesis is that the proton pump is expressed in other seromucinous glands of the digestive tract that can be confirmed by IHC and Western blot analysis. Prospective controlled tissue analysis study. Academic medical institution. Ten anonymous fresh-frozen donor specimens were obtained, comprising 3 submandibular glands, 4 larynges, and 3 normal stomach specimens for control. Submandibular gland sections were immunostained with 2 monoclonal antibodies selectively reactive with α or β subunits of the H(+)/K(+)-ATPase. Western blot analysis was performed on all specimens. Consistent IHC staining was observed in the submandibular gland specimens for both α and β subunits. Western blot analysis revealed very strong expression for the stomach at 100 kDa, corresponding to the α protein, and weak but notable banding for all larynx and submandibular gland specimens. Similar findings were noted for the 60- to 80-kDa glycosylated β subunit protein, as well as the 52-kDa β subunit precursor for all specimens. The H(+)/K(+)-ATPase (proton) pump is present in the human larynx and submandibular gland although in much lower concentrations than in the stomach. Proton pump involvement in human aerodigestive seromucinous glands may have a role in protecting mucosa from acid environments (local or systemic), explain heightened laryngeal sensitivity in those patients with laryngopharyngeal reflux, and be a site of action for proton pump inhibitor pharmacotherapy.

  18. 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.

  19. 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.

  20. Affinity and Western blotting reveal homologies between ovine intervertebral disc serine proteinase inhibitory proteins and bovine pancreatic trypsin inhibitor.

    Science.gov (United States)

    Melrose, J; Shen, B; Ghosh, P

    2001-12-01

    The objective of this study was to assess any similarities between ovine intervertebral disc (IVD) serine proteinase inhibitory proteins (SPIs) and known mammalian IVD SPIs. Ovine IVDs were dissected into the annulus fibrosus and nucleus pulposus and the tissue finely diced then extracted with 4 M guanidine hydrochloride. The tissue extracts were subjected to caesium chloride density gradient ultracentrifugation to separate the large high buoyant density (rho > 1.5 g/mL) proteoglycans from the SPI proteins of low buoyant density (rho top two ultracentrifuge fractions containing the SPIs of interest were subjected to enzyme linked immunosorbent analysis (ELISA) and also examined by Western and Affinity blotting using an antibody to bovine pancreatic trypsin inhibitor and biotinylated trypsin respectively for detection and an alkaline phosphatase 5-bromo-4-chloro-3-indolyl phosphate/nitro blue tetrazolium system for visualisation. The major SPI proteins present in the Western and Affinity blots were 34-36 kDa species, minor 12 and 16, and 85 and 120 kDa species were also present. Qualitatively similar results were obtained for each respective tissue zone of the lumbar and lumbosacral disc specimens examined. Densitometric analysis of the major 34-36 kDa SPI bands visualised on Western and Affinity blots using NIH 1.61.1 image analysis software indicated that lumbar IVD samples contained higher levels of this SPI species than lumbosacral IVD samples. ELISA confirmed that lumbar IVD extracts contained quantitatively higher levels of BPTI equivalents per g of tissue extracted than lumbosacral IVDs. This study therefore has demonstrated that the ovine disc contains a range of SPI species which share some homology with bovine pancreatic trypsin inhibitor and in this respect are similar to SPIs previously demonstrated in canine IVDs.

  1. Transcription Factors Downstream of IL-4 and TGF-β Signals: Analysis by Quantitative PCR, Western Blot, and Flow Cytometry.

    Science.gov (United States)

    Sugimoto, Atsushi; Kawakami, Ryoji; Mikami, Norihisa

    2017-01-01

    IL-9-producing Th9 cell is a novel Th cell subset involved in type II allergic inflammations such as asthma. Th9 cells can be induced from naïve Th cells in the presence of IL-4 and TGF-β. It is also well established that downstream signals of IL-4 and TGF-β, including STAT6, IRF4, Smad, and PU.1, directly mediate IL-9 production in Th9 cells. In this chapter we describe the methods of flow cytometry, qPCR and western blot analysis to determine the expression or activation of these transcription factors downstream of IL-4 and TGF-β.

  2. 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).

  3. 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.

  4. [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.

  5. Analysis of Sperm Membrane Protein Relevant to Antisperm Antibody by Two-Dimensional Gel Electrophoresis and Western Blotting

    Institute of Scientific and Technical Information of China (English)

    Hao-fei WANG; Zhu-qiong XIANG; Yi-xing WANG

    2003-01-01

    Objective To identify the sperm membrane proteins that are associated with antisperm antibodyMethods Using antisperm antibody positive serum through unidimensional polyacrylamide gel electrophoresis and 2-dimensional gel electrophoresis followed by Western blot analysis to determine the molecular weights (MW) and isoelectric points (pI) of sperm membrane proteins that are associated with antisperm antibody.Results Eight kinds of MW with more than ten sperm membrane proteins can be recognized by antisperm antibody positive serum, of which the MWs and pI were 23 kD, 31 kD, 32 kD, 34 kD, 41 kD, 51 kD, 60 kD, 78 kD and 5.3, 5.5,5.7, 5.0, 5.3, 5.8, 6.0, 5.5~6.2, 4.6,5.1,5.5~5.8 respectively. The identification ratios of the sperm membrane proteins on 78 kD (60.7%), 60 kD (71.4%), 51 kD (14.9%) and 23 kD (14.29%) were higher.Conclusion The sperm membrane proteins with MW of 78 kD, 60 kD, 51 kD and 23 kD were associated with antisperm antibody and immunological infertility. Two-dimensional gel electrophoresis and Western blotting can precisely identify the sperm membrane proteins that are associated with antisperm antibody.

  6. Interaction of murine intestinal mast cell proteinase with inhibitors (serpins) in blood; analysis by SDS-PAGE and western blotting.

    Science.gov (United States)

    Irvine, J; Newlands, G F; Huntley, J F; Miller, H R

    1990-01-01

    The interaction of mouse intestinal mast cell proteinase (IMCP) with serine proteinase inhibitors (serpins) in blood was analysed: (i) by examining the capacity of the inhibitors in blood to block the binding of the irreversible serine esterase inhibitor [3H]diisopropyl fluorophosphate (DFP); (ii) by Western blotting. The binding of [3H]DFP to IMCP was blocked very rapidly by inhibitors in mouse serum and, by Western blotting, this inhibition was associated with the appearance of a 73,000 MW proteinase/inhibitor complex together with a series of higher (greater than 100,000) MW complexes. IMCP was not dissociated from these complexes when electrophoresed under reducing conditions, although prior heat treatment of mouse serum (60 for 30-160 min) abolished the formation of all proteinase/inhibitor complexes. Similarly, the activity of a 48,000 MW inhibitor of chymotrypsin was abolished by heat treatment. A titration experiment established that between 0.5 and 5 mg IMCP were inhibited per ml of serum. The properties and MW of the IMCP inhibitor complexes are typical of serpins and suggest that IMCP secreted during intestinal immunological reactions would be rapidly and irreversibly inactivated by plasma-derived inhibitors.

  7. 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.

  8. 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.

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

    Science.gov (United States)

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

    2015-12-01

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

  10. 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.

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

    Science.gov (United States)

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

    2014-10-01

    Large conductance Ca(2+)-activated K(+) (BK) channels consist of pore-forming α- and accessory β-subunits. There are four β-subunit subtypes (β1-β4), BK β1-subunit is specific for smooth muscle cells (SMC). Reduced BK β1-subunit expression is associated with SMC dysfunction in animal models of human disease, because downregulation of BK β1-subunit reduces channel activity and increases SMC contractility. Several anti-BK β1-subunit antibodies are commercially available; however, the specificity of most antibodies has not been tested or confirmed in the tissues from BK β1-subunit knockout (KO) mice. In this study, we tested the specificity and sensitivity of six commercially available antibodies from five manufacturers. We performed western blot analysis on BK β1-subunit enriched tissues (mesenteric arteries and colons) and non-SM tissue (cortex of kidney) from wild-type (WT) and BK β1-KO mice. We found that antibodies either detected protein bands of the appropriate molecular weight in tissues from both WT and BK β1-KO mice or failed to detect protein bands at the appropriate molecular weight in tissues from WT mice, suggesting that these antibodies may lack specificity for the BK β1-subunit. The absence of BK β1-subunit mRNA expression in arteries, colons, and kidneys from BK β1-KO mice was confirmed by RT-PCR analysis. We conclude that these commercially available antibodies might not be reliable tools for studying BK β1-subunit expression in murine tissues under the denaturing conditions that we have used. Data obtained using commercially available antibodies should be interpreted cautiously. Our studies underscore the importance of proper negative controls in western blot analyses. © 2014 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.

  12. 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.

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

    Directory of Open Access Journals (Sweden)

    Luiz Claudio Pereira Ribeiro

    2013-09-01

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

  14. 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.

  15. 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.

  16. 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.

  17. 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.

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

    Science.gov (United States)

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

    2012-04-30

    Horses serve as an intermediate host for several species of Sarcocystis, all of which utilize canids as the definitive host. Sarcocystis spp. infection and formation of latent sarcocysts in horses often appears to be subclinical, but morbidity can occur, especially when the parasite burden is large. A serological survey was conducted to determine the presence of antibodies against Sarcocystis spp. in seemingly healthy horses from the Galicia region of Spain. Western blot analyses using Sarcocystis neurona merozoites as heterologous antigen suggested greater than 80% seroprevalance of Sarcocystis spp. in a sample set of 138 horses. The serum samples were further tested with enzyme-linked immunosorbent assays (ELISAs) based on recombinant S. neurona-specific surface antigens (rSnSAGs). As expected for horses from the Eastern Hemisphere, less than 4% of the serum samples were positive when analyzed with either the rSnSAG2 or the rSnSAG4/3 ELISAs. An additional 246 horses were tested using the rSnSAG2 ELISA, which revealed that less than 3% of the 384 samples were seropositive. Collectively, the results of this serologic study suggested that a large proportion of horses from this region of Spain are exposed to Sarcocystis spp. Furthermore, the anti-Sarcocystis seroreactivity in these European horses could be clearly distinguished from anti-S. neurona antibodies using the rSnSAG2 and rSnSAG4/3 ELISAs. Copyright © 2011 Elsevier B.V. All rights reserved.

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

    Directory of Open Access Journals (Sweden)

    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.

  20. 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.

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

    Science.gov (United States)

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

    1995-01-01

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

  2. 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

  3. Evaluation of western blotting for the diagnosis of enzootic bovine leukemia Avaliação da técnica de western blot no diagnóstico da leucose enzoótica bovina

    Directory of Open Access Journals (Sweden)

    E.T. Gonzalez

    1999-08-01

    Full Text Available A western blotting (WB procedure has been developed for detecting antibodies to bovine leukosis virus (BLV in cattle sera. Two hundred and thirty three serum samples from naturally infected cattle with BLV virus and serial bleedings from experimentally BLV infected cows were used. An agar gel immunodiffusion test (AGID was used for comparing with the results obtained by WB. The AGID positive sera showed a different degree of reactivity by WB test against the two most important viral antigens (gp51 and p24, or against one of them. Other proteins (gp30, p15, p12 and p10 were not detected with any AGID positive sera, being observed occasionally three bands corresponding to the p24 protein. Using sera obtained by BLV experimental inoculation, the antibodies directed to p24 appeared early (between the 2nd and 4th week post inoculation and thereafter antibodies to gp51were detected in some animals. The analysis of field serum samples by AGID as compared to WB showed an agreement of 90.9%. Only 1.7% of sera were negative by AGID and positive by WB and 7.2% that were not conclusive by AGID and were defined by WB (4.2% as positive and 3.0% as negative.Um sistema de western blotting (WB foi desenvolvido para detecção de anticorpos contra o vírus da leucose em soros de bovinos. Foram utilizadas amostras de soros de 233 animais naturalmente infectados e soros de vacas experimentalmente infectadas. O teste de imunodifusão em ágar (AGID foi usado para comparação dos resultados. Graus diferentes de reatividade foram observados em soros positivos ao AGID, quando testados em WB frente a um ou aos dois antígenos mais importantes (gp51 e p24. Outras proteínas (gp30, p15, p12 e p10 não foram detectadas por nenhum soro positivo ao AGID, sendo que três bandas correspondentes à proteína p24 foram observadas ocasionalmente. Em soros obtidos por inoculação experimental, anticorpos contra a proteína p24 foram detectados entre a segunda e a quarta semanas

  4. Relative performance of Organon kit in comparison to Du Pont for confirmatory serological testing of HIV infection by western blot test in sera from blood donors.

    Science.gov (United States)

    Aggarwal, R K; Chatterjee, R; Chattopadhya, D; Kumari, S

    1992-06-01

    A total of 32 specimens with different categories of reactivity by Du Pont Western Blot kit comprising of specimens showing full spectrum of HIV-I antigen specific bands, 19 specimens showing total absence of bands and four specimens showing non-specific bands (without any interpretative importance) were subjected to Western Blot testing by Organon test. Of the nine specimens showing full spectrum of bands by Du Pont the correlation with Organon kit was 100 per cent based on WHO criteria. Four specimens with non-specific indeterminate band pattern by Du Pont failed to show any band in Organon kit, indicating that latter to be more specific.

  5. 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

  6. 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.

  7. 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.

  8. 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

    2017-03-11

    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.

  9. 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.

  10. 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.

  11. Estrogen receptors alpha and beta in rat placenta: detection by RT-PCR, real time PCR and Western blotting

    Directory of Open Access Journals (Sweden)

    Al-Bader Maie D

    2006-03-01

    Full Text Available Abstract Background High levels of estrogens during pregnancy not only retard placental and fetal growth but can lead to reproductive tract abnormalities in male progeny. Estrogens act through estrogen receptors (ER to modulate the transcription of target genes. These ER exist in two isoforms, ER alpha and ER beta and recently several variants of these isoforms have been identified. Methods The expressions of ER isoforms and variants have been studied in rat placenta at 16, 19 and 21 days gestation (dg. Gene expression was assessed using RT-PCR and real time PCR while protein expression was studied using Western blotting followed by immunodetection. Placental homogenates were probed with: a monoclonal antibody raised against the steroid binding domain of the ER alpha (ER alpha -S, a monoclonal antibody raised against the hinge region of ER alpha (ER alpha -H and a polyclonal antibody raised against the amino terminus of ER beta. Results ER alpha and ER beta mRNA and protein were detected from as early as 16 dg. Two PCR products were detected for ER alpha, one for the wild type ER alpha, and a smaller variant. Real time PCR results suggested the presence of a single product for ER beta. The antibodies used for detection of ER alpha protein both identified a single 67 kDa isoform; however a second 54 kDa band, which may be an ER alpha variant, was identified when using the ER alpha -H antibody. The abundance of both ER alpha bands decreased significantly between 16 and 19 dg. As for ER beta, four bands (76, 59, 54 and 41 kDa were detected. The abundance of the 59 and 54 kDa bands decreased significantly between 16 and 19 dg. Conclusion This study shows that both ER protein isoforms and their variants are present in rat placenta. The decrease in their expression near parturition suggests that the placenta may be relatively unresponsive to estrogens at this stage.

  12. 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...

  13. 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.

  14. 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

  15. 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

  16. 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.

  17. 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.

  18. 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.

  19. 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.

  20. Analysis of the Crude Antigen of Hymenolepis nana from Mice by SDS-PAGE and the Determination of Specific Antigens in Protein Structure by Western Blotting

    OpenAIRE

    GÖNENÇ, Bahadır

    2002-01-01

    Protein bands of crude antigens of Hymenolepis nana were determined by SDS-PAGE and Western blotting. Thirty Swiss albino mice were allotted into two groups of 15 each as positive (infected with H. nana) and negative (non-infected with H. nana) groups. The natural infections of H. nana and other helminths were determined by centrifugal flotation of faeces. After bleeding, the mice were necropsied and their guts were examined for H. nana and other intestinal helminths. Sera from mice were test...

  1. 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.

  2. 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

    2017-09-11

    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.

  3. Western blot analysis of a limited number of cells: a valuable adjunct to proteome analysis of paraffin wax-embedded, alcohol-fixed tissue after laser capture microdissection.

    Science.gov (United States)

    Martinet, Wim; Abbeloos, Vanessa; Van Acker, Nathalie; De Meyer, Guido R Y; Herman, Arnold G; Kockx, Mark M

    2004-03-01

    In recent years, laser capture microdissection (LCM) has been used successfully to obtain distinct populations of cells for subsequent molecular analysis. Because of the limited sample availability and the absence of in vitro amplification steps for proteins, the use of LCM for proteome analysis largely depends on highly sensitive protein detection methods. In this study, a western blot protocol was developed and validated for the detection of beta-actin and the moderately expressed cell death protein caspase-3 in small numbers of cells. Initially, cultured human U937 monocytes and whole sections of paraffin wax-embedded, alcohol-fixed human tonsils were used to optimize protein electrophoresis and western blotting conditions. High-performance NuPAGE Bis-Tris gels in combination with high-quality transfer membranes, optimized antibody concentrations, and a sensitive chemiluminescent substrate provided a strong signal for beta-actin with approximately 500 U937 cells. In the same way, procaspase-3 could be identified with approximately 1000 cells. Similar results were obtained with germinal centre cells that were procured from paraffin wax-embedded, alcohol-fixed human tonsils by LCM. Treatment of U937 cells with etoposide rapidly induced cell death and allowed the detection of active caspase-3 with approximately 2500 cells (0.8 pg of protein). The findings of this study suggest that western blotting is a valuable adjunct to proteome analysis of LCM procured cells.

  4. 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

  5. Cytomegalovirus Destruction of Focal Adhesions Revealed in a High-Throughput Western Blot Analysis of Cellular Protein Expression† ▿

    OpenAIRE

    Stanton, Richard James; McSharry, Brian Patrick; Rickards, Carole Ruth; Wang, Edward Chung Yern; Tomasec, Peter; Wilkinson, Gavin William Grahame

    2007-01-01

    Human cytomegalovirus (HCMV) systematically manages the expression of cellular functions, rather than exerting the global shutoff of host cell protein synthesis commonly observed with other herpesviruses during the lytic cycle. While microarray technology has provided remarkable insights into viral control of the cellular transcriptome, HCMV is known to encode multiple mechanisms for posttranscriptional and posttranslation regulation of cellular gene expression. High-throughput Western blotti...

  6. 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.’

  7. Study of a viral-dual infection in rainbow trout (Oncorhynchus mykiss) by seroneutralization, western blot and polymerase chain reaction assays.

    Science.gov (United States)

    Rodríguez, S; Vilas, M P; Alonso, M; Pérez, S I

    1995-12-01

    Viral-dual infections in fish are of interest to aquaculture practices but they are rarely described and studied. Several methods were applied in this work to demonstrate a case of coinfection in a reared rainbow trout (Oncorhynchus mykiss) population. Inoculation in cell cultures and cross-neutralization tests were the standard procedures that made it possible to isolate and identify a birnavirus, the infectious pancreatic necrosis virus (IPNV), and suspect of a second virus. Western blotting with both polyclonal and monoclonal antibodies, and reverse transcriptional-polymerase chain reaction (RT-PCR) demonstrate coexistence of both, IPNV and a rhabdovirus.

  8. 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.

  9. APPLICATION OF WESTERN BLOTTING TECHNIQUE FOR EVALUATING THE EXPRESSION OF VASOPRESSIN RECEPTORS IN THE HEART CELLS; IMPORTANCE IN THE CARDIOVASCULAR SYSTEM

    Directory of Open Access Journals (Sweden)

    Manoj G Tyagi

    2012-08-01

    Full Text Available Vasopressin, a posterior pituitary hormone is responsible for water reabsorption by the kidneys and maintenance of cardio-vascular homeostasis. Vasopressin receptors are characterized as VR 1 (V1a, VR2 (V2, and VR3 (V1b. VR1, which is abundant in vascular smooth muscles, causes vasoconstriction by increasing intracellular calcium via the phosphatidylinositol bisphosphonate pathway and a positive inotropic effect in cardiac muscle. VR2 has also been shown to be expressed in the heart. There is emerging role for vasopressin receptors in health and disease. This study describes the application of Western blotting to elucidate the importance of vasopressin receptors in the heart cells.

  10. 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.

  11. 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.

  12. 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.

  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. Philosophy at Cambridge

    OpenAIRE

    2011-01-01

    Newsletter of the Philosophy Faculty. Articles by: Simon Blackburn, 'From the Chair' ; Nick Treanor, 'Inaugural lecture: What is distinctive about human thought?' ; Clare Chambers, 'Political Philosophy at Cambridge' ; Alexis Papazoglou,'Aspects of philosophy at Cambridge' ; Peter Smith, 'Principia at 100' ; Nigel Crisp, 'Turning the World Upside Down' ; Cain Todd, 'Fiction, Emotion, Imagination'; Fraser MacBride, 'Philosophy, St John's, Cambridge, 1986.'

  15. 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.

  16. 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.

  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

    physiology where the intervention has been suggested to impede normalization (ageing, muscle atrophy, and different muscle fiber type composition). The linearity of signal and the methodological variation coefficient was obtained. Furthermore, the inter- and intraindividual variation in signals obtained from...... SF and RP was measured in relation to ageing, muscle atrophy, and different muscle fiber type composition, respectively. A stronger linearity of SF and β-actin compared with GAPDH and α-tubulin was observed. The methodological variation was relatively low in all four methods (4-11%). Protein level...... 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. 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.

  19. 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.

  20. Pacientes neurológicos del noroeste del Perú con serología positiva por Western Blot a la larva de Taenia solium

    Directory of Open Access Journals (Sweden)

    Hermes Escalante A

    2004-04-01

    Full Text Available Objetivos: Determinar la frecuencia de pacientes con sintomatología neurológica de la zona noroeste del Perú que presentan serología positiva por Western Blot a la larva de Taenia solium. Material y Métodos: El estudio se realizó en 3515 pacientes de cero a noventa años de edad y de ambos sexos con sintomatología neurológica, procedentes de los departamentos de Piura, Lambayeque, Cajamarca, Ancash, La Libertad y algunas poblaciones de la zona selvática (5 a 10° LS; 77 a 82° LO. Dichos pacientes fueron atendidos en consultorios privados y en los servicios de Neurología de los Hospitales Belén y Regional de Trujillo desde enero de 1997 hasta diciembre de 2000. De cada uno de los pacientes se obtuvo una muestra sanguínea, cuyo suero fue procesado por la técnica de Western Blot, la cual tiene una sensibilidad de 91% y una especificidad de 100%. Resultados: Se detectó serología positiva en 562 pacientes, la cual representa una prevalencia de 16%, siendo el sexo masculino el que presentó mayor porcentaje (58,4%. Los grupos etáreos con mayor frecuencia de serología positiva fueron los de 41 a 50 años (18,7% y 31 a 40 años (17,4%. Los mayores porcentajes de serología positiva se obtuvo en pacientes procedentes de Piura y Lambayeque. Conclusiones: Los pacientes estudiados presentan una alta frecuencia de serología positiva a la larva de T. solium.

  1. 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

  2. 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.

  3. Effects of tetracyclines on neutrophil, gingival, and salivary collagenases. A functional and western-blot assessment with special reference to their cellular sources in periodontal diseases.

    Science.gov (United States)

    Sorsa, T; Ding, Y; Salo, T; Lauhio, A; Teronen, O; Ingman, T; Ohtani, H; Andoh, N; Takeha, S; Konttinen, Y T

    1994-09-06

    The characterization and regulation of matrix metalloproteinases (MMPs) have been studied to determine their role(s) in periodontal tissue destruction. Progress in elucidating the roles of MMPs in periodontal tissue destruction has led to a new concept involving the chemotherapeutic inhibition on MMPs, a therapeutic strategy which less than a decade ago was considered "a difficult and perhaps impossible task." Tetracyclines/doxycycline (DOXY) and their chemically modified nonantimicrobial derivatives (CMTs) are known to inhibit the matrix metalloproteinases, especially preferring human neutrophil collagenase (MMP-8), and prevent the oxidative activation of procollagenases. We characterized by Western blotting the molecular forms and cellular sources of gingival tissue, dental plaque, gingival crevicular fluid (GCF), and salivary MMPs associated with periodontitis. Also the molecular forms of tissue inhibitors of matrix metalloproteinases (TIMP-1 and TIMP-2) in periodontitis were studied by Western blot. Neutrophil (PMN)-derived MMPs were found to predominate in periodontitis, and phospholipase C present in increased amounts in periodontitis sites was found to be a potential inducer of PMN degranulation. We further studied the effects of DOXY on molecular forms of different latent and active MMPs purified from different cellular sources (PMNs, fibroblasts, keratinocytes) and present in vivo in oral exudates (gingival extracts, GCF, and saliva). DOXY inhibition of activated (oxidatively or proteolytically) MMPs were not associated with MMP fragmentation. Michaelis-Menten plots of initial rates of degradation of soluble type I collagen revealed an apparent Km value of 0.3-0.6 microM for MMP-8, and 75 microM DOXY inhibited MMP-8 in a manner which did not result in changes in apparent Km value but did prevent the initial degradation reaching Vmax providing evidence for noncompetitive inhibition. Treatment of patients with long-term DOXY medication results in decreased

  4. 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.

  5. 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.

  6. 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

    2017-08-18

    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.

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

    Science.gov (United States)

    Hood-Degrenier, Jennifer K

    2008-01-01

    The movement of newly synthesized proteins through the endomembrane system of eukaryotic cells, often referred to generally as the secretory pathway, is a topic covered in most intermediate-level undergraduate cell biology courses. An article previously published in this journal described a laboratory exercise in which yeast mutants defective in two distinct steps of protein secretion were differentiated using a genetic reporter designed specifically to identify defects in the first step of the pathway, the insertion of proteins into the endoplasmic reticulum (Vallen, 2002). We have developed two versions of a Western blotting assay that serves as a second way of distinguishing the two secretory mutants, which we pair with the genetic assay in a 3-wk laboratory module. A quiz administered before and after students participated in the lab activities revealed significant postlab gains in their understanding of the secretory pathway and experimental techniques used to study it. A second survey administered at the end of the lab module assessed student perceptions of the efficacy of the lab activities; the results of this survey indicated that the experiments were successful in meeting a set of educational goals defined by the instructor.

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

    Science.gov (United States)

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

    2014-05-01

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

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

    Directory of Open Access Journals (Sweden)

    Terezinha de Jesus Teixeira Santos

    2003-06-01

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

  10. 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”.

  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. The University of Cambridge

    Institute of Scientific and Technical Information of China (English)

    郭大任

    2007-01-01

    The University of Cambridge was founded in 1209,just a few decades after Oxford University,making it the second oldest university in the English-speaking world.Like“the other place”, the university is made up of 31 colleges and more than 100 departments,catering for around 15,500 students.Cambridge has a worldwide reputation for outstanding

  13. Cambridge Scientific Abstracts

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    正Meteorological and Environmental Research has been included by Cambridge Scientific Abstracts (CSA) since 2011. CSA is a retrieval system published by Cambridge Information Group. CSA was founded in the late 1950's,and became part of the CIG family in 1971. CSA's original mission was publishing secondary source materials relating to the physical sciences. Completely

  14. Cambridge Scientific Abstracts

    Institute of Scientific and Technical Information of China (English)

    2012-01-01

    Meteorological and Environmental Research has been included by Cambridge Scientific Abstracts (CSA) since 2011. CSA is a retrieval system published by Cambridge Information Group. CSA was founded in the late 1950’s,and became part of the CIG family in 1971. CSA’s original mission was publishing secondary source materials relating to the physical sciences. Completely

  15. 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

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

    Directory of Open Access Journals (Sweden)

    Stephenie D Prokopec

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

  17. 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.

  18. 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.

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

    Directory of Open Access Journals (Sweden)

    Tetsuya Okuda

    2017-02-01

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

  20. 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.

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

    Directory of Open Access Journals (Sweden)

    Camila Cánepa

    2015-10-01

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

  2. 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

  3. 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...

  4. 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.

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

    Science.gov (United States)

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

    2013-10-01

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

  6. 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.

  7. 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

  8. 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

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

    Directory of Open Access Journals (Sweden)

    Neuza Satomi SATO

    1999-03-01

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

  10. Cambridge IGCSE computer science

    CERN Document Server

    Watson, Dave; Konrad, Nina

    2015-01-01

    Endorsed by Cambridge International Examinations. Develop your students computational thinking and programming skills with complete coverage of the latest syllabus (0478) from experienced examiners and teachers. - Includes a Student CD-ROM with interactive tests, based on the short answer questions from both papers - Follows the order of the syllabus exactly, ensuring complete coverage - Introduces students to self-learning exercises, helping them learn how to use their knowledge in new scenarios This syllabus is for first examination from 2015.

  11. 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 membran...

  12. Problem-Solving Test: Southwestern Blotting

    Science.gov (United States)

    Szeberényi, József

    2014-01-01

    Terms to be familiar with before you start to solve the test: Southern blotting, Western blotting, restriction endonucleases, agarose gel electrophoresis, nitrocellulose filter, molecular hybridization, polyacrylamide gel electrophoresis, proto-oncogene, c-abl, Src-homology domains, tyrosine protein kinase, nuclear localization signal, cDNA,…

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

    Science.gov (United States)

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

    2007-12-01

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

  14. Cambridge technicals level 3 IT

    CERN Document Server

    Ellis, Victoria; Middleton, Saundra

    2016-01-01

    Support your teaching of the new Cambridge Technicals 2016 suite with Cambridge Technical Level 3 IT, developed in partnership between OCR and Hodder Education; this textbook covers each specialist pathway and ensures your ability to deliver a flexible course that is both vocationally focused and academically thorough. Cambridge Technical Level 3 IT is matched exactly to the new specification and follows specialist pathways in IT Infrastructure Technician, Emerging Digital Technology Practitioner, Application Developer, and Data Analyst. - Ensures effective teaching of each specialis

  15. The Cambridge Experiment

    Directory of Open Access Journals (Sweden)

    Marco Iuliano

    2014-09-01

    Full Text Available Since the latter part of 19th century photography has played a central role in the development of architecture for its persuasive visual impact. But, despite this clear interaction, there is still reluctance from scholars in accepting less rigid approaches to the two disciplines. Indeed, the combination of the subjects, with the necessary rigour, can open up new and effective horizons for architectural history, with a potential influence on the perceived reality: this could gradually establish attention towards less known heritage. In the case we present here, by means of a provocative exhibition on Cambridge’s buildings after the Second World War, we have used photography to re-evaluate modern architecture. Cambridge in Concrete. Images from the RIBA British Architectural Library Photographs Collection, was held on the occasion of the University of Cambridge Department of Architecture’s Centenary (1912-2012. The cues for our task were contained in the collections of the Royal Institute of British Architects: the photographic archive is the world’s biggest holding of architectural images which, since 2012, has been renamed in honour of Robert Elwall (1953-2012, first curator of the collection. As part of the exhibition we published a limited edition catalogue; we have here revisited, combined and enlarged our original essays.

  16. Comparison analysis between Western blot method and Magnetic particle Chemiluminescence method detection SLE autoantibody%免疫印迹法和磁微粒化学发光法检测SLE自身抗体的比对分析

    Institute of Scientific and Technical Information of China (English)

    周厚清

    2016-01-01

    目的:比较免疫印迹法(Western blot)和磁微粒化学发光法(Magnetic particle Chemiluminescence method)检测系统性红斑狼疮(SLE)自身抗体的一致性。方法:用免疫印迹法和磁微粒化学发光法对123份血清进行检测,其中系统性红斑狼疮阳性患者63例,健康体检者60例,用卡方检验对两种方法结果进行比较分析。结果:免疫印迹法和磁微粒化学发光法中抗Sm抗体阳性率分别为27.0%和14.3%、抗RNP/Sm阳性率分别为54.0%和42.9%、dsDNA阳性率分别为41.3%和30.2%、抗核小体抗体阳性率分别为31.7%和20.6%、抗核糖体P蛋白抗体阳性率分别为28.6%和11.1%。两种方法的准确性无显著性差异(>0.05)总符合率为80.6%,Kappa值为0.669。结论:免疫印迹法和磁微粒化学发光法检测系统性红斑狼疮自身抗体的检测结果具有可靠性,较好的符合率和准确性,磁微粒化学发光法可以作为简便、快速、经济的检测方法应用于系统性红斑狼疮自身抗体的检测。%Objective: The present study aims to compare Western blot and Magnetic particle Chemiluminescence method consistency on autoimmune antibody detection of systemic lupus erythematosus. Methods: Western blot assay and Magnetic particle Chemiluminescence method were used to detect autoimmune antibody from 123 serum samples, of which 63 patients with systemic lupus erythematosus, the others from 60 healthy persons. The results of the two methods were compared by X2 test. Results: Anti-Sm antibody positive rate of Western blot and Magnetic particle Chemiluminescence method were 27.0 and 14.3 percent respectively. Anti- RNP/Sm antibody positive rates were 39.7% and 42.9% respectively. Anti- ds- DNA antibody positive rates were 41.3% and 30.2% respectively. Anti- nucleosome antibody positive rates were 31.7% and 20.6% respectively. Anti- ribosomal protein antibody positive rates were 28.6% and 11.1% respectively. The accuracy

  17. International Commercial Contracts, by Giuditta Cordero Moss. (Cambridge: Cambridge University Press, 2014)

    DEFF Research Database (Denmark)

    Lando, Ole

    2015-01-01

    Review of: Giuditta Cordero Moss, International Commercial Contracts. Cambridge: Cambridge University Press, 2014. XV + 329 pages. ISBN: 9781107684713......Review of: Giuditta Cordero Moss, International Commercial Contracts. Cambridge: Cambridge University Press, 2014. XV + 329 pages. ISBN: 9781107684713...

  18. Cambridge IGCSE computer science workbook

    CERN Document Server

    Watson, David; Guinness, Paul; Nagle, Garrett

    2016-01-01

    Student workbook to support students throughout their Cambridge IGCSE Computer Science course, with plenty of extra practice questions and activities. Accompanies the Cambridge IGCSE Computer Science student text book, following the contents and chapter order. Students write their answers in the book and keep it for study in class or at home throughout their course.

  19. 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...

  20. 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.

  1. 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.

  2. The Cambridge Structural Database.

    Science.gov (United States)

    Groom, Colin R; Bruno, Ian J; Lightfoot, Matthew P; Ward, Suzanna C

    2016-04-01

    The Cambridge Structural Database (CSD) contains a complete record of all published organic and metal-organic small-molecule crystal structures. The database has been in operation for over 50 years and continues to be the primary means of sharing structural chemistry data and knowledge across disciplines. As well as structures that are made public to support scientific articles, it includes many structures published directly as CSD Communications. All structures are processed both computationally and by expert structural chemistry editors prior to entering the database. A key component of this processing is the reliable association of the chemical identity of the structure studied with the experimental data. This important step helps ensure that data is widely discoverable and readily reusable. Content is further enriched through selective inclusion of additional experimental data. Entries are available to anyone through free CSD community web services. Linking services developed and maintained by the CCDC, combined with the use of standard identifiers, facilitate discovery from other resources. Data can also be accessed through CCDC and third party software applications and through an application programming interface.

  3. Clark and Prehistory at Cambridge

    Directory of Open Access Journals (Sweden)

    Pamela Jane Smith

    1996-05-01

    Full Text Available If honours and titles give measure of a man, then Professor Sir Grahame Clark was indeed important. Faculty Assistant Lecturer in the Faculty of Archaeology and Anthropology at Cambridge University from 1935-46, University Lecturer 1946-52, Disney Professor of Archaeology 1952-74, Head of the Department of Archaeol­ogy and Anthropology 1956-61 and 1968-71, Fellow of Peterhouse, Cambridge 1950-73, Master of Peterhouse 1973-80, he was a visiting lecturer at diverse universities; appointed CBE in 1971, he received many awards includ­ing the prestigious Erasmus Prize for 1990, presented by Prince Bernhard of the Netherlands, for his "long and inspiring devotion to prehistory" (Scarre 1991:10; and in June 1992, he was knighted. Yet well before fame and position were rewards, Clark made major contributions to the establishment of prehis­tory as an academic subject at Cambridge University. Cambridge was the first and, for many years, only British university granting an undergraduate degree which offered prehistory as a specialization. "The development of postgraduate research in prehistoric archaeology at Cambridge had to wait on the provision of undergraduate teaching;' Clark (1989b: 6 recently observed. The "faculty was the only one in Britain producing a flow of graduates in prehistoric archaeology" (Clark 1989a: 53.

  4. Cambridge checkpoint English workbook 1

    CERN Document Server

    Reynolds, John

    2013-01-01

    This Workbook supports our bestselling Checkpoint English series, with exercises specifically matched to the Cambridge Progression tests and the Checkpoint English tests. - Offers plenty of additional questions for use in class or as homework. - Includes clearly identified questions on grammar and punctuation, comprehension, use of language and essay planning. - Follows the structure of the relevant textbook to ensure a thorough understanding of all aspects of the course. - Provides a space for Students to write their answers. This Workbook is matched to the Cambridge Secondary 1 Curriculum Fr

  5. Cambridge IGCSE English first language

    CERN Document Server

    Reynolds, John

    2013-01-01

    Revised edition for the 2015 syllabus to help your students prepare for their examination and enhance their enjoyment of English. This title has been written for the revised Cambridge IGCSE First Language English (0500 and 0522) syllabuses, for first teaching from 2013. ? Develops the skills necessary to become a better reader and writer. ? Offers detailed advice and preparation for the examination. ? Teaches skills for successful writing of essays and coursework assignment. We are working with Cambridge International Examinations to gain endorsement for this title.

  6. 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

  7. 亚洲带绦虫囊尾蚴特异性抗原筛选和鉴定%Screen and detection of specific antigens of Taenia asiatica cysticercus with two dimensional electrophoresis, western blotting and MS/MS spectrometry

    Institute of Scientific and Technical Information of China (English)

    方文; 肖靓靓; 包怀恩; 牟荣

    2011-01-01

    Objective To screen and identify specific antigens of Taenia asiatica cysticercus and to predict the function of target proteins using bioinformatics method. Methods Six three-way cross hybrid pigs were randomly divided into experimental group and control group and each experimental pig was infected with 84 000 eggs. Serum samples were collected 40 days after the infection. The total protein of Taenia asiatica cysticercus was separated by two dimensional electrophoresis and western blotting was performed to find out distinct antigens. Proteins from the two groups were identified by electrospray ion trap mass spectrometry(ESI-Trap MS). Results Totally 204 ± 11 spots were detected through Coomassie brilliant blue stained gels with relative molecular weight( Mr) of 14 400 -94 000 kDa and isoelectric point(pl) of 3. 0 - 10. 0. Western-blotting showed 6 specific antigen spots with pool sera of infected pigs. Two of the 6 antigens with known functions were ascribed to ATPase family gene 3-like protein 2 and ATP synthase beta subunit. Conclusion Two specific antigens of Taenia asiatica cysticercus have been identified.%目的 对亚洲带绦虫囊尾蚴的特异性抗原进行筛选、鉴定和生物信息学分析.方法 6头20d龄三元杂交乳猪随机分为实验组和对照组,实验组每头灌胃亚洲带绦虫虫卵悬液8.4万个;20 d后,分别制备实验组及对照组心脏血混合血清;收集、制备实验组亚洲带绦虫囊尾蚴蛋白并进行双向电泳分析,将凝胶蛋白斑点转移至聚偏氟乙烯膜,用实验组及对照组混合血清(均为1∶10)为一抗,羊抗猪辣根过氧化物酶- IgG(1∶200)为二抗,进行蛋白质印迹分析和质谱鉴定.结果 双向电泳凝胶共检测到( 204±11)个蛋白质斑点,相对分子质量为14 400 ~94 000 kPa,等电点(pI)为3.0~10.0;筛选出6个特异性抗原,经质谱鉴定出其中2个蛋白,分别为ATP酶管家基因3-类蛋白2(AT-Pase family gene 3-like protein 2)

  8. 33 CFR 117.549 - Cambridge Harbor.

    Science.gov (United States)

    2010-07-01

    ... 33 Navigation and Navigable Waters 1 2010-07-01 2010-07-01 false Cambridge Harbor. 117.549 Section... DRAWBRIDGE OPERATION REGULATIONS Specific Requirements Maryland § 117.549 Cambridge Harbor. The draw of the S342 bridge, mile 0.1 at Cambridge, shall open on signal from 6 a.m. to 8 p.m.; except that, from...

  9. Cambridge checkpoint English workbook 2

    CERN Document Server

    Reynolds, John

    2014-01-01

    Build confidence and understanding throughout the year with hundreds of additional practice questions. This Workbook supports our bestselling Checkpoint series, with exercises specifically matched to the Cambridge Progression tests and the Checkpoint tests. - Develops understanding and builds confidence ahead of assessment with exercises matched to the tests - Ensures a thorough understanding of all aspects of the course by following the structure of the relevant textbook - Saves planning time with exercises that are suitable for use in class or as homework This Workbook is

  10. Cambridge checkpoint English workbook 3

    CERN Document Server

    Reynolds, John

    2014-01-01

    Build confidence and understanding throughout the year with hundreds of additional practice questions. This Workbook supports our bestselling Checkpoint series, with exercises specifically matched to the Cambridge Progression tests and the Checkpoint tests. - Develops understanding and builds confidence ahead of assessment with exercises matched to the tests - Ensures a thorough understanding of all aspects of the course by following the structure of the relevant textbook - Saves planning time with exercises that are suitable for use in class or as homework This Workbook is

  11. Honorary Degree Congregation in Cambridge

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    One day in mid-June 2006 when I was on a visit to the Countryside Restoration Trust at Barton near Cambridge, Mr. Christopher Stevenson, the director of Program of Events for Newcomers & Academic Visitors, gave me a letter enclosing a notice and a ticket I booked nearly two months earlier. He told me that I was very lucky because a strictly limited number of tickets had been allocated to academic visitors. It was a ticket to admit me to the Honorary Degree Congregation and to the reception afterwards on Tuesday 27 June.

  12. Michael Byers, International Law and the Arctic (Cambridge: Cambridge Studies in International and Comparative law, Cambridge University Press, 2013

    Directory of Open Access Journals (Sweden)

    Rachael L. Johnstone

    2014-03-01

    Full Text Available A review of: Michael Byers, International Law and the Arctic, Cambridge Studies in International and Comparative law, Cambridge University Press, 2013. pp. 314 + xviii, 65.00 GBP (hardcover; 16.56GBP (kindle edition ISBN: 9781107042759 ISBN: 9781107042759

  13. The Cambridge double star atlas

    CERN Document Server

    MacEvoy, Bruce; Mullaney, James; Tirion, Wil; Mullaney, James

    2015-01-01

    The Cambridge Double Star Atlas is back! It is the first and only atlas of physical double stars that can be viewed with amateur astronomical instruments. Completely rewritten, this new edition explains the latest research into double stars, and looks at the equipment, techniques and opportunities that will enable you to discover, observe and measure them. The target list has been completely revised and extended to 2500 binary or multiple systems. Each system is described with the most recent and accurate data from the authoritative Washington Double Star Catalog, including the HD and SAO numbers that are most useful in our digital age. Hundreds of remarks explain the attributes of local, rapidly changing, often measured or known orbital systems. The color atlas charts by Wil Tirion have been updated to help you easily find and identify the target systems, as well as other deep-sky objects. This is an essential reference for double star observers..

  14. Trial access to Cambridge University Press ebooks

    CERN Multimedia

    CERN Library

    2011-01-01

    From 1 August till 31 October, CERN users are invited to enjoy a trial access to all Cambridge University Press electronic books: http://ebooks.cambridge.org/. Please don't hesitate to send feedback to library.desk@cern.ch.

  15. 新孢子虫和弓形虫ELISA及western blot检测方法的建立及应用%Establishment and application of indirect ELISA assays and western blot to detect antibodies agaist Neospora caninum and Toxoplasma gondii

    Institute of Scientific and Technical Information of China (English)

    陈亮; 闫双; 刘启生; 曹雯丽; 王真; 巴音查汗

    2012-01-01

    为建立牛新孢子虫和弓形虫的免疫学检测方法,并调查新疆部分地区牛新孢子虫和弓形虫的感染情况,本研究应用纯化的新孢子虫重组蛋白SRS2 (NcSRS2)和弓形虫重组蛋白SAG2 (TgSAG2)作为包被抗原,分别建立新孢子虫和弓形虫的ELISA和western blot血清学检测方法,并进行特异性和重复性试验,以其检测662份疑似样品,并与商品化试剂盒检测结果比较验证.特异性和重复性试验结果表明,建立的方法特异性强、重复性良好.采用建立的两种ELISA方法对662份临床样品的检测结果表明,新孢子虫和弓形虫的抗体阳性率分别为13.44%(89/662)和5.29%(35/662);与IDEXX试剂盒和永辉试剂盒的符合率分别为94.11%和95.92%.此外,western blot检测的新孢子虫和弓形虫抗体阳性率分别为5.14%(34/662)和3.17%(21/662);与建立的ELISA检测方法的符合率分别为91.69%和97.89%.本研究为分析奶牛流产的原因提供了一定的依据.%Neospora caninum and Toxoplasma gondii are closely related protozoan parasites which causes of abortion and congenital disease in ruminants. To establish the methods for detection of the antibodies against N. caninum and T. gondii in bovine, The indirect ELISAs and western blot assay were developed based on purified recombinant protein SRS2 of N. cammim and SAG2 of T. gondii, respectively. The results show that these methods were highly specificity and repeatability. A total of 662 bovine serum samples were tested and the positive rates for N. caninum and T. gondii were 13.44% (89/662) and 5.29% (35/662) detected by the developed ELISAs, which were 94.11% and 95.92% agreements with TDEXX' N. caninum Antibody Test Kit and Yong hui' T. gondii Antibody Test Kit, respectively. In addition, the positive rates for N. caninum and T. gondii were 5.14% (34/662) and 3.17% (21/662) using western blot, and the agreements with the developed ELISAs were 91.69% and 97

  16. Cambridge community Optometry Glaucoma Scheme.

    Science.gov (United States)

    Keenan, Jonathan; Shahid, Humma; Bourne, Rupert R; White, Andrew J; Martin, Keith R

    2015-04-01

    With a higher life expectancy, there is an increased demand for hospital glaucoma services in the United Kingdom. The Cambridge community Optometry Glaucoma Scheme (COGS) was initiated in 2010, where new referrals for suspected glaucoma are evaluated by community optometrists with a special interest in glaucoma, with virtual electronic review and validation by a consultant ophthalmologist with special interest in glaucoma. 1733 patients were evaluated by this scheme between 2010 and 2013. Clinical assessment is performed by the optometrist at a remote site. Goldmann applanation tonometry, pachymetry, monoscopic colour optic disc photographs and automated Humphrey visual field testing are performed. A clinical decision is made as to whether a patient has glaucoma or is a suspect, and referred on or discharged as a false positive referral. The clinical findings, optic disc photographs and visual field test results are transmitted electronically for virtual review by a consultant ophthalmologist. The number of false positive referrals from initial referral into the scheme. Of the patients, 46.6% were discharged at assessment and a further 5.7% were discharged following virtual review. Of the patients initially discharged, 2.8% were recalled following virtual review. Following assessment at the hospital, a further 10.5% were discharged after a single visit. The COGS community-based glaucoma screening programme is a safe and effective way of evaluating glaucoma referrals in the community and reducing false-positive referrals for glaucoma into the hospital system. © 2014 Royal Australian and New Zealand College of Ophthalmologists.

  17. Cambridge checkpoint English revision guide for the Cambridge secondary 1 test

    CERN Document Server

    Reynolds, John

    2013-01-01

    With Checkpoint English Revision Guide for the Cambridge Secondary 1 test you can aim for the best grade with the help of relevant and accessible notes, examiner advice plus questions and answers on each key topic. - Clear explanations of every topic covered in the Cambridge Secondary 1 Checkpoint English syllabus. - Builds revision skills you need for success in the test. - Exam tips wirtten by test setters and examiners giving you their expert advice. This text has not been through the Cambridge endorsement process.

  18. Cambridge checkpoint maths revision guide for the Cambridge secondary 1 test

    CERN Document Server

    Smith, Alan

    2013-01-01

    With Checkpoint Maths Revision Guide for the Cambridge Secondary 1 test you can aim for the best grade with the help of relevant and accessible notes, examiner advice plus questions and answers on each key topic. - Clear explanations of every topic covered in the Cambridge Secondary 1 Checkpoint Maths syllabus. - Builds revision skills you need for success in the test. - Exam tips wirtten by test setters and examiners giving you their expert advice. This text has not been through the Cambridge endorsement process.

  19. 孕产妇HIV抗体筛查阳性与免疫印迹试验对比%Comparison of Positive of HIV Antibody Screening and Western Blot Test in Pregnant Women

    Institute of Scientific and Technical Information of China (English)

    朱厚宏; 王晨笛; 刘杨; 张祁; 孟建彤; 黄薇

    2011-01-01

    Objective To explore the relationship between the results of HIV antibody screening test and Westem Blot test ( WB) in pregnant women. Methods Thirty three positive samples from pregnant women were tested with the ELISA and Westem blot test (WB). Those with indefinite result by WB test were re-tested 3 and 6 months later, and HIV-1 viral load testing ( VL) was carried out at 6th month. The results of ELISA and the WB test were analyzed. Result The 33 samples were all positive by ELISA and 20 were HIV-1 positive by WB test. The ELISA and the WB test positive coincidence rate was 60. 61 % (20/33). Of 20 samples 0were with S/CO value more than 8, the WB test results of HIV-1 antibody were all positive. Of 13 samples were with S/CO value less than 1 , 5 were negative and 8 were indefinite in WB test. The HIV-1 viral load test result at 6th month was less than the minimum detection limit (50 IU/ml ) . Conclusion The false positive in HIV antibody screening test (ELISA) was observed. The false positive sample were in the WB test negative and indefinite ones. ELISA and the WB test positive coincidence rate increased with the S/CO value of ELISA. The HIV WB test indefinite result in pregnant women was mostly due to non-specific immune response.%目的 探讨孕产妇人类免疫缺陷病毒(HIV)抗体筛查试验阳性与免疫印迹试验(WB)结果的关系.方法 对33份孕产妇HIV抗体待复查样本,进行酶联免疫吸附试验(ELISA )及免疫印迹试验(WB),对WB试验结果不确定者进行3个月、6个月随访检测,6个月随访样本补充HIV-1病毒载量检测,分析ELISA检测与WB试验检测结果.结果 33份样本ELISA复检均呈阳性,免疫印迹试验(WB)中20份确认为HIV-1抗体阳性,ELISA检测与WB试验阳性符合率为60.61%(20/33); S/CO值>8的20份样本,WB试验结果均为HIV-1抗体阳性,I < S/CO值<8的13份样本,WB试验5份为HIV抗体阴性,8份为HIV抗体不确定;6个月随访样本HIV-1病毒载量检测,

  20. 77 FR 3118 - Security Zone; Choptank River and Cambridge Channel, Cambridge, MD

    Science.gov (United States)

    2012-01-23

    ... 3118-3121] [FR Doc No: 2012-1172] DEPARTMENT OF HOMELAND SECURITY Coast Guard 33 CFR Part 165 [Docket No. USCG-2011-1164] RIN 1625-AA87 Security Zone; Choptank River and Cambridge Channel, Cambridge, MD..., U.S. Coast Guard, Captain of the Port Baltimore. [FR Doc. 2012-1172 Filed 1-20-12; 8:45 am]...

  1. 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…

  2. 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…

  3. Cambridge IGCSE mathematics core and extended

    CERN Document Server

    Pimentel, Ric

    2013-01-01

    The most cost effective and straightforward way to teach the revised syllabus, with all the core and extended content covered by a single book and accompanying free digital resources.  . This title has been written for the revised Cambridge IGCSE Mathematics (0580) syllabus, for first teaching from 2013.  . ·         Gives students the practice they require to deepen their understanding through plenty of questions. ·         Consolidates learning with unique digital resources on the CD, included free with every Student's Book.  . We are working with Cambridge International Examinations to gain

  4. OCR Cambridge nationals in ICT student book

    CERN Document Server

    Stuart, Sonia; Cushing, Steve

    2012-01-01

    Written by experts and in partnership with OCR, the brand-new OCR Cambridge Nationals in ICT Student's Book provides invaluable guidance for your teaching of the OCR Cambridge Nationals in ICT Level 1/2 . This textbook covers the mandatory Units 1 and 2 in detail, offering your students the knowledge and practice they require. Unit 1 - Understanding Computer Systems.; Coverage of use of applications and systems.; Case studies of how they are used for different purposes.; Exam style questions and guidance. Unit 2 - Using ICT to Create Business Solutions.; Coverage of the principles of use of re

  5. Western blot analysis of IgG responses of sheep infected with Fasciola hepatica and F.gigantica%免疫印迹分析试验感染肝片吸虫和大片吸虫绵羊的IgG应答

    Institute of Scientific and Technical Information of China (English)

    张为宇; MOREAU Emmanuelle; 黄维义; CHAUVIN Alain

    2003-01-01

    免疫印迹(Western blot)分析试验感染肝片吸虫和大片吸虫绵羊的抗片形吸虫分泌排泄产物(FhESP和FgESP)IgG应答,结果表明,FhESP中分子质量在17.1~21.6 ku之间的4条主要蛋白带仅被肝片吸虫感染以后(0WPI以后)的血清识别且不被对照组血清识别; FgESP中分子质量在19.0~71.1 ku之间的6条主要蛋白带仅被大片吸虫感染以后(0WPI以后)的血清识别且不被对照组血清识别.

  6. Weizmann ties with Cambridge in physics contest

    CERN Multimedia

    Siegel, J

    2004-01-01

    "Scientists and students from the Weizmann Institute of Science in Rehovot and Cambridge University in England have tied for first place in a physics competition aimed at simulating the future functioning of the particle accelerator being built at the European center CERN and due to open in 2007" (1/2 page)

  7. 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.

  8. 76 FR 12729 - Cambridge Environmental Inc; Transfer of Data

    Science.gov (United States)

    2011-03-08

    ... AGENCY Cambridge Environmental Inc; Transfer of Data AGENCY: Environmental Protection Agency (EPA... claimed as Confidential Business Information (CBI) by the submitter, will be transferred to Cambridge Environmental Inc. in accordance with 40 CFR 2.307(h)(3) and 2.308(i)(2). Cambridge Environmental Inc. has...

  9. Hodder Cambridge primary mathematics learner's book 5

    CERN Document Server

    King, Steph

    2017-01-01

    Endorsed by Cambridge International Examinations to support the full curriculum framework from 2011. Develop learners' mathematical fluency, problem solving and reasoning skills using the mastery approach with this series of Learner's books, aiding preparation for the Progression and Cambridge Primary Tests. - Introduces topics through engaging starter activities.- Develops mathematical language with New Words and worked examples. - Illustrates topics clearly and vividly with imaginative design and relatable characters- Builds fluency and mathematical reasoning skills by exploring, clarifying, practising and then extending concepts to ensure learners master mathematical ideas.- Enhances learners' ability to apply their skills and solve non-routine mathematical problems, by ensuring they secure a deep conceptual understanding of the subject. - Supports learners of all abilities with hints and Try this extension challenges.- Secures knowledge with problem solving integrated throughout....

  10. Cambridge Elementary students enjoy gift of computers

    Science.gov (United States)

    1999-01-01

    Children at Cambridge Elementary School, Cocoa, Fla., eagerly unwrap computer equipment donated by Kennedy Space Center. Cambridge is one of 13 Brevard County schools receiving 81 excess contractor computers thanks to an innovative educational outreach project spearheaded by the Nasa k-12 Education Services Office at ksc. Behind the children is Jim Thurston, a school volunteer and retired employee of USBI, who shared in the project. The Astronaut Memorial Foundation, a strategic partner in the effort, and several schools in rural Florida and Georgia also received refurbished computers as part of the year-long project. Ksc employees put in about 3,300 volunteer hours to transform old, excess computers into upgraded, usable units. A total of $90,000 in upgraded computer equipment is being donated.

  11. Hodder Cambridge primary mathematics learner's book 2

    CERN Document Server

    Casey, Catherine

    2017-01-01

    Endorsed by Cambridge International Examinations to support the full curriculum framework from 2011. Develop learners' mathematical fluency, problem solving and reasoning skills using the mastery approach with this series of Learner's books, aiding preparation for the Progression and Cambridge Primary Tests. - Introduces topics through engaging starter activities.- Develops mathematical language with New Words and worked examples. - Illustrates topics clearly and vividly with imaginative design and relatable characters- Builds fluency and mathematical reasoning skills by exploring, clarifying, practising and then extending concepts to ensure learners master mathematical ideas.- Enhances learners' ability to apply their skills and solve non-routine mathematical problems, by ensuring they secure a deep conceptual understanding of the subject. - Supports learners of all abilities with hints and Try this extension challenges.- Secures knowledge with problem solving integrated throughout....

  12. Hodder Cambridge primary mathematics workbook 2

    CERN Document Server

    Broadbent, Paul

    2017-01-01

    Endorsed by Cambridge International Examinations to support the full curriculum framework from 2011. Consolidate learning, deepen conceptual understanding and develop problem solving skills through practice questions; ideal for independent learning, homework or extension activities- Supports and build on knowledge gained from the Learner's Book with practice exercises.- Provides additional 'intelligent practice' through variation.- Challenges learners to deepen and extend their understanding- Incorporates the principles of variation and intelligent practice in each activity.

  13. Hodder Cambridge primary mathematics workbook 5

    CERN Document Server

    King, Steph

    2017-01-01

    Endorsed by Cambridge International Examinations to support the full curriculum framework from 2011. Consolidate learning, deepen conceptual understanding and develop problem solving skills through practice questions; ideal for independent learning, homework or extension activities- Supports and build on knowledge gained from the Learner's Book with practice exercises.- Provides additional 'intelligent practice' through variation.- Challenges learners to deepen and extend their understanding- Incorporates the principles of variation and intelligent practice in each activity.

  14. Alchemy in Cambridge. An Annotated Catalogue of Alchemical Texts and Illustrations in Cambridge Repositories.

    Science.gov (United States)

    Timmermann, Anke

    2015-01-01

    Alchemy in Cambridge captures the alchemical content of 56 manuscripts in Cambridge, in particular the libraries of Trinity College, Corpus Christi College and St John's College, the University Library and the Fitzwilliam Museum. As such, this catalogue makes visible a large number of previously unknown or obscured alchemica. While extant bibliographies, including those by M.R. James a century ago, were compiled by polymathic bibliographers for a wide audience of researchers, Alchemy in Cambridge benefits from the substantial developments in the history of alchemy, bibliography, and related scholarship in recent decades. Many texts are here identified for the first time. Another vital feature is the incorporation of information on alchemical illustrations in the manuscripts, intended to facilitate research on the visual culture of alchemy. The catalogue is aimed at historians of alchemy and science, and of high interest to manuscript scholars, historians of art and historians of college and university libraries.

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

    Science.gov (United States)

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

    2006-01-01

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

  16. The new Cambridge English course student 1

    CERN Document Server

    Swan, Michael

    1991-01-01

    The New Cambridge English Course is a course teachers and students can rely on to cover the complete range and depth of language and skills needed from beginner to upper-intermediate level. Each level is designed to provide at least 72 hours of class work using the Student's Book, with additional self-study material provided in the Practice Book. The course has a proven multi-syllabus approach which integrates work on all the vital aspects of language study: grammar, vocabulary, pronunciation, skills, notions and functions.

  17. Cambridge IGCSE english as a second language

    CERN Document Server

    Reynolds, John

    2014-01-01

    Revised edition for the 2015 syllabus offering the easiest and most cost effective way to teach both the speaking and listening components with one set of books covering two years and free digital material. This title has been written for the revised Cambridge IGCSE English as a Second Language (0510 and 0511) syllabuses, for first teaching from 2013. ? Prepares students for their exams with a focus on assessed language features, such as inference, opinion and attitude. ? Develops language abilities at an appropriate pace with extra interactive tests on a free CD-ROM. We are working with Cambr

  18. The Whipple Museum and Cavendish Laboratory, Cambridge

    Science.gov (United States)

    Pippard, Brian

    The Whipple Museum is part of the History and Philosophy of Science Department in the University of Cambridge. It is on your right as soon as you enter Free School Lane from Pembroke Street, and is normally open between 1:30 and 4:30 P.M. on weekdays. The main room, a hall with hammer-beam roof, is a relic of Stephen Perse’s school (1624) now flourishing elsewhere in the city. It houses a large collection of mathematical, physical and astronomical instruments — abaci, Napier’s bones, slide rules; sextants and other surveying instruments; telescopes, compasses and pocket sundials (especially of ivory from Nuremberg 1500-1700); and a Grand Orrery by George Adams (1750). The gallery of a second room is used for special exhibitions, often of items from the well-stocked store. Some specialist catalogues have been compiled and are on sale.

  19. Teachers Learning: Professional Development and Education. Cambridge Education Research Series

    Science.gov (United States)

    McLaughlin, Colleen, Ed.

    2012-01-01

    "Teachers Learning: Professional Development and Education" is part of The Cambridge Education Research series, edited by senior colleagues at the University of Cambridge Faculty of Education, which has a longstanding tradition of involvement in high quality, innovative teacher education and continuing professional development.…

  20. Teachers Learning: Professional Development and Education. Cambridge Education Research Series

    Science.gov (United States)

    McLaughlin, Colleen, Ed.

    2012-01-01

    "Teachers Learning: Professional Development and Education" is part of The Cambridge Education Research series, edited by senior colleagues at the University of Cambridge Faculty of Education, which has a longstanding tradition of involvement in high quality, innovative teacher education and continuing professional development.…

  1. 76 FR 13665 - Cambridge Tool & Die, Including On-Site Leased Workers From Action Total Staffing, Cambridge, OH...

    Science.gov (United States)

    2011-03-14

    ... Register on January 26, 2011 (76 FR 4731). At the request of the State agency, the Department reviewed the... Employment and Training Administration Cambridge Tool & Die, Including On-Site Leased Workers From Action Total Staffing, Cambridge, OH; Amended Certification Regarding Eligibility To Apply for...

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

    Science.gov (United States)

    Taki, Takao

    2015-01-01

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

  3. Heterogeneity of PrP in Brains of Hamsters Infected with Scrapie 263K Strain and the Dynamic Detection by Western blot%羊瘙痒病263K毒株感染的仓鼠脑中PrP分子形式多样性的动态测定

    Institute of Scientific and Technical Information of China (English)

    高建梅; 高晨; 周晓勃; 张瑾; 韩俊; 张宝云; 董小平

    2003-01-01

    正常细胞的朊蛋白(PrPC)代谢和构象的改变是引发动物和人类可传播性海绵状脑病(transmissible spongiform encephalopathies,TSEs)的根本原因.将羊瘙痒病( scrapie)仓鼠适应株263K颅内接种仓鼠, 在接种后的第20、40、50、60、70、80天, 通过Western blot 动态检测仓鼠脑中PrP存在的形式.结果在接种后第40天, 在感染动物脑组织中即检测到 PrPSc分子,比临床症状出现的时间早(平均潜伏期为66.7±1.1天),且无糖基化形式的PrP分子所占百分比在接种后期增加明显.除了标准分子量大小(30kD~35kD)的 PrP分子外,在感染动物脑中存在着高分子量和低分子量形式的PrP分子 .定量分析显示,随着接种潜伏期的延长,不同形式PrP分子的含量也在增加,其中低分子量形式的PrP分子与临床症状的出现密切相关.蛋白去糖基化实验表明,在感染动物脑组织中,除了标准分子量大小的PrP蛋白外,还存在一条更小分子量的PrP条带,而正常动物脑组织仅存在标准大小的Pr P分子.低分子量形式的PrP分子具有与全长PrP分子相类似的糖基化模式.结果提示,scrapie 263K感染的仓鼠脑组织中存在不同分子形式的PrPSc,其PrP分子的代谢可能不同于正常动物.

  4. The Evaluation of Western Blotting Method for the Diagnosis of Helicobacter Pylori Infection in Children%免疫印迹试验在诊断儿童幽门螺旋杆菌感染中的应用评价

    Institute of Scientific and Technical Information of China (English)

    陈运生; 周少明; 刘彬; 孙玲玲

    2006-01-01

    目的:评价免疫印迹试验(Western Blotting)在诊断不同年龄组及不同病理表现儿童幽门螺旋杆菌(Helicobacter pylori,HP)感染中的应用价值.方法:对450例有消化道症状的患儿,进行胃镜检查并取病变处胃黏膜组织行病理学检查,以及13C-尿素呼气试验(13C-urea breath test,13C-UBT)、血清免疫印迹试验及胃液HP-DNA -PCR检查.结果:在4岁~6岁和6岁~12岁组患儿中,13C-UBT HP检出率明显高于免疫印迹试验(P<0.01);在12岁~18岁组患儿中,13C-UBT HP检出率与免疫印迹试验比较差异无显著性(P>0.05).细胞毒性基因相关蛋白(Cytotoxin-associated gene protein)阳性的HP感染在溃疡病组明显高于慢性胃炎组和正常组(P<0.05),慢性胃炎组CagA(Cytotoxin-associated gene protein,CagA) 阳性 HP显著高与正常组(P<0.05).结论:在12岁以上的少年患儿中,免疫印迹试验可作为快速诊断HP感染的可靠手段,并且能判断出是否为产毒菌株感染.在12岁以下的儿童中,免疫印迹试验最好与13C-UBT联合使用.

  5. Recent Developments in Cambridge College Libraries

    Directory of Open Access Journals (Sweden)

    Alison Wilson

    2006-07-01

    Full Text Available Cambridge University has three tiers of libraries available to students: the University Library, departmental (subject libraries and college libraries. Over the past thirty years there has been increasing pressure on the colleges to provide more books, reader places and technical resources in their libraries, with the result that a number of new library buildings, of very different styles, have been opened. Other colleges have opted for refurbishment and extension of existing libraries. These libraries are small (30-100,000 books and intimate, often open 24 hours a day and with generous provision for lending books. Great importance is placed on keeping them at the heart of the college. Challenges for architects are the sensitive sites, restrictions on changes to listed buildings, and the limited space available. The constricted sites cause difficulties for the builders too. I will consider some solutions to these problems with reference to projects in four colleges: Pembroke, Peterhouse, Corpus Christi and Newnham. At Pembroke architects Freeland Rees Roberts have built an extension to a listed building and at Peterhouse they have adapted an adjoining room. Corpus Christi is moving its library to a Victorian building which has been internally redesigned by Wright + Wright. Newnham demolished a 1960s extension in order to develop the plot more efficiently to a design by John Miller + Partners. All the architects have shown sensitivity to the needs of their clients and ingenuity in making intensive use of limited space.

  6. The CAMbridge Emission Line Surveyor (CAMELS)

    CERN Document Server

    Thomas, C N; Maiolino, R; Goldie, D J; Acedo, E de Lera; Wagg, J; Blundell, R; Paine, S; Zeng, L

    2014-01-01

    The CAMbridge Emission Line Surveyor (CAMELS) is a pathfinder program to demonstrate on-chip spectrometry at millimetre wavelengths. CAMELS will observe at frequencies from 103-114.7 GHz, providing 512 channels with a spectral resolution of R = 3000. In this paper we describe the science goals of CAMELS, the current system level design for the instrument and the work we are doing on the detailed designs of the individual components. In addition, we will discuss our efforts to understand the impact that the design and calibration of the filter bank on astronomical performance. The shape of the filter channels, the degree of overlap and the nature of the noise all effect how well the parameters of a spectral line can be recovered. We have developed a new and rigorous method for analysing performance, based on the concept of Fisher information. This can in be turn coupled to a detailed model of the science case, allowing design trade-offs to be properly investigated.

  7. Artificial Pancreas Project at Cambridge 2013.

    Science.gov (United States)

    Hovorka, R

    2015-08-01

    The development and clinical testing of closed-loop systems (the artificial pancreas) is underpinned by advances in continuous glucose monitoring and benefits from concerted academic and industry collaborative efforts. This review describes the progress of the Artificial Pancreas Project at the University of Cambridge from 2006 to 2014. Initial studies under controlled laboratory conditions, designed to collect representative safety and performance data, were followed by short to medium free-living unsupervised outpatient studies demonstrating the safety and efficacy of closed-loop insulin delivery using a model predictive control algorithm. Accompanying investigations included assessment of the psychosocial impact and key factors affecting glucose control such as insulin kinetics and glucose absorption. Translation to other disease conditions such as critical illness and Type 2 diabetes took place. It is concluded that innovation of iteratively enhanced closed-loop systems will provide tangible means to improve outcomes and quality of life in people with Type 1 diabetes and their families in the next decade.

  8. 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

  9. CRITERIA OF POSITIVITY FOR Ig ANTIBODIES IN THE METHOD OF IMMUNE BLOTTING OF LYME DISEASE

    Directory of Open Access Journals (Sweden)

    V G Barskova

    2001-01-01

    Full Text Available There are currently no accepted criteria for positive Western blots in Russian patients with Lyme borreliosis. The purpose of the current study was to develop criteria for a positive IgG westem-blot to aid particularly in the diagnosis of patients with joint manifestation of the disorder. Patients: 97 with Lyme disease, 145 - control subjects. IgG antibody responses were determined to 3 species ofB.burgdorferi sensu lato by Western blotting, using blots prepared by manufacturer. The best discriminatory ability of test criteria was chained by requiring any 3 of 11 IgG bands, a definition that could be used with B. burgdorferi sensu stricto, B.garinii and B.afzelii strains. With these 3 antigen preparation, positive IgG blots were found in 0 to 18% of patients with localized erythema migrans of < 4 weeks duration, 23 to 39% of those with disseminated infection < 20 weeks duration, and in 39 to 46% of those with late arthritis/arthralgia of >6 months duration the specificity was 93 to 99%. Thus, IgG Western blotting may bring greater specificity to serologic testing in Lyme borreliosis, but the sensitivity is limited.

  10. The Cambridge-Cambridge ROSAT serendipity survey; 2, classification of x-ray luminous galaxies

    CERN Document Server

    Boyle, B J; Wiles, B J; Elvis, M; Boyle, B J; McMahon, R G; Wiles, B J; Elvis, M

    1995-01-01

    We present the results of an intermediate-resolution (1.5\\AA) spectroscopic study of 17 X-ray luminous narrow emission-line galaxies previously identified in the Cambridge-Cambridge {\\it ROSAT} Serendipity Survey and the {\\it Einstein} Extended Medium Sensitivity Survey. Emission-line ratios reveal that the sample is composed of ten Seyfert and seven starburst galaxies. Measured linewidths for the narrow H\\alpha emission lines lie in the range 170-460\\,km\\,s^{-1}. Five of the objects show clear evidence for asymmetry in the [OIII]\\lambda5007 emission-line profile. Broad H\\alpha emission is detected in six of the Seyfert galaxies, which range in type from Seyfert 1.5 to 2. Broad H\\beta emission is only detected in one Seyfert galaxy. The mean full width at half maximum for the broad lines in the Seyfert galaxies is {\\rm FWHM = 3900\\pm 1750\\,km\\,s^{-1}}. Broad ({\\rm FWHM = 2200\\pm 600\\,km\\,s^{-1}}) H\\alpha emission is also detected in three of the starburst galaxies, which could originate from stellar winds or ...

  11. Analyses of the Results of HIV-1 Western Blot Confirmative Test and NAT Test in Voluntary Blood Donors%无偿献血者HIV-1抗体蛋白印迹确认与核酸检测结果分析

    Institute of Scientific and Technical Information of China (English)

    吕蓉; 王伦善; 盛琪琪; 赵阳; 蒋菲菲; 李敏; 刘忠

    2012-01-01

    Objective To explore the performance of confirmative Western blot (WB) and TMA-chemical luminescence test in detecting the reactive-samples for HIV antibody ELISA assay. Methods 117 reactive specimens detected by ELISA were repeatedly tested by two methods of primary screening assay and TMA-luminescence nucleic acid amplification testing(NAT). All samples of S/CO>0. 8 were analysed by the WB confirmative test. Serological tests participated in the external quality assessment (EQA) were surported by China CDC and Australian international CITIC as well as NAT participated in the EQA of Clinical Laboratory Center of the Ministry of Health and Australian CITIC. Results The results of all 117 tested samples were as followed, in primary screening test:S/CO>1,0. 8

  12. Michele Renee Salzman, Marvina A. Sweeney & William Adler (eds., The Cambridge History of Religions in the Ancient World (2 vols. (New York: Cambridge University Press, 2013

    Directory of Open Access Journals (Sweden)

    Giorgio Baruchello

    2014-03-01

    Full Text Available Review of: Michele Renee Salzman, Marvina A. Sweeney & William Adler (eds., The Cambridge History of Religions in the Ancient World (2 vols. (New York: Cambridge University Press, 2013

  13. 8th Cambridge Workshop on Universal Access and Assistive Technology

    CERN Document Server

    Lazar, Jonathan; Heylighen, Ann; Dong, Hua

    2016-01-01

    This book presents the proceedings of the 8th Cambridge Workshop on Universal Access and Assistive Technology (CWUAAT '14), incorporating the 11th Cambridge Workshop on Rehabilitation Robotics, held in Cambridge, England in March 2016. It presents novel and state-of-the-art research from an international group of leaders in the fields of universal access and assistive technology. It explores various issues including the reconciliation of usability, accessibility and inclusive design, the design of inclusive assistive and rehabilitation systems, measuring product demand and human capabilities, data mining and visualizing inclusion, legislation in inclusive design, and situational inclusive interfaces (automotive and aerospace). This book provides an invaluable resource to researchers, postgraduates, design practitioners, therapists and clinical practitioners, as well as design teachers.

  14. Intelligence Inside the King’s College of Cambridge

    Directory of Open Access Journals (Sweden)

    Henriono Nugroho

    2008-11-01

    Full Text Available Article concerns with a stylistic analysis on a poem in terms of Systemic Functional Linguistics and Verbal Art Semiotics. The writing uses library research, qualitative data, documentary study, descriptive method and intrinsic-objective approach. The semantic analysis results in both automatized and foregrounded meanings. Then the automatized meaning produces lexical cohesion and in turn, it produces subject matter. Meanwhile, the foregrounded meaning produces the literary meaning and in turn, it creates theme. Finally, the analysis indicates that the subject matter is about the establishment of Cambridge University, the literary meaning is about eternal thoughts of Cambridge University, and the theme is about intelligence.

  15. Cambridge international AS and A level mathematics statistics

    CERN Document Server

    Porkess, Roger; Konrad, Nina

    2012-01-01

    This brand new series has been written for the University of Cambridge International Examinations course for AS and A Level Mathematics (9709). This title covers the requirements of S1 and S2. The authors are experienced examiners and teachers who have written extensively at this level, so have ensured all mathematical concepts are explained using language and terminology that is appropriate for students across the world. Students are provded with clear and detailed worked examples and questions from Cambridge International past papers, so they have the opportunity for plenty of essenti

  16. Cambridge international AS and A level mathematics mechanics

    CERN Document Server

    Porkess, Roger

    2012-01-01

    This brand new series has been written for the University of Cambridge International Examinations course for AS and A Level Mathematics (9709). This title covers the requirements of M1 and M2. The authors are experienced examiners and teachers who have written extensively at this level, so have ensured all mathematical concepts are explained using language and terminology that is appropriate for students across the world. Students are provded with clear and detailed worked examples and questions from Cambridge International past papers, so they have the opportunity for plenty of essential exam

  17. Skype'i stipendium viib Cambridge'i / Kadri Bank

    Index Scriptorium Estoniae

    Bank, Kadri

    2006-01-01

    Eesti Infotehnoloogia Sihtasutuse (EITSA) Skype'i 100 000-kroonise magistristipendiumi sai Tartu Ülikooli matemaatika-informaatikateaduskonna kevadel lõpetanud Ago-Erik Riet, kes edestas kuut konkurenti ja asub õppima Cambridge'i Ülikooli magistrantuuri matemaatikat

  18. The Cambridge Primary Review: A Reply to R. J. Campbell

    Science.gov (United States)

    Armstrong, Michael

    2010-01-01

    The author was disappointed by R. J. Campbell's sour critique of the Cambridge Primary Review in "FORUM" Volume 52 Number 1 2010. His description of the Review's proposals on curriculum and pedagogy as "backward-looking, cumbersome and partial" is such a bizarre misjudgement that it calls for some response. The author comments in turn on R. J.…

  19. Skype'i stipendium viib Cambridge'i / Kadri Bank

    Index Scriptorium Estoniae

    Bank, Kadri

    2006-01-01

    Eesti Infotehnoloogia Sihtasutuse (EITSA) Skype'i 100 000-kroonise magistristipendiumi sai Tartu Ülikooli matemaatika-informaatikateaduskonna kevadel lõpetanud Ago-Erik Riet, kes edestas kuut konkurenti ja asub õppima Cambridge'i Ülikooli magistrantuuri matemaatikat

  20. The Cambridge Primary Review: A Reply to R. J. Campbell

    Science.gov (United States)

    Armstrong, Michael

    2010-01-01

    The author was disappointed by R. J. Campbell's sour critique of the Cambridge Primary Review in "FORUM" Volume 52 Number 1 2010. His description of the Review's proposals on curriculum and pedagogy as "backward-looking, cumbersome and partial" is such a bizarre misjudgement that it calls for some response. The author comments in turn on R. J.…

  1. Group photograph: Dark Energy Survey Autumn Collaboration meeting Cambridge 2016 12 December 2016 - 16 December 2016

    OpenAIRE

    Smith, Amanda

    2016-01-01

    Group photograph of participants in the Dark Energy Survey Autumn collaboration meeting Cambridge 2016 12 December 2016 - 16 December 2016 taken outside the Cambridge Observatory building on 14 December 2016, by Amanda Smith, Graphics Officer.

  2. 78 FR 52802 - Manufacturer of Controlled Substances; Notice of Application; Cambridge Isotope Lab

    Science.gov (United States)

    2013-08-26

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF JUSTICE Drug Enforcement Administration Manufacturer of Controlled Substances; Notice of Application; Cambridge Isotope Lab... 01, 2013, Cambridge Isotope Lab, 50 Frontage Road, Andover, Massachusetts 01810, made application...

  3. 77 FR 38086 - Manufacturer of Controlled Substances, Notice of Application, Cambridge Isotope Lab

    Science.gov (United States)

    2012-06-26

    ... From the Federal Register Online via the Government Publishing Office DEPARTMENT OF JUSTICE Drug Enforcement Administration Manufacturer of Controlled Substances, Notice of Application, Cambridge Isotope Lab... 7, 2012, Cambridge Isotope Lab, 50 Frontage Road, Andover, Massachusetts 01810, made application...

  4. 40 CFR 81.205 - Zanesville-Cambridge Intrastate Air Quality Control Region.

    Science.gov (United States)

    2010-07-01

    ... 40 Protection of Environment 17 2010-07-01 2010-07-01 false Zanesville-Cambridge Intrastate Air... Air Quality Control Regions § 81.205 Zanesville-Cambridge Intrastate Air Quality Control Region. The Zanesville-Cambridge Intrastate Air Quality Control Region (Ohio) consists of the territorial...

  5. University of Cambridge deploys Procket Networks' PRO/8801

    CERN Multimedia

    2003-01-01

    Procket Networks, a provider of high performance Internet Protocol (IP) technology and products has announced that the University of Cambridge has deployed the PRO/8801(TM) router into its research network to develop industry-leading deep packet inspection applications. The major application for this deployment is to identify and understand new traffic patterns created by large scale scientific computations and downloads such as the GRID (1 page).

  6. Metaphor Analysis of Chinese Premier Wen’s Cambridge Speech

    Institute of Scientific and Technical Information of China (English)

    LUO Luo

    2014-01-01

    Metaphor is more than an ostensible decoration of language. It is an integral part of human thought of ideologized world. This article analyzes the metaphor use of Chinese Premier Wen Jiabao’s speech at Cambridge in February 2009, in an at-tempt to display how the preferred metaphors serve the purpose of this speech and reflect Premier Wen ’s construction of Chi-na’s situation.

  7. Traffic Sign Recognition System based on Cambridge Correlator Image Comparator

    OpenAIRE

    J. Turan; L. Ovsenik; T. Harasthy

    2012-01-01

    Paper presents basic information about application of Optical Correlator (OC), specifically Cambridge Correlator, in system to recognize of traffic sign. Traffic Sign Recognition System consists of three main blocks, Preprocessing, Optical Correlator and Traffic Sign Identification. The Region of Interest (ROI) is defined and chosen in preprocessing block and then goes to Optical Correlator, where is compared with database of Traffic Sign. Output of Optical Correlation is correlation plane, w...

  8. 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.

  9. 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.

  10. Cambridge IGCSE and international certificate French foreign language

    CERN Document Server

    Grime, Yvette; Thacker, Mike

    2013-01-01

    This brand-new Student Book provides a grammar-led approach with extensive exam preparation that will help you develop independent, culturally aware students of French ready for the exam. The book is written to the latest Cambridge International Examinations syllabus by experienced teachers. Extensive use of French reflects the style of the exams and, with specific advice and practice, it helps students use the acquired skills to their best ability. Topics on Francophone cultures are integrated throughout to ensure students gain the cultural awareness that is at the heart of this qualification

  11. WORKING WITH ARCHIVES: CAMBRIDGE ECONOMICS THROUGH THE MAGNIFYING GLASS

    Directory of Open Access Journals (Sweden)

    Maria Cristina Marcuzzo

    2012-12-01

    Full Text Available Research in economics using documentary archives provides clearer interpretations about the ideas and their development throughout time, in the context of the writing process in relation to the interlocutors and antagonists, and the nature of the problems addressed. This document presents examples of works with tough drafts, correspondence, tables of contents, notes, and related material of four economists of the "Cambridge Group": Piero Sraffa, Richard Kahn, Joan Robinson, and John Maynard Keynes. In each case the findings and importance of the research done in the files are described.

  12. The Cambridge MRI database for animal models of Huntington disease.

    Science.gov (United States)

    Sawiak, Stephen J; Morton, A Jennifer

    2016-01-01

    We describe the Cambridge animal brain magnetic resonance imaging repository comprising 400 datasets to date from mouse models of Huntington disease. The data include raw images as well as segmented grey and white matter images with maps of cortical thickness. All images and phenotypic data for each subject are freely-available without restriction from (http://www.dspace.cam.ac.uk/handle/1810/243361/). Software and anatomical population templates optimised for animal brain analysis with MRI are also available from this site.

  13. Skin Regeneration Symposium Cambridge, 12-13 April 2016.

    Science.gov (United States)

    Hill, Rosalind

    2016-07-01

    The Annual Skin Regeneration Symposium, held in Cambridge, UK, 12-13 April 2016, explored the latest advancements in skin repair, regeneration and restoration, and the impact this has on patients. With over 140 delegates from the disciplines of burn and trauma care, chronic wounds and esthetic medicine, the symposium sparked lively debate and the sharing of results from interesting case studies, clinical trials and basic research to support the use of a Regenerative Epithelial Suspension produced using the ReCell(®) technology. Furthermore, it enabled delegates and speakers alike to share ideas and discuss how to improve the quality of care for patients.

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

    Science.gov (United States)

    Lee, Der-Yen; Chang, Geen-Dong

    2015-01-01

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

  15. Lipid-binding analysis using a fat blot assay.

    NARCIS (Netherlands)

    Munnik, T.; Wierzchowiecka, M.

    2013-01-01

    Protein-lipid interactions play an important role in lipid metabolism, membrane trafficking and cell -signaling by regulating protein localization, activation, and function. The Fat Blot assay is a relatively simple and inexpensive method to examine these interactions using nitrocellulose

  16. Mixed lubrication after rewetting of blotted pleural mesothelium.

    Science.gov (United States)

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

    2013-01-15

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

  17. The Cambridge Structural Database in retrospect and prospect.

    Science.gov (United States)

    Groom, Colin R; Allen, Frank H

    2014-01-13

    The Cambridge Crystallographic Data Centre (CCDC) was established in 1965 to record numerical, chemical and bibliographic data relating to published organic and metal-organic crystal structures. The Cambridge Structural Database (CSD) now stores data for nearly 700,000 structures and is a comprehensive and fully retrospective historical archive of small-molecule crystallography. Nearly 40,000 new structures are added each year. As X-ray crystallography celebrates its centenary as a subject, and the CCDC approaches its own 50th year, this article traces the origins of the CCDC as a publicly funded organization and its onward development into a self-financing charitable institution. Principally, however, we describe the growth of the CSD and its extensive associated software system, and summarize its impact and value as a basis for research in structural chemistry, materials science and the life sciences, including drug discovery and drug development. Finally, the article considers the CCDC's funding model in relation to open access and open data paradigms.

  18. Applications of the Cambridge Structural Database in chemical education.

    Science.gov (United States)

    Battle, Gary M; Ferrence, Gregory M; Allen, Frank H

    2010-10-01

    The Cambridge Structural Database (CSD) is a vast and ever growing compendium of accurate three-dimensional structures that has massive chemical diversity across organic and metal-organic compounds. For these reasons, the CSD is finding significant uses in chemical education, and these applications are reviewed. As part of the teaching initiative of the Cambridge Crystallographic Data Centre (CCDC), a teaching subset of more than 500 CSD structures has been created that illustrate key chemical concepts, and a number of teaching modules have been devised that make use of this subset in a teaching environment. All of this material is freely available from the CCDC website, and the subset can be freely viewed and interrogated using WebCSD, an internet application for searching and displaying CSD information content. In some cases, however, the complete CSD System is required for specific educational applications, and some examples of these more extensive teaching modules are also discussed. The educational value of visualizing real three-dimensional structures, and of handling real experimental results, is stressed throughout.

  19. [Clinical usefulness and psychometric properties of the Cambridge Behavioural Inventory].

    Science.gov (United States)

    Fernandez-Martinez, R; Kokoulina, E; Carballido-Araujo, E; Garcia-Fuertes, I; Gutierrez-Martinez, O; Santiago-Lopez, F; Vazquez-Batan, P

    2016-11-16

    Introduccion. Un area importante de la evaluacion neuropsicologica son los sintomas psicologicos y conductuales. El inventario conductual de CambridgeCambridge Behavioural Inventory (CBI)– es una medida de autoinforme dirigida a allegados que recoge una amplia variedad de sintomas conductuales que pueden darse en el curso de las enfermedades neurologicas. El principal objetivo del estudio es comprobar la utilidad clinica de su adaptacion al castellano. Sujetos y metodos. El CBI fue cumplimentado por 215 allegados de pacientes remitidos desde los servicios de neurologia y psiquiatria. Se compararon los perfiles del CBI de cuatro grupos de pacientes formados sobre la base de sus principales caracteristicas clinicas, datos psicometricos, pruebas de imagen y juicio clinico del profesional solicitante del estudio neuropsicologico. Resultados. La mayoria de las escalas (10 de 13) del CBI tuvo valores de consistencia interna aceptables, y las escalas de memoria y atencion/orientacion, correlaciones elevadas con medidas objetivas de memoria y orientacion temporal. Los perfiles del CBI de los grupos de pacientes con distintas condiciones (trastorno organico de la memoria, trastorno funcional de la memoria, variante conductual de demencia frontotemporal y enfermedad de Alzheimer) fueron consistentes con sus principales caracteristicas. Conclusiones. El CBI es un instrumento psicometricamente fiable y con adecuada validez convergente y discriminante que puede ser util en el proceso de evaluacion neuropsicologica, aportando informacion relevante no solo sobre el funcionamiento cognitivo y las capacidades funcionales, sino tambien sobre los sintomas conductuales y psicologicos de los pacientes con trastornos cognitivos.

  20. Applications of the Cambridge Structural Database in chemical education1

    Science.gov (United States)

    Battle, Gary M.; Ferrence, Gregory M.; Allen, Frank H.

    2010-01-01

    The Cambridge Structural Database (CSD) is a vast and ever growing compendium of accurate three-dimensional structures that has massive chemical diversity across organic and metal–organic compounds. For these reasons, the CSD is finding significant uses in chemical education, and these applications are reviewed. As part of the teaching initiative of the Cambridge Crystallographic Data Centre (CCDC), a teaching subset of more than 500 CSD structures has been created that illustrate key chemical concepts, and a number of teaching modules have been devised that make use of this subset in a teaching environment. All of this material is freely available from the CCDC website, and the subset can be freely viewed and interrogated using WebCSD, an internet application for searching and displaying CSD information content. In some cases, however, the complete CSD System is required for specific educational applications, and some examples of these more extensive teaching modules are also discussed. The educational value of visualizing real three-dimensional structures, and of handling real experimental results, is stressed throughout. PMID:20877495

  1. Sediment remediation of the Hespeler Mill Pond, Cambridge, Ontario

    Energy Technology Data Exchange (ETDEWEB)

    Angeloni, D.; Eby, M.; Jarvis, S.; Martin, P. [Univ. of Guelph, School of Engineering, Guelph, Ontario (Canada)]. E-mail: danielle.angeloni@earthtech.ca

    2002-06-15

    'Full text:' Low dissolved oxygen levels and large accumulated sediment remediation alternatives were examined to assemble the Hespeler Mill Pond, Cambridge (HMP) into a healthier and more desirable recreational area in the City of Cambridge. The theory that a large amount of sediment has been deposited into the HMP from the Speed River upstream over a number of years predicts the depressed oxygen levels, high nutrient-loading rates and the odour problems in the summer months. The initial phase in the remediation plan for this project involved extensive background research and investigation. The focus was on determining the characteristics of the sediment and the history of the pond, to ultimately decide if the sediment was the source of the issues. Dissolved oxygen field tests and sediment sampling were conducted to get information on the magnitude of the problem and the environmental hazards potentially present in the pond. The pond was modelled utilising the Streeter-Phelps oxygen-sag model to predict the oxygen deficit. Biochemical Oxygen Demand (BOD{sub 5}) testing was completed to determine the oxygen demand in the pond. These tests were conducted by using water samples obtained from various sample points at the pond. The proposed solution is a combined dredging and aeration approach. Mechanical dredging using a clamshell bucket and the installation of aerators is expected to solve the dissolved oxygen and water quality issues. (author)

  2. From Cape Town to Cambridge: Orthopaedic trauma in contrasting environments.

    Science.gov (United States)

    Lawrence, John E; Khanduja, Vikas

    2016-05-18

    To compare the trauma experience gained by a trainee at a United Kingdom major trauma centre and a secondary level hospital in South Africa. A profile of inpatient trauma cases during a five-week period in Addenbrooke's Hospital, Cambridge and Somerset Hospital, Cape Town was created. This was achieved by recording various parameters for each patient admitted including age, gender, injury, mechanism of injury and postal/area code. This, together with details of the departments themselves, allows a comparison of the amount and variety of orthopaedic trauma cases experienced by an individual trainee in each setting. The trauma profiles differed significantly. Patients in Cape Town were younger and more likely to be male. In the young, injury in Cape Town was more likely to occur due to assault or being struck by a vehicle, whilst patients in Cambridge were more likely to be injured whilst in a vehicle or in high energy falls. In older patients, trauma at both centres was almost exclusively due to mechanical falls. In a given age group, injuries at the two centres were similar, however the majority of patients admitted to Addenbrooke's were elderly, resulting in less variation in the overall injury profile. The trauma profile of a major trauma centre in the United Kingdom is less varied than that of a South African secondary centre, with significantly fewer cases per surgeon. This suggests a more varied training experience in the developing world with a greater caseload.

  3. The Cambridge Equation with government activity revisited A Equação de Cambridge com atividade governamental revisitada

    Directory of Open Access Journals (Sweden)

    Ricardo Silva Azevedo Araujo

    2010-12-01

    Full Text Available This paper offers an analysis of the steady-state distributional features found in a Kaldor-Pasinetti process, in which the government sector is allowed to run persistent deficits that may be financed through different instruments. Productive capital and bonds generate single rates of return, while workers' saving propensity remains uniform. This paper seeks to establish a generalization of Cambridge Eauqtion, considering the specific contributions of Steedman (1972, Pasinetti (1989, Dalziel (1991, and Faria (2000.Neste artigo faz-se uma análise das características distributivas do processo Kaldor-Pasinetti, assumindo-se que o setor governamental incorre em persistentes déficits que podem ser financiados através de diferentes instrumentos, como a emissão de títulos e de moeda. Através dessa abordagem é possível estudar como a atividade governamental afeta a distribuição de renda entre capitalistas e trabalhadores e assim obter generalizações do Teorema de Cambridge em que versões anteriores como as de Steedman (1972, Pasinetti (1989, Dalziel (1991 e Faria (2000 surgem como casos particulares.

  4. Hodder Cambridge primary mathematics teacher's resource pack 5

    CERN Document Server

    King, Steph

    2017-01-01

    Endorsed by Cambridge International Examinations to support the full curriculum framework from 2011. Save time and ensure full coverage of learning objectives with concise teaching notes and assessment support, built on a mastery approach to embed mathematical understanding. - Ensures clarity about the mastery approach with clear background information- Saves time with suggestions for starter activities relating to activities from the Learner's book, to introduce and explore new concepts- Assists formative and summative assessment with essential guidance- Allows flexibility and freedom to adapt ideas to the needs of the class whilst still being supportive and informative- Provides answers to Workbook questions, practice tests and Learner's book activities- Safeguards understanding with further activities and ideas for teaching to ensure that all learners master the concepts required.

  5. 7th Cambridge Workshops on Universal Access and Assistive Technology

    CERN Document Server

    Lazar, J; Heylighen, A; Dong, H; Inclusive Designing : Joining Usability, Accessibility, and Inclusion

    2014-01-01

    ‘Inclusive Designing’ presents the proceedings of the seventh Cambridge Workshop on Universal Access and Assistive Technology (CWUAAT '14). It represents a unique multi-disciplinary workshop for the Inclusive Design Research community where designers, computer scientists, engineers, architects, ergonomists, policymakers and user communities can exchange ideas. The research presented at CWUAAT '14 develops methods, technologies, tools and guidance that support product designers and architects to design for the widest possible population for a given range of capabilities, within a contemporary social and economic context. In the context of developing demographic changes leading to greater numbers of older people and people with disabilities, the general field of Inclusive Design Research strives to relate the capabilities of the population to the design of products. Inclusive populations of older people contain a greater variation in sensory, cognitive and physical user capabilities. These variations may be...

  6. Traffic Sign Recognition System based on Cambridge Correlator Image Comparator

    Directory of Open Access Journals (Sweden)

    J. Turan

    2012-06-01

    Full Text Available Paper presents basic information about application of Optical Correlator (OC, specifically Cambridge Correlator, in system to recognize of traffic sign. Traffic Sign Recognition System consists of three main blocks, Preprocessing, Optical Correlator and Traffic Sign Identification. The Region of Interest (ROI is defined and chosen in preprocessing block and then goes to Optical Correlator, where is compared with database of Traffic Sign. Output of Optical Correlation is correlation plane, which consist of highly localized intensities, know as correlation peaks. The intensity of spots provides a measure of similarity and position of spots, how images (traffic signs are relatively aligned in the input scene. Several experiments have been done with proposed system and results and conclusion are discussed.

  7. Hodder Cambridge primary mathematics teacher's resource pack 2

    CERN Document Server

    Fernandes, Sarah-Anne

    2017-01-01

    Endorsed by Cambridge International Examinations to support the full curriculum framework from 2011. Save time and ensure full coverage of learning objectives with concise teaching notes and assessment support, built on a mastery approach to embed mathematical understanding. - Ensures clarity about the mastery approach with clear background information- Saves time with suggestions for starter activities relating to activities from the Learner's book, to introduce and explore new concepts- Assists formative and summative assessment with essential guidance- Allows flexibility and freedom to adapt ideas to the needs of the class whilst still being supportive and informative- Provides answers to Workbook questions, practice tests and Learner's book activities- Safeguards understanding with further activities and ideas for teaching to ensure that all learners master the concepts required.

  8. A Comparison between Oxford University and Cambridge University

    Institute of Scientific and Technical Information of China (English)

    MEI Qiong-hui; ZHENG Yun-yan; YANG Zhi-min

    2016-01-01

    牛津大学和剑桥大学是两所世界著名的英国大学。他们不同于英国的其他大学,但两者本身却在历史、组织构造、教学风格方面拥有很多的相似之处。与此同时,它们在发展过程、框架结构、教学重点方面也有着不同之处。牛津大学和剑桥大学是两所世界一流的大学,它们因此成为世界各地的年青人所向往的理想大学。这两所大学将继续保持它们在将来高等教育界的显著地位。%Oxford University and Cambridge University in the United Kingdom are two world-famous universities, they differ from other universities in the United Kingdom, but they themselves have much in common in history, organizational makeup, and teaching style. Meanwhile, they also have some differences in development, framework, and educational focus. As two of the first-class universities in the world, Oxford University and Cambridge University have become the ideal universities that many young people all over the world yearn to enter. The two universities will keep themselves in the foreground of higher education in the future.

  9. Anneli Randla kaitses doktorikraadi Cambridge'is / Anneli Randla ; interv. Reet Varblane

    Index Scriptorium Estoniae

    Randla, Anneli

    1999-01-01

    5. mail kaitses Cambridge'is esimese eesti kunstiteadlasena doktorikraadi Anneli Randla. Töö teema: kerjusmungaordukloostrite arhitektuur Põhja-Euroopas. Juhendaja dr. Deborah Howard. Doktorikraadile esitatavatest nõudmistest, doktoritöö kaitsmisest, magistrikraadi kaitsnu õppimisvõimalustest Cambridge's.

  10. In search of low cost titanium: the fray farthing chen (FFC) cambridge process

    CSIR Research Space (South Africa)

    Oosthuizen, SJ

    2011-03-01

    Full Text Available This article explores the Fray Farthing Chen (FFC) Cambridge process, a novel method for the electrochemical deoxidation of metal oxides in molten salt, discovered at the University of Cambridge in 1997. The process was hailed as a highly promising...

  11. In search of low cost titanium: the Fray Farthing Chen (FFC) Cambridge process

    CSIR Research Space (South Africa)

    Oosthuizen, SJ

    2010-10-01

    Full Text Available This article explores the Fray Farthing Chen (FFC) Cambridge Process, a novel method for the electrochemical de-oxidation of metal oxides in molten salt, discovered at the University of Cambridge in 1997. The process was hailed as a highly promising...

  12. Investigating the Impact of Cambridge International Assessments on U.S. Stakeholders: Student and Teacher Perceptions

    Science.gov (United States)

    Shaw, Stuart

    2011-01-01

    As part of the continuing program to study the impact of its international assessments, the University of Cambridge International Examinations ("Cambridge") has undertaken a series of studies investigating the impact on a range of US stakeholders. This paper reports on research designed to respond to a series of washback and impact…

  13. Assessing the Impact of Arts and Humanities Research at the University of Cambridge. Technical Report

    Science.gov (United States)

    Levitt, Ruth; Celia, Claire; Diepeveen, Stephanie; Chonaill, Siobhan Ni; Rabinovich, Lila; Tiessen, Jan

    2010-01-01

    This project for the University of Cambridge and the Arts and Humanities Research Council (AHRC) assesses the impacts of arts and humanities research at the University of Cambridge. Evidence from interviews, a survey of research staff and detailed case studies indicates that these disciplines already have a broad range of impacts. Many of these…

  14. A snuff, Sir? Et ego in Arcadia - op sabbatical in Cambridge

    NARCIS (Netherlands)

    Hondius, E.H.

    2005-01-01

    Een maand lang heb ik in Cambridge onderzoek mogen doen. In deze column wil ik daar verslag van uitbrengen. Niet van mijn onderzoek — ik heb vooral rondgekeken in de bibliotheek en gesproken met collega's om te zien wat momenteel in de common law te koop is - maar van Cambridge zelf.

  15. Success in the US: Are Cambridge International Assessments Good Preparation for University Study?

    Science.gov (United States)

    Shaw, Stuart; Bailey, Clare

    2011-01-01

    This article focuses on the research being conducted by University of Cambridge International Examinations (Cambridge) to ensure that its international assessments prepare students as well as Advanced Placement and International Baccalaureate for continued studies in colleges and universities. The primary purpose of the research is to highlight…

  16. Investigating the Impact of Cambridge International Assessments on U.S. Stakeholders: Student and Teacher Perceptions

    Science.gov (United States)

    Shaw, Stuart

    2011-01-01

    As part of the continuing program to study the impact of its international assessments, the University of Cambridge International Examinations ("Cambridge") has undertaken a series of studies investigating the impact on a range of US stakeholders. This paper reports on research designed to respond to a series of washback and impact questions…

  17. 77 FR 64143 - Manufacturer of Controlled Substances; Notice of Registration; Cambridge Isotope Lab

    Science.gov (United States)

    2012-10-18

    ... Enforcement Administration Manufacturer of Controlled Substances; Notice of Registration; Cambridge Isotope Lab By Notice dated June 18, 2012, and published in the Federal Register on June 26, 2012, 77 FR 38086, Cambridge Isotope Lab, 50 Frontage Road, Andover, Massachusetts 01810, made application by renewal to...

  18. Assessing the Impact of Arts and Humanities Research at the University of Cambridge. Technical Report

    Science.gov (United States)

    Levitt, Ruth; Celia, Claire; Diepeveen, Stephanie; Chonaill, Siobhan Ni; Rabinovich, Lila; Tiessen, Jan

    2010-01-01

    This project for the University of Cambridge and the Arts and Humanities Research Council (AHRC) assesses the impacts of arts and humanities research at the University of Cambridge. Evidence from interviews, a survey of research staff and detailed case studies indicates that these disciplines already have a broad range of impacts. Many of these…

  19. 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

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

    Science.gov (United States)

    2004-02-14

    adding hydroxylapatite (0.5% w/v; Calbiochem fast flow; Calbiochem, San Diego , CA). The suspensions were shaken on ice for 1–2 h, then incubated...Pellizzari, C. Recchi, G. Napolitani , M. Mock, C. Montecucco, Anthrax lethal factor cleaves the N-terminus of MAPKKs and induces tyrosine/threonine

  1. 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.

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

    Science.gov (United States)

    1992-09-16

    antibodies to retroviral proteins in parIents with primary Sjogren’s syndrome. Arthritis and Rheumatism 1990; 33:774-781. 51. Kunhl P, Seidl S, Holzberger...erythematosus, rheumatoid arthritis , juvenile onset diabetes, thyroiditis, and Crohn’s disease), were less likely to be tested electively for HIV, and a lower...association if both the IWB and autoantibodies were caused by another factor. Parity could reflect alloimmunization during pregnancy , producing

  3. The Cambridge-Cambridge x-ray serendipity survey. 2: Classification of x-ray luminous galaxies

    Science.gov (United States)

    Boyle, B. J.; Mcmahon, R. G.; Wilkes, B. J.; Elvis, Martin

    1994-01-01

    We present the results of an intermediate-resolution (1.5 A) spectroscopic study of 17 x-ray luminous narrow emission-line galaxies previously identified in the Cambridge-Cambridge ROSAT Serendipity Survey and the Einstein Extended Medium Sensitivity Survey. Emission-line ratios reveal that the sample is composed of ten Seyfert and seven starburst galaxies. Measured linewidths for the narrow H alpha emission lines lie in the range 170 - 460 km s(exp -1). Five of the objects show clear evidence for asymmetry in the (OIII) lambda 5007 emission-line profile. Broad H alpha emission is detected in six of the Seyfert galaxies, which range in type from Seyfert 1.5 to 2. Broad H beta emission is only detected in one Seyfert galaxy. The mean full width at half maximum for the broad lines in the Seyfert galaxies is FWHM = 3900 +/- 1750 km s(exp -1). Broad (FWHM = 2200 +/- 600 km s(exp -1) H alpha emission is also detected in three of the starburst galaxies, which could originate from stellar winds or supernovae remnants. The mean Balmer decrement for the sample is H alpha / H beta = 3, consistent with little or no reddening for the bulk of the sample. There is no evidence for any trend with x-ray luminosity in the ratio of starburst galaxies to Seyfert galaxies. Based on our previous observations, it is therefore likely that both classes of object comprise approximately 10 percent of the 2 keV x-ray background.

  4. Findings from the Harvard Medical School Cambridge Integrated Clerkship, a Year-Long Longitudinal Psychiatry Experience.

    Science.gov (United States)

    Cheng, Elisa; Hirsh, David; Gaufberg, Elizabeth; Griswold, Todd; Wesley Boyd, J

    2017-06-23

    The Harvard Medical School Cambridge Integrated Clerkship is a longitudinal integrated clerkship that has provided an alternative clinical model for medical education in psychiatry since its inception in 2004. This study was undertaken in an effort to better understand the student experience of the Cambridge Integrated Clerkship and how it may have impacted students' perceptions of and interest in psychiatry, as well as performance. Qualitative surveys were sent via e-mail to the first 11 student cohorts who had completed the Cambridge Integrated Clerkship (from 2004 to 2014) and for whom we had e-mail addresses (N = 100), and the free-text responses were coded thematically. All available standardized scoring data and residency match data for Cambridge Integrated Clerkship graduates were obtained. From 2006 to 2014, 12 out of 73 Cambridge Integrated Clerkship students who entered the match chose a psychiatry residency (16.4%), four times more than students in traditional clerkships at Harvard Medical School (3.8% of 1355 students) or the national average (4.1% of 146,066 US applicants). Thirty of the 100 surveyed Cambridge Integrated Clerkship graduates (30%) responded to the qualitative survey with free-text remarks on a number of themes. Cambridge Integrated Clerkship students compared positively to their classmates in terms of standardized test performance. Their fourfold higher match rate into psychiatry compared to other students raises intriguing questions as to what role a longitudinal clerkship might have played in developing interest in psychiatry as a career.

  5. Fingerprinting of Natural Product by Eastern Blotting Using Monoclonal Antibodies

    Directory of Open Access Journals (Sweden)

    Hiroyuki Tanaka

    2012-01-01

    Full Text Available We succeeded in developing the fingerprint of natural product by eastern blotting using monoclonal antibodies. After developing and separating them on a TLC plate, solasodine glycosides are oxidized by NaIO4 and reacted with a protein to give conjugates which are recognized with anti-solamargine monoclonal antibody (MAb. Anti-solamargine MAb having wide cross-reactivity can stain and detect all solasodine glycosides by fingerprint. Different sensitivity between solamargine and solasonine was observed. The detection limit was 1.6 ng of solasonine. The hydrolysed products of solamargine were determined by fingerprint of eastern blotting compared to their Rf values depending on the sugar number. Fingerprint by eastern blotting using anti-ginsenoside Rb1 MAb distinguished the formula containing ginseng prescribed in traditional Chinese medicine. By double-staining of ginsenosides it is possible to suggest that the staining color shows the pharmacological activity, such as the purple bands indicate ginsenosides having stimulation activity, and the blue color indicated compound like ginsenosides possessed the depression affect for the central nervous system (CNS, respectively.

  6. Factor V Leiden, factor V Cambridge, factor II GA20210, and methylenetetrahydrofolate reductase in cerebral venous and sinus thrombosis: A case-control study.

    Science.gov (United States)

    Saadatnia, Mohammad; Salehi, Mansour; Movahedian, Ahmad; Shariat, Seyed Ziaeddin Samsam; Salari, Mehri; Tajmirriahi, Marzieh; Asadimobarakeh, Elham; Salehi, Rasoul; Amini, Gilda; Ebrahimi, Homa; Kheradmand, Ehsan

    2015-06-01

    Factor V G1691A (FV Leiden), FII GA20210, and methylenetetrahydrofolate reductase (MTHFR) C677T mutations are the most common genetic risk factors for thromboembolism in the Western countries. However, there is rare data in Iran about cerebral venous and sinus thrombosis (CVST) patients. The aim of this study was to evaluate the frequency of common genetic thrombophilic factors in CVST patients. Forty consequently CVST patients from two University Hospital in Isfahan University of Medical Sciences aged more than 15 years from January 2009 to January 2011 were recruited. In parallel, 51 healthy subjects with the same age and race from similar population selected as controls. FV Leiden, FII GA20210, MTHFR C677T, and FV Cambridge gene mutations by polymerase chain reaction technique were evaluated in case and control groups. FV Leiden, FII GA20210, and FV Cambridge gene mutations had very low prevalence in both case (5%, 2%, 0%) and control (2.5%, 0%, 0%) and were not found any significant difference between groups. MTHFR C677T mutations was in 22 (55%) of patients in case group and 18 (35.5%) of control group (P = 0.09). This study showed that the prevalence of FV Leiden, FII GA20210, and FV Cambridge were low. Laboratory investigations of these mutations as a routine test for all patients with CVST may not be cost benefit.

  7. Worries of Pregnant Women: Testing the Farsi Cambridge Worry Scale

    Directory of Open Access Journals (Sweden)

    Forough Mortazavi

    2016-01-01

    Full Text Available Pregnancy adds many sources of concerns to women’s daily life worries. Excessive worry can affect maternal physiological and psychological state that influences the pregnancy outcomes. The aim of this study was to validate the Cambridge Worry Scale (CWS in a sample of Iranian pregnant women. After translation of the CWS, ten experts evaluated the items and added six items to the 17-item scale. In a descriptive cross-sectional study, 405 of pregnant women booked for prenatal care completed the Farsi CWS. We split the sample randomly. Exploratory factor analysis (EFA was conducted on the first half of the sample to disclose the factorial structure of the 23-item scale. The results of the EFA on the Farsi CWS indicated four factors altogether explained 51.5% of variances. Confirmatory factor analysis (CFA was done on the second half of the sample. The results of the CFA showed that the model fit our data (chi-square/df = 2.02, RMSEA = 0.071, SRMR = 0.071, CFI = 0.95, and NNFI = 0.94. Cronbach’s alpha coefficient for the Farsi CWS was 0.883. The Farsi CWS is a reliable and valid instrument for understanding common pregnancy worries in the third trimester of pregnancy in Iranian women.

  8. The hermeneutics of mental symptoms in the Cambridge School

    Directory of Open Access Journals (Sweden)

    Massimiliano Aragona

    2015-12-01

    Full Text Available Current Psychiatry is in crisis. Decades of neuroscientific research have not yet delivered adequate explanations or treatments. One reason for this failure may be the wrongness of its central assumption, namely that mental symptoms and disorders are natural kinds. The Cambridge School has proposed that a new Epistemology must be constructed for Psychiatry, and that this should start with the development of a new model of mental symptom-formation. ‘Mental symptoms’ should be considered as hermeneutic co-constructions occurring in a intersubjective space created by the dialogue between sufferer and healer. Subjective experiences (caused either by neurobiological or psychosocial upheaval penetrate the awareness of sufferers causing perplexity and/or distress. To understand, handle and communicate these experiences, sufferers proceed to configure them by means of templates borrowed from their own culture. Importantly, however, the same neurobiological information can be configured into different symptoms; and different neurobiological information into the same symptom. Therefore, ‘mental symptoms’ are dissimilar hybrid combinations of neurobiological and cultural information. To be ethical, therapeutic interventions must take into account such dissimilarities. Blind manipulation of the brain in all cases should be considered as counterproductive.

  9. Worries of Pregnant Women: Testing the Farsi Cambridge Worry Scale.

    Science.gov (United States)

    Mortazavi, Forough; Akaberi, Arash

    2016-01-01

    Pregnancy adds many sources of concerns to women's daily life worries. Excessive worry can affect maternal physiological and psychological state that influences the pregnancy outcomes. The aim of this study was to validate the Cambridge Worry Scale (CWS) in a sample of Iranian pregnant women. After translation of the CWS, ten experts evaluated the items and added six items to the 17-item scale. In a descriptive cross-sectional study, 405 of pregnant women booked for prenatal care completed the Farsi CWS. We split the sample randomly. Exploratory factor analysis (EFA) was conducted on the first half of the sample to disclose the factorial structure of the 23-item scale. The results of the EFA on the Farsi CWS indicated four factors altogether explained 51.5% of variances. Confirmatory factor analysis (CFA) was done on the second half of the sample. The results of the CFA showed that the model fit our data (chi-square/df = 2.02, RMSEA = 0.071, SRMR = 0.071, CFI = 0.95, and NNFI = 0.94). Cronbach's alpha coefficient for the Farsi CWS was 0.883. The Farsi CWS is a reliable and valid instrument for understanding common pregnancy worries in the third trimester of pregnancy in Iranian women.

  10. Who Needs a Revision? 20 Years of Cambridge Shunt Lab.

    Science.gov (United States)

    Czosnyka, Zofia; Czosnyka, Marek; Pickard, John D; Chari, Aswin

    2016-01-01

    Shunt testing independent of manufacturers provides knowledge that can significantly improve the management of patients with hydrocephalus. The Cambridge Shunt Evaluation Laboratory was created 20 years ago. Thanks to financial support from the Department of Health (1993-1998), all shunts in use in the UK were systematically evaluated, with "blue reports" being published. Later new devices were tested as they appeared in public domain.Twenty-six models have been evaluated. The majority of the valves had a non-physiologically low hydrodynamic resistance that may result in over-drainage, both related to posture and during nocturnal cerebral vasogenic waves. A long distal catheter increases the resistance of these valves by 100-200 %. Drainage through valves without a siphon-preventing mechanism is very sensitive to body posture. Shunts with siphon-preventing accessories offer a reasonable resistance to negative outlet pressure. Bench parameters were used to test shunt performance in vivo using infusion tests. A criterion for correctly performing a shunt procedure was established. Pressure measured in the shunt prechamber during the plateau phase of infusion should not remain more than 5 mmHg above the le shunt's operating pressure plus hydrodynamic resistance of the valve multiplied by the infusion rate. "Critical levels" for every shunt and every performance level have been used in the shunt testing wizard of ICM+ software.

  11. Development and current status of the "Cambridge" loudness models.

    Science.gov (United States)

    Moore, Brian C J

    2014-10-13

    This article reviews the evolution of a series of models of loudness developed in Cambridge, UK. The first model, applicable to stationary sounds, was based on modifications of the model developed by Zwicker, including the introduction of a filter to allow for the effects of transfer of sound through the outer and middle ear prior to the calculation of an excitation pattern, and changes in the way that the excitation pattern was calculated. Later, modifications were introduced to the assumed middle-ear transfer function and to the way that specific loudness was calculated from excitation level. These modifications led to a finite calculated loudness at absolute threshold, which made it possible to predict accurately the absolute thresholds of broadband and narrowband sounds, based on the assumption that the absolute threshold corresponds to a fixed small loudness. The model was also modified to give predictions of partial loudness-the loudness of one sound in the presence of another. This allowed predictions of masked thresholds based on the assumption that the masked threshold corresponds to a fixed small partial loudness. Versions of the model for time-varying sounds were developed, which allowed prediction of the masked threshold of any sound in a background of any other sound. More recent extensions incorporate binaural processing to account for the summation of loudness across ears. In parallel, versions of the model for predicting loudness for hearing-impaired ears have been developed and have been applied to the development of methods for fitting multichannel compression hearing aids.

  12. 1919: psychology and psychoanalysis, Cambridge and London - Myers, Jones and MacCurdy.

    Science.gov (United States)

    Forrester, John

    2008-01-01

    Viewing the reception of psychoanalysis in Britain from Cambridge, the paper examines the intertwining histories of the post-War British Psychological Society and the founding of the British Psycho-Analytical Society, following the initiatives of the two principal psychological entrepreneurs of the era, Charles Myers and Ernest Jones, Reforms in Cambridge in which psychoanalysis played a significant part are analysed, including the foundation of a Clinic for Nervous Diseases and the establishment of a separate Department of Experimental Psychology. The career of J. T. MacCurdy, Jones's student and Lecturer in Psychopathology at Cambridge, is discussed.

  13. 75 FR 38128 - Sensata Technologies MA, Inc., Power Controls Division, Formerly Known As Airpax Corp., Cambridge...

    Science.gov (United States)

    2010-07-01

    ... Employment and Training Administration Sensata Technologies MA, Inc., Power Controls Division, Formerly Known..., 2010, applicable to workers of Sansata Technologies MA, Incorporated, Power Controls Division, formerly... under the control of the Cambridge, Maryland location of Sensata Technologies MA, Incorporated,...

  14. Cambridge Polytrauma Pathway: Are we making appropriately guided decisions?

    Science.gov (United States)

    Wynell-Mayow, William; Guevel, Borna; Quansah, Benjamin; O'Leary, Ronan; Carrothers, Andrew D

    2016-10-01

    Addenbrooke's Hospital, the Major Trauma Centre for the East of England Trauma Network, received 1070 major trauma patients between 1st January and 31st December 2014. In order to improve care, an audit was performed of 59 patients meeting our own selection criteria for orthopaedic polytrauma between 1st January 2013 and 31st December 2013. The Cambridge Polytrauma Pathway was devised through NCEPOD guidelines, literature review, internal and external discussion. It facilitates provision of best practice Early Appropriate Care, encompassing - multidisciplinary consultant decisions around the patient in our Neurological and Trauma Critical Care Unit, early full body trauma CT scans, serial measurements of lactate and fibrinogen levels, and out-of-hours orthopaedic theatre reserved for life-and-limb threatening injuries. Re-audit was conducted of 15 patients meeting selection criteria, admitted between 1st October 2014 and 31st March 2015. Significant improvements in recording of lactate and fibrinogen were demonstrated, both on admission (lactate - p<0.000, fibrinogen - p=0.015), and preoperatively (lactate - p=0.003, fibrinogen - p=0.030). Time to trauma CT was unchanged (p=0.536) with a median time to CT of 0.53h at re-audit (IQR 0.48-0.75). The number of patients receiving definitive orthopaedic intervention out-of-hours reduced from 8 to zero (p=0.195). The approach of facilitating management decisions to be made at early daytime MDT meetings has been adopted. It is anticipated that this pathway will improve outcomes in orthopaedic polytrauma patients and it is recommended that either the GOS-E, or the EQ-5D scoring systems be introduced to assess this. Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.

  15. Plastination in Anatomy Learning: An Experience at Cambridge University.

    Science.gov (United States)

    Latorre, Rafael; Bainbridge, David; Tavernor, Angie; López Albors, Octavio

    2016-01-01

    Due to lack of objective data, the benefits of using plastination in combination with wet dissection in teaching gross anatomy are unknown. The aim of this study was to obtain objective evidence from students regarding the effectiveness of combining plastinated specimens (PS) with an established gross anatomy education program at Cambridge University that uses wet cadaver dissection and small-group tutorials. For a complete academic year, a total of 135 PS were used alongside wet cadaver dissections. The PS were also available for small-group tutorials. An anonymous closed questionnaire, using a 5-point numerical-estimation Likert scale, was used to gather information relating to the effectiveness of the PS. The level of student satisfaction with the combined use of wet dissections and PS was high, although higher (p<.05) for second-year students (98.4%) than for first-year students (95.5%). Students felt the specimens allowed them to see details that were often more difficult to identify in their dissections, for instance nerves. Voluntary use of PS was higher (p<.01) for second-year students (96.9%), who had previously experienced anatomy teaching with cadaver dissection alone, than for first-year students (77.7%). Overall, 97.7% of all students thought that the PS helped them understand and learn anatomy. All students surveyed (100%) recommended the use of PS in the future. Students considered the use of PS in the dissection room combined with wet cadaver dissection to be beneficial when learning anatomy, particularly when combined with their use during small-group tutorials.

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

    Science.gov (United States)

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

    2007-04-15

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

  17. 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.

  18. Brief history of the Cambridge STEM aberration correction project and its progeny

    Energy Technology Data Exchange (ETDEWEB)

    Brown, L. Michael [Cavendish Laboratory, University of Cambridge, J.J. Thomson Avenue, Cambridge CB3 0HE (United Kingdom); Batson, Philip E. [Institute for Advanced Materials, Devices and Nanotechnology, Rutgers University, Piscataway, NJ 08854 (United States); Department of Physics, Rutgers University, Piscataway, NJ 08854 (United States); Department of Materials Science, Rutgers University, Piscataway, NJ 08854 (United States); Dellby, Niklas [Nion Company, 11515 NE 118th Street, Kirkland, WA 98034 (United States); Krivanek, Ondrej L. [Nion Company, 11515 NE 118th Street, Kirkland, WA 98034 (United States); Department of Physics, Arizona State University, Tempe, AZ 85287 (United States)

    2015-10-15

    We provide a brief history of the project to correct the spherical aberration of the scanning transmission electron microscope (STEM) that started in Cambridge (UK) and continued in Kirkland (WA, USA), Yorktown Heights (NY, USA), and other places. We describe the project in the full context of other aberration correction research and related work, partly in response to the incomplete context presented in the paper “In quest of perfection in electron optics: A biographical sketch of Harald Rose on the occasion of his 80th birthday”, recently published in Ultramicroscopy. - Highlights: • We provide a brief history of the Cambridge project to correct the spherical aberration of the scanning transmission electron microscope (STEM). • We describe the project in the full context of other aberration correction work and related research. • We summarize our corrector development work that followed the Cambridge project, and which was the first to reach higher spatial resolution than any non-corrected electron microscope.

  19. Taxonomic review of the New World spider genus Elaver O. Pickard-Cambridge, 1898 (Araneae, Clubionidae).

    Science.gov (United States)

    Saturnino, Regiane; Bonaldo, Alexandre Bragio

    2015-11-23

    Elaver O. Pickard-Cambridge, 1898 is characterized and redescribed, including 49 species occurring from the United States to Argentina. Thirty seven previously known species are redescribed: Elaver achuca (Roddy, 1966) revalidated, E. balboae (Chickering, 1937), E. barroana (Chickering, 1937), E. calcarata (Kraus, 1955), E. carlota (Bryant, 1940), E. chisosa (Roddy, 1966), E. crinophora (Franganillo, 1934), E. crocota (O. Pickard-Cambridge, 1896), E. albicans (Franganillo, 1930) name restored, E. depuncta O. Pickard-Cambridge, 1898, E. elaver (Bryant, 1940), E. excepta (L. Koch, 1866), E. grandivulva (Mello-Leitão, 1930), E. hortoni (Chickering, 1937), E. implicata (Gertsch, 1941), E. juana (Bryant, 1940), E. kohlsi (Gertsch & Jellison, 1939), E. linguata (F.O. Pickard-Cambridge, 1900), E. madera (Roddy, 1966), E. mirabilis (O. Pickard-Cambridge, 1896) new. comb., E. mulaiki (Gertsch, 1935), E. multinotata (Chickering, 1937), E. orvillei (Chickering, 1937), E. placida O. Pickard-Cambridge, 1898, E. portoricensis (Petrunkevitch, 1930), E. quadrata (Kraus, 1955), E. richardi (Gertsch, 1941), E. sericea O. Pickard-Cambridge, 1898, E. sigillata (Petrunkevitch, 1925), E. simplex (O. Pickard-Cambridge, 1896), E. texana (Gertsch, 1933), E. tigrina O. Pickard-Cambridge, 1898 name restored, E. tricuspis (F.O. Pickard-Cambridge, 1900), E. tristani (Banks, 1909), E. tumivulva (Banks, 1909), E. valvula (F.O. Pickard-Cambridge, 1900) and E. wheeleri (Roewer, 1933). Ten new species are described: E. candelaria n. sp. and E. helenae n. sp. from Mexico; E. arawakan n. sp. from Haiti; E. lizae n. sp. from Costa Rica; E. darwichi n. sp. from Ecuador; E. juruti n. sp., E. tourinhoae n. sp. and E. vieirae n. sp. from Brazil; E. shinguito n. sp. from Peru and E. beni n. sp. from Bolivia. The female of E. hortoni is described for the first time. Lectotypes are designated for E. sigillata and its actual female is described for the first time. Four new synonyms are proposed: E. languida

  20. Water quality in the Cambridge, Massachusetts, drinking-water source area, 2005-8

    Science.gov (United States)

    Smith, Kirk P.; Waldron, Marcus C.

    2015-01-01

    During 2005-8, the U.S. Geological Survey, in cooperation with the Cambridge, Massachusetts, Water Department, measured concentrations of sodium and chloride, plant nutrients, commonly used pesticides, and caffeine in base-flow and stormwater samples collected from 11 tributaries in the Cambridge drinking-water source area. These data were used to characterize current water-quality conditions, to establish a baseline for future comparisons, and to describe trends in surface-water quality. The data also were used to assess the effects of watershed characteristics on surface-water quality and to inform future watershed management.

  1. Analysis and Assessment of Cambridge English Skills Real Reading 2 within the Scope of Communicative Approach

    Institute of Scientific and Technical Information of China (English)

    张润霞

    2015-01-01

    this essay analyzes and assesses Cambridge English Skills Real Reading 2 from the perspective of communicative approach.The first part analyzes the theoretical basis of the textbook,including the organizational,pragmatic competence.The second part analyzes the syllabus contents.The last part analyzes the material and tasks of it.In summary,Cambridge English Skills Real Reading 2 is mainly communicative textbook from several aspects based on the detailed assessment and explanation.It can be helpful to the designing of reading textbooks.

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

    Science.gov (United States)

    Colgan, D J

    1993-01-01

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

  3. Curriculum, Pedagogy, and the Cambridge Primary Review: A Response to R. J. Campbell

    Science.gov (United States)

    Armstrong, Michael

    2011-01-01

    This article presents the author's response to R.J. Campbell's critique of the "Cambridge Primary Review," which was published in the autumn of 2009. The author argues that Campbell's description of the "Review's" central proposals on curriculum and pedagogy as "backward-looking and inadequately theorised" is so misjudged as to call for a…

  4. Psychiatry in the Harvard Medical School-Cambridge Integrated Clerkship: An Innovative, Year-Long Program

    Science.gov (United States)

    Griswold, Todd; Bullock, Christopher; Gaufberg, Elizabeth; Albanese, Mark; Bonilla, Pedro; Dvorak, Ramona; Epelbaum, Claudia; Givon, Lior; Kueppenbender, Karsten; Joseph, Robert; Boyd, J. Wesley; Shtasel, Derri

    2012-01-01

    Objective: The authors present what is to their knowledge the first description of a model for longitudinal third-year medical student psychiatry education. Method: A longitudinal, integrated psychiatric curriculum was developed, implemented, and sustained within the Harvard Medical School-Cambridge Integrated Clerkship. Curriculum elements…

  5. Measuring Impairments in Memory and Executive Function in Older People Using the Revised Cambridge Cognitive Examination

    NARCIS (Netherlands)

    Kessels, R.P.C.; Mimpen, G.; Melis, R.J.F.; Olde Rikkert, M.G.M.

    2009-01-01

    Objectives: The Revised Cambridge Cognitive Examination (CAMCOG-R) is a cognitive screen that has been used to discriminate individuals with dementia from cognitively intact older people. It consists of items assessing various cognitive domains, but the construct validity of the cognitive subscores

  6. Measuring impairments in memory and executive function in older people using the Revised Cambridge Cognitive Examination.

    NARCIS (Netherlands)

    Kessels, R.P.C.; Mimpen, G.; Melis, R.J.F.; Olde Rikkert, M.G.M.

    2009-01-01

    OBJECTIVES: The Revised Cambridge Cognitive Examination (CAMCOG-R) is a cognitive screen that has been used to discriminate individuals with dementia from cognitively intact older people. It consists of items assessing various cognitive domains, but the construct validity of the cognitive subscores

  7. Up the Garden Path: A Chemical Trail through the Cambridge University Botanic Garden

    Science.gov (United States)

    Battle, Gary M.; Kyd, Gwenda O.; Groom, Colin R.; Allen, Frank H.; Day, Juliet; Upson, Timothy

    2012-01-01

    The living world is a rich source of chemicals with many medicines, dyes, flavorings, and foodstuffs having their origins in compounds produced by plants. We describe a chemical trail through the plant holdings of the Cambridge University Botanic Gardens. Visitors to the gardens are provided with a laminated trail guide with 22 stopping points…

  8. The Target of the Question: A Taxonomy of Textual Features for Cambridge University "O" Levels English

    Science.gov (United States)

    Benjamin, Shanti Isabelle

    2015-01-01

    This study investigates the typical textual features that are most frequently targeted in short-answer reading comprehension questions of the Cambridge University "O" Level English Paper 2. Test writers' awareness of how textual features impact on understanding of meanings in text decisions will determine to great extent their decisions…

  9. The Singapore-Cambridge General Certificate of Education Advanced-Level General Paper Examination

    Science.gov (United States)

    Hassan, Nurul Huda; Shih, Chih-Min

    2013-01-01

    This article describes and reviews the Singapore-Cambridge General Certificate of Education Advanced Level General Paper (GP) examination. As a written test that is administered to preuniversity students, the GP examination is internationally recognised and accepted by universities and employers as proof of English competence. In this article, the…

  10. Legacies, Policies and Prospects: One Year on from the Cambridge Primary Review

    Science.gov (United States)

    Alexander, Robin

    2011-01-01

    This article features the "Cambridge Primary Review." The "Review" has been supported from the beginning by Esmee Fairbairn Foundation, and this has given it the independence which is essential to its credibility. Its remit was to investigate, report and make recommendations on the condition and future of primary education in…

  11. Up the Garden Path: A Chemical Trail through the Cambridge University Botanic Garden

    Science.gov (United States)

    Battle, Gary M.; Kyd, Gwenda O.; Groom, Colin R.; Allen, Frank H.; Day, Juliet; Upson, Timothy

    2012-01-01

    The living world is a rich source of chemicals with many medicines, dyes, flavorings, and foodstuffs having their origins in compounds produced by plants. We describe a chemical trail through the plant holdings of the Cambridge University Botanic Gardens. Visitors to the gardens are provided with a laminated trail guide with 22 stopping points…

  12. Curriculum, Pedagogy, and the Cambridge Primary Review: A Response to R. J. Campbell

    Science.gov (United States)

    Armstrong, Michael

    2011-01-01

    This article presents the author's response to R.J. Campbell's critique of the "Cambridge Primary Review," which was published in the autumn of 2009. The author argues that Campbell's description of the "Review's" central proposals on curriculum and pedagogy as "backward-looking and inadequately theorised" is so misjudged as to call for a…

  13. The Target of the Question: A Taxonomy of Textual Features for Cambridge University "O" Levels English

    Science.gov (United States)

    Benjamin, Shanti Isabelle

    2015-01-01

    This study investigates the typical textual features that are most frequently targeted in short-answer reading comprehension questions of the Cambridge University "O" Level English Paper 2. Test writers' awareness of how textual features impact on understanding of meanings in text decisions will determine to great extent their decisions…

  14. A Comparison of the Abilities Measured by the Cambridge and Educational Testing Service EFL Test Batteries.

    Science.gov (United States)

    Bachman, Lyle F.; And Others

    1990-01-01

    The abilities measured by the First Certificate of English (FCE) administered by the Cambridge Local Examinations Syndicate are compared with the Test of English as a Foreign Language (TOEFL) administered by the Educational Testing Service. The analyses suggest that the FCE and TOEFL appear to measure the same common aspect of language…

  15. What To Look for in ESL Admission Tests: Cambridge Certificate Exams, IELTS, and TOEFL.

    Science.gov (United States)

    Chalhoub-Deville, Micheline; Turner, Carolyn E.

    2000-01-01

    Familiarizes test users with issues to consider when employing assessments for screening and admission purposes. Examines the purpose, content, and scoring methods of three English-as-a-Second-Language admissions tests--the Cambridge certificate exams, International English Language Teaching System, and Test of English as a Foreign…

  16. Ideology, Class and Rationality: A Critique of Cambridge International Examinations' "Thinking Skills" Curriculum

    Science.gov (United States)

    Lim, Leonel

    2012-01-01

    This article undertakes a critique of the aims and objectives of "Thinking Skills", one of the most widely and internationally used curricula in the teaching of thinking, offered by the University of Cambridge International Examinations. By engaging in a critical discourse analysis of how political and class biases are (re-)produced in the forms…

  17. Legacies, Policies and Prospects: One Year on from the Cambridge Primary Review

    Science.gov (United States)

    Alexander, Robin

    2011-01-01

    This article features the "Cambridge Primary Review." The "Review" has been supported from the beginning by Esmee Fairbairn Foundation, and this has given it the independence which is essential to its credibility. Its remit was to investigate, report and make recommendations on the condition and future of primary education in England. Its scope…

  18. The Singapore-Cambridge General Certificate of Education Advanced-Level General Paper Examination

    Science.gov (United States)

    Hassan, Nurul Huda; Shih, Chih-Min

    2013-01-01

    This article describes and reviews the Singapore-Cambridge General Certificate of Education Advanced Level General Paper (GP) examination. As a written test that is administered to preuniversity students, the GP examination is internationally recognised and accepted by universities and employers as proof of English competence. In this article, the…

  19. Linguistic Turn and Gendering Language in the Cambridge Advanced Learner's Dictionary

    Science.gov (United States)

    Arimbi, Diah A.; Kwary, Deny A.

    2016-01-01

    Language constructs how humans perceive things. Since language is a human construction, it tends to be biased as it is mainly men's construction. Using gender perspectives, this paper attempts to discuss the imbalance in gender representations found in the examples given in an English learner's dictionary, that is, the "Cambridge Advanced…

  20. Mime, Music and Drama on the Eighteenth-Century Stage. The Ballet d'Action. Edward Nye, Cambridge-New York, Cambridge University Press, 2011

    Directory of Open Access Journals (Sweden)

    Stefania Onesti

    2012-11-01

    Full Text Available Mime, Music and Drama on the Eighteenth-Century Stage by Edward Nye (Cambridge University Press, 2011 has the merit of inspiring a strong reflection on ballet d'action, connected with cultural, literaturary and philosophic environment of Eighteenth century. The author, with brilliant insight and careful historical research, explores the most debated issues of the new genre, providing an unusual interpretation. The review traces the focal points and the structure of the book, developing further consideration of some of the most challenging aspects offered by the text.

  1. An Improved Cambridge Filter Pad Extraction Methodology to Obtain More Accurate Water and “Tar” Values: In Situ Cambridge Filter Pad Extraction Methodology

    Directory of Open Access Journals (Sweden)

    Ghosh David

    2014-07-01

    Full Text Available Previous investigations by others and internal investigations at Philip Morris International (PMI have shown that the standard trapping and extraction procedure used for conventional cigarettes, defined in the International Standard ISO 4387 (Cigarettes -- Determination of total and nicotine-free dry particulate matter using a routine analytical smoking machine, is not suitable for high-water content aerosols. Errors occur because of water losses during the opening of the Cambridge filter pad holder to remove the filter pad as well as during the manual handling of the filter pad, and because the commercially available filter pad holder, which is constructed out of plastic, may adsorb water. This results in inaccurate values for the water content, and erroneous and overestimated values for Nicotine Free Dry Particulate Matter (NFDPM. A modified 44 mm Cambridge filter pad holder and extraction equipment which supports in situ extraction methodology has been developed and tested. The principle of the in situ extraction methodology is to avoid any of the above mentioned water losses by extracting the loaded filter pad while kept in the Cambridge filter pad holder which is hermetically sealed by two caps. This is achieved by flushing the extraction solvent numerous times through the hermetically sealed Cambridge filter pad holder by means of an in situ extractor. The in situ methodology showed a significantly more complete water recovery, resulting in more accurate NFDPM values for high-water content aerosols compared to the standard ISO methodology. The work presented in this publication demonstrates that the in situ extraction methodology applies to a wider range of smoking products and smoking regimens, whereas the standard ISO methodology only applies to a limited range of smoking products and smoking regimens, e.g., conventional cigarettes smoked under ISO smoking regimen. In cases where a comparison of yields between the PMI HTP and

  2. Western Sufism

    DEFF Research Database (Denmark)

    Sedgwick, Mark

    Western Sufism is sometimes dismissed as a relatively recent "new age" phenomenon, but in this book, Mark Sedgwick argues that it actually has very deep roots, both in the Muslim world and in the West. In fact, although the first significant Western Sufi organization was not established until 1915......, the first Western discussion of Sufism was printed in 1480, and Western interest in some of the ideas that are central to Sufi thought goes back to the thirteenth century. Sedgwick starts with the earliest origins of Western Sufism in late antique Neoplatonism and early Arab philosophy, and traces later...

  3. Antithrombin Cambridge II(A384S) mutation frequency and antithrombin activity levels in 120 of deep venous thrombosis and 150 of cerebral infarction patients in a single center in Southern China.

    Science.gov (United States)

    Zhang, Guang-sen; Tang, Yang-ming; Tang, Mei-qing; Qing, Zi-Ju; Shu, Chang; Tang, Xiang-qi; Deng, Ming-yang; Tan, Li-ming

    2010-09-01

    Antithrombin Cambridge II(A384S) mutation shows a relatively high frequency in western population. Some studies suggest that the mutation is an independent genetic risk factor both for deep vein thrombosis (DVT) and for arterial thrombosis, but whether the mutation has racial difference or has a general significance for thrombophilia remains unclear. In this study we performed an analysis of the prevalence of the mutation in Chinese southern population; Also, the antithrombin activity levels were evaluated in each investigated individual. The studies included 120 patients with DVT, 150 patients with cerebral infarction, and 110 controls. The mutation was detected using polymerase chain reaction/PvuII restrictive fragment length polymorphism procedures. Antithrombin activity assay was done using chromogenic substrate method. The results showed that no antithrombin Cambridge II mutation was detected in all three groups (DVT, cerebral infarction and controls), the incidence was 0/380. Plasma antithrombin activity was 91.37% +/- 16.15% in the DVT patients and 102.68% +/- 13.10% in the controls; the antithrombin activity was significantly reduced in the DVT group (P Cambridge II mutation has a racial difference, and may not be a valuable risk factor of thrombophilia in Asian population, and antithrombin deficiency remains a major genetic risk factor for DVT patients in China.

  4. The Cambridge Car Memory Test: a task matched in format to the Cambridge Face Memory Test, with norms, reliability, sex differences, dissociations from face memory, and expertise effects.

    Science.gov (United States)

    Dennett, Hugh W; McKone, Elinor; Tavashmi, Raka; Hall, Ashleigh; Pidcock, Madeleine; Edwards, Mark; Duchaine, Bradley

    2012-06-01

    Many research questions require a within-class object recognition task matched for general cognitive requirements with a face recognition task. If the object task also has high internal reliability, it can improve accuracy and power in group analyses (e.g., mean inversion effects for faces vs. objects), individual-difference studies (e.g., correlations between certain perceptual abilities and face/object recognition), and case studies in neuropsychology (e.g., whether a prosopagnosic shows a face-specific or object-general deficit). Here, we present such a task. Our Cambridge Car Memory Test (CCMT) was matched in format to the established Cambridge Face Memory Test, requiring recognition of exemplars across view and lighting change. We tested 153 young adults (93 female). Results showed high reliability (Cronbach's alpha = .84) and a range of scores suitable both for normal-range individual-difference studies and, potentially, for diagnosis of impairment. The mean for males was much higher than the mean for females. We demonstrate independence between face memory and car memory (dissociation based on sex, plus a modest correlation between the two), including where participants have high relative expertise with cars. We also show that expertise with real car makes and models of the era used in the test significantly predicts CCMT performance. Surprisingly, however, regression analyses imply that there is an effect of sex per se on the CCMT that is not attributable to a stereotypical male advantage in car expertise.

  5. RNase protection assays and RNA gel blots: a direct comparison of sensitivity.

    Science.gov (United States)

    Higgs, D C; Colbert, J T

    1992-01-01

    RNase protection assays are commonly thought to be a more sensitive means of detecting and quantitating specific mRNAs than are RNA gel blots (Northern blots). We have directly compared the sensitivity of these two approaches by assaying for known amounts of in vitro synthesized beta-glucuronidase mRNA. With the probes and protocols employed here, the ability to detect a specific mRNA was similar whether RNase protection or RNA gel blot analyses were performed.

  6. PET/MRI in the infarcted mouse heart with the Cambridge split magnet

    Energy Technology Data Exchange (ETDEWEB)

    Buonincontri, Guido, E-mail: gb396@cam.ac.uk [Wolfson Brain Imaging Centre, University of Cambridge, Box 65, Addenbrooke' s Hospital, Hills Road, Cambridge, CB2 0QQ (United Kingdom); Sawiak, Stephen J. [Wolfson Brain Imaging Centre, University of Cambridge, Box 65, Addenbrooke' s Hospital, Hills Road, Cambridge, CB2 0QQ (United Kingdom); Behavioural and Clinical Neuroscience Institute, University of Cambridge, Cambridge (United Kingdom); Methner, Carmen; Krieg, Thomas [Department of Medicine, University of Cambridge, Cambridge (United Kingdom); Hawkes, Robert C.; Adrian Carpenter, T. [Wolfson Brain Imaging Centre, University of Cambridge, Box 65, Addenbrooke' s Hospital, Hills Road, Cambridge, CB2 0QQ (United Kingdom)

    2013-02-21

    Chronic heart failure, as a result of acute myocardial infarction, is a leading cause of death worldwide. Combining diagnostic imaging modalities may aid the direct assessment of experimental treatments targeting heart failure in vivo. Here we present preliminary data using the Cambridge combined PET/MRI imaging system in a mouse model of acute myocardial infarction. The split-magnet design can deliver uncompromised MRI and PET performance, for better assessment of disease and treatment in a preclinical environment.

  7. Nutrition in medical education: reflections from an initiative at the University of Cambridge

    OpenAIRE

    Ball L; Crowley J; Laur C; Rajput-Ray M; Gillam S; Ray S

    2014-01-01

    Lauren Ball,1 Jennifer Crowley,2 Celia Laur,3 Minha Rajput-Ray,3 Stephen Gillam,4 Sumantra Ray3 1Nutrition and Dietetics, School of Allied Health Sciences, Centre for Health Practice Innovation, Griffith University, Queensland, Brisbane, Australia; 2Faculty of Medical and Health Sciences, University of Auckland, Auckland, New Zealand; 3Need for Nutrition Education/Innovation Programme, Medical Research Council Human Nutrition Research, Cambridge, UK; 4Department of Public Health and Primary C...

  8. Sir Joseph Barcroft, Cambridge placental and fetal research (1933-1966) and inter-generational Science.

    Science.gov (United States)

    Boyd, Robert; Boyd, C A Richard

    2010-01-01

    The nature of Cambridge (UK) placental and fetal research in the middle third of the twentieth century is reviewed on the basis of published literature and personal recollection. Joseph Barcroft is a central figure who came to fetal research late in an extremely productive career which is briefly sketched. Contemporaneous Cambridge academics in the field included J.D. Boyd (the authors father), J. Hammond, F.H.A. Marshall, R.A. McCance, J. Needham, A.S. Parkes and Elsie Widdowson. The then current Cambridge academic geography is explained and features of its scientific life such as funding, institutional structure and ethos, teaching and clinical duties, domestic and gender roles, and political context, including war and empire, are briefly considered. The testing of research findings against general principles and use of quantitative thinking are identified as important features. Intergenerational connections, often within individual families, are identified as a striking feature. The long-term impact of Cambridge work of this period; locally, in current trophoblast and feto-placental genetic research, in Oxford in probably influencing G.S. Dawes research leadership, and internationally, especially through D.H. Barron, and through him to the Denver School, is considered. That human placental and embryological specimens collected by J.D. Boyd have received a new lease of life as the "Boyd Collection", including use by Allen Enders is noted. Mechanisms for the maintenance of scientific quality and productivity during the period, mainly through the scientist himself relying on an internalised sense of "obligation", are contrasted with those current in the UK and more widely; formal peer-review at frequent intervals, with subsequent allocation of short-term funding. The strengths and weaknesses of each are considered.

  9. Inequality and growth in neo-Kaleckian and Cambridge growth theory

    OpenAIRE

    Thomas I. Palley

    2016-01-01

    This paper examines the relationship between inequality and growth in the neo-Kaleckian and Cambridge growth models. The paper explores the channels whereby functional and personal income distribution impact growth. The growth - inequality relationship can be negative or positive, depending on the economy's characteristics. Contrary to widespread claims, inequality per se does not impact growth through macroeconomic channels. Instead, both growth and inequality are impacted by changes in the ...

  10. Nutrition in medical education: reflections from an initiative at the University of Cambridge.

    Science.gov (United States)

    Ball, Lauren; Crowley, Jennifer; Laur, Celia; Rajput-Ray, Minha; Gillam, Stephen; Ray, Sumantra

    2014-01-01

    Landmark reports have confirmed that it is within the core responsibilities of doctors to address nutrition in patient care. There are ongoing concerns that doctors receive insufficient nutrition education during medical training. This paper provides an overview of a medical nutrition education initiative at the University of Cambridge, School of Clinical Medicine, including 1) the approach to medical nutrition education, 2) evaluation of the medical nutrition education initiative, and 3) areas identified for future improvement. The initiative utilizes a vertical, spiral approach during the clinically focused years of the Cambridge undergraduate and graduate medical degrees. It is facilitated by the Nutrition Education Review Group, a group associated with the UK Need for Nutrition Education/Innovation Programme, and informed by the experiences of their previous nutrition education interventions. Three factors were identified as contributing to the success of the nutrition education initiative including the leadership and advocacy skills of the nutrition academic team, the variety of teaching modes, and the multidisciplinary approach to teaching. Opportunities for continuing improvement to the medical nutrition education initiative included a review of evaluation tools, inclusion of nutrition in assessment items, and further alignment of the Cambridge curriculum with the recommended UK medical nutrition education curriculum. This paper is intended to inform other institutions in ongoing efforts in medical nutrition education.

  11. Oxford and Cambridge Boat Race: Performance, Pacing and Tactics Between 1890 and 2014.

    Science.gov (United States)

    Edwards, Andrew M; Guy, Joshua H; Hettinga, Florentina J

    2016-10-01

    Currently no studies have examined the historical performances of Oxford and Cambridge Boat Race crews in the context of performance, pacing and tactics which is surprising as the event has routinely taken place annually for over 150 years on the same course. The purpose of this study was twofold, to firstly examine the historical development of performances and physical characteristics of crews over 124 years of the Oxford and Cambridge Boat Race between 1890 and 2014 and secondly to investigate the pacing and tactics employed by crews over that period. Linear regression modelling was applied to investigate the development of performance and body size for crews of eight male individuals over time from Boat Race archive data. Performance change over time was further assessed in 10-year clusters while four intra-race checkpoints were used to examine pacing and tactics. Significant correlations were observed between performance and time (1890-2014) for both Oxford (r = -0.67; p Cambridge (r = -0.64; p Cambridge (1168 ± 89.8 s) during 1890-2014. Crew performance times improved over time with significant gains from baseline achieved in the 1950s (Cambridge) and the 1960s (Oxford), which coincided with significant change in the physicality of the competing crews (p < 0.01). There was no tactical advantage from commencing on either the Surrey or Middlesex station beyond chance alone; however, all crews (n = 228) adopted a fast-start strategy, with 81 % of victories achieved by the crew leading the race at the first intra-race checkpoint (24 % of total distance). Crews leading the race at the final checkpoint (83 % of total distance; 1143 m) achieved victory on 94 % of occasions. Performances and physical characteristics of the crews have changed markedly since 1890, with faster heavier crews now common. Tactically, gaining the early lead position with a fast-start strategy seems particularly meaningful to success in the Boat Race throughout the years

  12. 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...

  13. [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.

  14. 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.

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

    Science.gov (United States)

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

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

    OpenAIRE

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

    2015-01-01

    This report provides data that are specifically related to the differential sialylation of nutrient deprived breast cancer cells to sialic acid supplementation in support of the research article entitled, “Nutrient-deprived cancer cells preferentially use sialic acid to maintain cell surface glycosylation" [1]. Particularly, breast cancer cells, when supplemented with sialic acid under nutrient deprivation, display sialylated glycans at the cell surface, but non-malignant mammary cells show s...

  17. Characterization of Polyclonal Antibody Induced by Autoantibody TPO (Thyroidperoxidase) From Autoimmune Thyroid Disease (AITD) Serum with ELISA and Western Blotting

    OpenAIRE

    Maulidya Aulia Fiqriyana; Aulanni'am Aulanni'am; Anna Roosdiana

    2013-01-01

    Autoantibody TPO is a potential marker for early detection of autoimmune thyroid disease (AITD). Autoantibody TPO has a specifity and a sensitivity ranging from 82% to100% in comparison to other AITD serology markers. Concentration of autoantibody TPO in sera had a positive correlation with activities of chronic AITD. This research have been conducted to investigate the characteristic of polyclonal antibody TPO induced by autoantibody TPO from serum of AITD patients. The autoantibody TPO was ...

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

    Science.gov (United States)

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

    2014-06-01

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

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

    Science.gov (United States)

    Klinz, F J

    1994-10-01

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

  20. Innovations and challenges in renal cell carcinoma: summary statement from the Second Cambridge Conference.

    Science.gov (United States)

    Atkins, Michael B; Ernstoff, Marc S; Figlin, Robert A; Flaherty, Keith T; George, Daniel J; Kaelin, William G; Kwon, Eugene D; Libermann, Towia A; Linehan, W Marston; McDermott, David F; Ochoa, Augusto C; Pantuck, Allan J; Rini, Brian I; Rosen, Mark A; Sosman, Jeffrey A; Sukhatme, Vikas P; Vieweg, Johannes W; Wood, Christopher G; King, Laura

    2007-01-15

    Innovations and Challenges in Renal Cancer, chaired by Michael B. Atkins, was held April 28 to 29, 2006 in Cambridge, Massachusetts. The conference brought together leading experts in the fields of cancer research, medical oncology, urology, immunology, radiology, and immunotherapy, with the goal of advancing the field of renal cancer treatment by critiquing new data from ongoing clinical trials and stimulating communication among those involved in basic and clinical research. The conference proceedings published in this educational supplement to Clinical Cancer Research are intended to provide timely information and recommendations on important aspects of renal cancer genetics and biology and advances in prognostic classification and treatment.

  1. Brief history of the Cambridge STEM aberration correction project and its progeny.

    Science.gov (United States)

    Brown, L Michael; Batson, Philip E; Dellby, Niklas; Krivanek, Ondrej L

    2015-10-01

    We provide a brief history of the project to correct the spherical aberration of the scanning transmission electron microscope (STEM) that started in Cambridge (UK) and continued in Kirkland (WA, USA), Yorktown Heights (NY, USA), and other places. We describe the project in the full context of other aberration correction research and related work, partly in response to the incomplete context presented in the paper "In quest of perfection in electron optics: A biographical sketch of Harald Rose on the occasion of his 80th birthday", recently published in Ultramicroscopy.

  2. PCR-Reverse Blot Hybridization Assay for Screening and Identification of Pathogens in Sepsis

    OpenAIRE

    Choi, Yeonim; Wang, Hye-young; Lee, Gyusang; Park, Soon-Deok; Jeon, Bo-Young; Uh, Young; Kim, Jong Bae; Lee, Hyeyoung

    2013-01-01

    Rapid and accurate identification of the pathogens involved in bloodstream infections is crucial for the prompt initiation of appropriate therapy, as this can decrease morbidity and mortality rates. A PCR-reverse blot hybridization assay for sepsis, the reverse blot hybridization assay (REBA) Sepsis-ID test, was developed; it uses pan-probes to distinguish Gram-positive and -negative bacteria and fungi. In addition, the assay was designed to identify bacteria and fungi using six genus-specifi...

  3. Accuracy of Reverse Dot-Blot PCR in Detection of Different β-Globin Gene Mutations.

    Science.gov (United States)

    El-Fadaly, N; Abd-Elhameed, A; Abd-Elbar, E; El-Shanshory, M

    2016-06-01

    Prevention programs for β-thalassemia based on molecular diagnosis of heterozygous carriers and/or patients require the use of reliable mutation screening methods. The aim of this study was to compare between direct DNA sequencing, and reverse dot-blot PCR in detection of different β-globin gene mutations in Egyptian children with β-thalassemia. Forty children with β-thalassemia were subjected to mutation analysis, performed by both direct DNA sequencing and β-globin Strip Assay MED™ (based on reverse dot-blot PCR). The most frequent mutant alleles detected by reverse dot-blot PCR were; IVSI-110 G>A (31.25 %), IVS I-6 T > C (21.25 %), and IVS I-1 G>A (20 %). Relatively less frequent mutant alleles detected by reverse dot-blot PCR were "IVSII-1 G>A (5 %), IVSII-745 C>G (5 %), IVSII-848 C>A (2.5 %), IVSI-5 G>C (2.5 %), -87 C>G(2.5 %), and cd39 C>T (2.5 %)", While the genotypes of three patients (6 alleles 7.5 %) were not detected by reverse dot-blot PCR. Mutant alleles detected by direct DNA sequencing were the same as reverse dot-blot PCR method except it revealed the genotypes of 3 undetected patients (one patient was homozygous IVSI-110 G>A, and two patients were homozygous IVS I-1 G>A. Sensitivity of the reverse dot-blot PCR was 92.5 % when compared to direct DNA sequencing for detecting β-thalassemia mutations. Our results therefore suggest that, direct DNA sequencing may be preferred over reverse dot-blot PCR in critical diagnostic situations like genetic counseling for prenatal diagnosis.

  4. Building America Case Study: Boiler Control Replacement for Hydronically Heated Multifamily Buildings, Cambridge, Massachusetts (Fact Sheet)

    Energy Technology Data Exchange (ETDEWEB)

    2014-11-01

    The ARIES Collaborative, a U.S. Department of Energy Building America research team, partnered with NeighborWorks America affiliate Homeowners' Rehab Inc. (HRI) of Cambridge, Massachusetts, to study improvements to the central hydronic heating system in one of the nonprofit's housing developments. The heating controls in the three-building, 42-unit Columbia Cambridge Alliance for Spanish Tenants housing development were upgraded. Fuel use in the development was excessive compared to similar properties. A poorly insulated thermal envelope contributed to high energy bills, but adding wall insulation was not cost-effective or practical. The more cost-effective option was improving heating system efficiency. Efficient operation of the heating system faced several obstacles, including inflexible boiler controls and failed thermostatic radiator valves. Boiler controls were replaced with systems that offer temperature setbacks and one that controls heat based on apartment temperature in addition to outdoor temperature. Utility bill analysis shows that post-retrofit weather-normalized heating energy use was reduced by 10%-31% (average of 19%). Indoor temperature cutoff reduced boiler runtime (and therefore heating fuel consumption) by 28% in the one building in which it was implemented. Nearly all savings were obtained during night which had a lower indoor temperature cut off (68 degrees F) than day (73 degrees F). This implies that the outdoor reset curve was appropriately adjusted for this building for daytime operation. Nighttime setback of heating system supply water temperature had no discernable impact on boiler runtime or gas bills.

  5. Do the Cambridge Neuropsychological Test Automated Battery episodic memory measures discriminate amnestic mild cognitive impairment?

    Science.gov (United States)

    Juncos-Rabadán, Onésimo; Pereiro, Arturo X; Facal, David; Reboredo, Alba; Lojo-Seoane, Cristina

    2014-06-01

    Although visual recognition memory and visuospatial paired associates learning has been shown to be impaired in amnestic mild cognitive impairment (aMCI), the sensitivity and specificity of the visual memory tests used to identify aMCI are not well defined. The current study attempted to analyze the sensitivity and specificity of three visual episodic memory tests (Pattern Recognition Memory [PRM], Delayed Matching to Sample [DMS], and Paired Associated Learning [PAL]) from the CANTAB, in differentiating aMCI patients from control healthy participants. Seventy seven aMCI patients and 85 cognitive normal controls aged over 50 years performed the PRM, DMS, and PAL tests. Univariate and multivariate logistic regression and receiver operating characteristic curve analyses were used to study the relationships between aMCI and visual memory measures. The three Cambridge Neuropsychological Test Automated Battery measures significantly predicted aMCI. The optimal predictive model combined the total percent correct responses for PRM and DMS with the PAL total errors (six shapes adjusted), with a sensitivity of 72%, specificity of 83%, and achieved predictive accuracy of 80%. Visual episodic memory tasks such as those involved in the PRM, DMS, and PAL tests (included in the Cambridge Neuropsychological Test Automated Battery) may sensitively discriminate aMCI patients from normal controls. These tests may be useful for correct diagnosis of aMCI. Copyright © 2013 John Wiley & Sons, Ltd.

  6. Technology Solutions Case Study: Boiler Control Replacement for Hydronically Heated Multifamily Buildings, Cambridge, Massachusetts

    Energy Technology Data Exchange (ETDEWEB)

    None

    2014-11-01

    The ARIES Collaborative, a U.S. Department of Energy Building America research team, partnered with NeighborWorks America affiliate Homeowners' Rehab Inc. (HRI) of Cambridge, Massachusetts, to study improvements to the central hydronic heating system in one of the nonprofit's housing developments. The heating controls in the three-building, 42-unit Columbia Cambridge Alliance for Spanish Tenants housing development were upgraded. Fuel use in the development was excessive compared to similar properties. A poorly insulated thermal envelope contributed to high energy bills, but adding wall insulation was not cost-effective or practical. The more cost-effective option was improving heating system efficiency, which faced several obstacles, including inflexible boiler controls and failed thermostatic radiator valves. Boiler controls were replaced with systems that offer temperature setbacks and one that controls heat based on apartment temperature in addition to outdoor temperature. Utility bill analysis shows that post-retrofit weather-normalized heating energy use was reduced by 10%-31% (average of 19%). Indoor temperature cutoff reduced boiler runtime (and therefore heating fuel consumption) by 28% in the one building in which it was implemented. Nearly all savings were obtained during night which had a lower indoor temperature cut off (68°F) than day (73° F). This implies that the outdoor reset curve was appropriately adjusted for this building for daytime operation. Nighttime setback of heating system supply water temperature had no discernable impact on boiler runtime or gas bills.

  7. The factors influencing car use in a cycle-friendly city: the case of Cambridge.

    Science.gov (United States)

    Carse, Andrew; Goodman, Anna; Mackett, Roger L; Panter, Jenna; Ogilvie, David

    2013-04-01

    Encouraging people out of their cars and into other modes of transport, which has major advantages for health, the environment and urban development, has proved difficult. Greater understanding of the influences that lead people to use the car, particularly for shorter journeys, may help to achieve this. This paper examines the predictors of car use compared with the bicycle to explore how it may be possible to persuade more people to use the bicycle instead of the car. Multivariable logistic regression was used to examine the socio-demographic, transport and health-related correlates of mode choice for work, shopping and leisure trips in Cambridge, a city with high levels of cycling by UK standards. The key findings are that commuting distance and free workplace parking were strongly associated with use of the car for work trips, and car availability and lower levels of education were associated with car use for leisure, shopping and short-distanced commuting trips. The case of Cambridge shows that more policies could be adopted, particularly a reduction in free car parking, to increase cycling and reduce the use of the car, especially over short distances.

  8. The afterlife of Laurence Sterne (1713-1768): Body snatching, dissection and the role of Cambridge anatomist Charles Collignon.

    Science.gov (United States)

    Dittmar, Jenna M; Mitchell, Piers D

    2016-11-01

    This paper aims to highlight the practice of body snatching from graves in the 1700s for the purpose of providing corpses for anatomical dissection, and for stocking anatomy museums. To do this, we examine the exhumation and dissection of the famous eighteenth-century novelist Laurence Sterne and explore the involvement of Charles Collignon, Professor of Anatomy at the University of Cambridge. We also show that osteological and cut-mark analysis of a skull purported to be that of Sterne, currently housed in the Duckworth Collection at Cambridge, provides the key to solving the mystery surrounding why Sterne was resurrected. © The Author(s) 2015.

  9. Applications of the Cambridge Structural Database in organic chemistry and crystal chemistry.

    Science.gov (United States)

    Allen, Frank H; Motherwell, W D Samuel

    2002-06-01

    The Cambridge Structural Database (CSD) and its associated software systems have formed the basis for more than 800 research applications in structural chemistry, crystallography and the life sciences. Relevant references, dating from the mid-1970s, and brief synopses of these papers are collected in a database, DBUse, which is freely available via the CCDC website. This database has been used to review research applications of the CSD in organic chemistry, including supramolecular applications, and in organic crystal chemistry. The review concentrates on applications that have been published since 1990 and covers a wide range of topics, including structure correlation, conformational analysis, hydrogen bonding and other intermolecular interactions, studies of crystal packing, extended structural motifs, crystal engineering and polymorphism, and crystal structure prediction. Applications of CSD information in studies of crystal structure precision, the determination of crystal structures from powder diffraction data, together with applications in chemical informatics, are also discussed.

  10. Fiftieth Anniversary of the Cambridge Structural Database and Thirty Years of Its Use in Croatia

    Directory of Open Access Journals (Sweden)

    Kojić-Prodić B.

    2015-07-01

    Full Text Available This article is dedicated to the memory of Dr. F. H. Allen and the 50th anniversary of the Cambridge Crystallographic Data Centre (CCDC; the world-renowned centre for deposition and control of crystallographic data including atomic coordinates that define the three-dimensional structures of organic molecules and metal complexes containing organic ligands. The mission exposed at the web site (http://www.ccdc.cam.ac.uk is clearly stated: “The Cambridge Crystallographic Data Centre (CCDC is dedicated to the advancement of chemistry and crystallography for the public benefit through providing high quality information, software and services.” The Cambridge Structural Database (CSD, one among the first established electronic databases, nowadays is one of the most significant crystallographic databases in the world. In the International Year of Crystallography 2014, the CSD announced in December over 750,000 deposited structures. The use of the extensive and rapidly growing database needs support of sophisticated and efficient software for checking, searching, analysing, and visualising structural data. The seminal role of the CSD in researches related to crystallography, chemistry, materials science, solid state physics and chemistry, (biotechnology, life sciences, and pharmacology is widely known. The important issues of the CCDC are the accuracy of deposited data and development of software for checking the data. Therefore, the Crystallographic Information File (CIF is introduced as the standard text file format for representing crystallographic information. Among the most important software for users is ConQuest, which enables searching all the CSD information fields, and the web implementation WebCSD software. Mercury is available for visualisation of crystal structures and crystal morphology including intra- and intermolecular interactions with graph-set notations of hydrogen bonds, and analysis of geometrical parameters. The CCDC gives even

  11. The Cambridge Structural Database: a quarter of a million crystal structures and rising.

    Science.gov (United States)

    Allen, Frank H

    2002-06-01

    The Cambridge Structural Database (CSD) now contains data for more than a quarter of a million small-molecule crystal structures. The information content of the CSD, together with methods for data acquisition, processing and validation, are summarized, with particular emphasis on the chemical information added by CSD editors. Nearly 80% of new structural data arrives electronically, mostly in CIF format, and the CCDC acts as the official crystal structure data depository for 51 major journals. The CCDC now maintains both a CIF archive (more than 73,000 CIFs dating from 1996), as well as the distributed binary CSD archive; the availability of data in both archives is discussed. A statistical survey of the CSD is also presented and projections concerning future accession rates indicate that the CSD will contain at least 500,000 crystal structures by the year 2010.

  12. Reginald Crundall Punnett: first Arthur Balfour Professor of Genetics, Cambridge, 1912.

    Science.gov (United States)

    Edwards, A W F

    2012-09-01

    R. C. Punnett, the codiscoverer of linkage with W. Bateson in 1904, had the good fortune to be invited to be the first Arthur Balfour Professor of Genetics at Cambridge University, United Kingdom, in 1912 when Bateson, for whom it had been intended, declined to leave his new appointment as first Director of the John Innes Horticultural Institute. We here celebrate the centenary of the first professorship dedicated to genetics, outlining Punnett's career and his scientific contributions, with special reference to the discovery of "partial coupling" in the sweet pea (later "linkage") and to the diagram known as Punnett's square. His seeming reluctance as coauthor with Bateson to promote the reduplication hypothesis to explain the statistical evidence for linkage is stressed, as is his relationship with his successor as Arthur Balfour Professor, R. A. Fisher. The background to the establishment of the Professorship is also described.

  13. Debate on Bruce Bimber´s Book Information and American Democracy. Cambridge University Press, 2003

    Directory of Open Access Journals (Sweden)

    Karpf, David

    2011-12-01

    Full Text Available Not availablePresentation José Manuel Robles Abstract of Information and American Democracy. Cambridge University Press, 2003 Bruce Bimber From Regimes to Ecologies: Globalizing Bruce Bimber’s Model of Information and Politics Steven Livingston Internet, new forms of power and democracy José Luís Garcia Internet: A Technological Tool and Changes in Political Power Liu Gang Information and American Democracy in the era of web 2.0 Lorenzo Mosca What Comes Next?: Bimber’s Information Revolutions and Institutional Disruptions David Karpf Online Political Information and Online Political Participation José Manuel Robles Digital Media and Political Change: A Response to Garcia, Karpf, Livingston, Liu, Mosca, and Robles Bruce Bimber

  14. Cambridge Rocketry Simulator – A Stochastic Six-Degrees-of-Freedom Rocket Flight Simulator

    Directory of Open Access Journals (Sweden)

    Willem J. Eerland

    2017-02-01

    Full Text Available The Cambridge Rocketry Simulator can be used to simulate the flight of unguided rockets for both design and operational applications. The software consists of three parts: The first part is a GUI that enables the user to design a rocket. The second part is a verified and peer-reviewed physics model that simulates the rocket flight. This includes a Monte Carlo wrapper to model the uncertainty in the rocket’s dynamics and the atmospheric conditions. The third part generates visualizations of the resulting trajectories, including nominal performance and uncertainty analysis, e.g. a splash-down region with confidence bounds. The project is available on SourceForge, and is written in Java (GUI, C++ (simulation core, and Python (visualization. While all parts can be executed from the GUI, the three components share information via XML, accommodating modifications, and re-use of individual components.

  15. Possessing History and American Innocence: James Baldwin, William F. Buckley, Jr., and the 1965 Cambridge Debate

    Directory of Open Access Journals (Sweden)

    Daniel Robert McClure

    2016-12-01

    Full Text Available The 1965 debate at Cambridge University between James Baldwin and William F. Buckley, Jr., posed the question: “Has the American Dream been achieved at the Expense of the American Negro?” Within the contours of the debate, Baldwin and Buckley wrestled with the ghosts of settler colonialism and slavery in a nation founded on freedom and equality. Framing the debate within the longue durée, this essay examines the deep cultural currents related to the American racial paradox at the height of the Civil Rights movement. Underscoring the changing language of white resistance against black civil rights, the essay argues that the Baldwin and Buckley debate anticipated the ways the U.S. would address racial inequality in the aftermath of the civil rights era and the dawn of neoliberalism in the 1970s.

  16. Cheminformatics Research at the Unilever Centre for Molecular Science Informatics Cambridge.

    Science.gov (United States)

    Fuchs, Julian E; Bender, Andreas; Glen, Robert C

    2015-09-01

    The Centre for Molecular Informatics, formerly Unilever Centre for Molecular Science Informatics (UCMSI), at the University of Cambridge is a world-leading driving force in the field of cheminformatics. Since its opening in 2000 more than 300 scientific articles have fundamentally changed the field of molecular informatics. The Centre has been a key player in promoting open chemical data and semantic access. Though mainly focussing on basic research, close collaborations with industrial partners ensured real world feedback and access to high quality molecular data. A variety of tools and standard protocols have been developed and are ubiquitous in the daily practice of cheminformatics. Here, we present a retrospective of cheminformatics research performed at the UCMSI, thereby highlighting historical and recent trends in the field as well as indicating future directions.

  17. Analysis of residue conformations in peptides in Cambridge structural database and protein-peptide structural complexes.

    Science.gov (United States)

    Raghavender, Upadhyayula Surya

    2017-03-01

    A comprehensive statistical analysis of the geometric parameters of peptide chains in a reduced dataset of protein-peptide complexes in Protein Data Bank (PDB) is presented. The angular variables describing the backbone conformations of amino acid residues in peptide chains shed insights into the conformational preferences of peptide residues interacting with protein partners. Nonparametric statistical approaches are employed to evaluate the interrelationships and associations in structural variables. Grouping of residues based on their structure into chemical classes reveals characteristic trends in parameter relationships. A comparison of canonical amino acid residues in free peptide structures in Cambridge structural database (CSD) with identical residues in PDB complexes, suggests that the information can be integrated from both the structural repositories enabling efficient and accurate modeling of biologically active peptides. © 2016 John Wiley & Sons A/S.

  18. Reginald Crundall Punnett: First Arthur Balfour Professor of Genetics, Cambridge, 1912

    Science.gov (United States)

    Edwards, A. W. F.

    2012-01-01

    R. C. Punnett, the codiscoverer of linkage with W. Bateson in 1904, had the good fortune to be invited to be the first Arthur Balfour Professor of Genetics at Cambridge University, United Kingdom, in 1912 when Bateson, for whom it had been intended, declined to leave his new appointment as first Director of the John Innes Horticultural Institute. We here celebrate the centenary of the first professorship dedicated to genetics, outlining Punnett’s career and his scientific contributions, with special reference to the discovery of “partial coupling” in the sweet pea (later “linkage”) and to the diagram known as Punnett’s square. His seeming reluctance as coauthor with Bateson to promote the reduplication hypothesis to explain the statistical evidence for linkage is stressed, as is his relationship with his successor as Arthur Balfour Professor, R. A. Fisher. The background to the establishment of the Professorship is also described. PMID:22964834

  19. Does the Cambridge Automated Neuropsychological Test Battery (CANTAB) Distinguish Between Cognitive Domains in Healthy Older Adults?

    Science.gov (United States)

    Lenehan, Megan E; Summers, Mathew J; Saunders, Nichole L; Summers, Jeffery J; Vickers, James C

    2016-04-01

    The Cambridge Neuropsychological Test Automated Battery (CANTAB) is a semiautomated computer interface for assessing cognitive function. We examined whether CANTAB tests measured specific cognitive functions, using established neuropsychological tests as a reference point. A sample of 500 healthy older (M = 60.28 years, SD = 6.75) participants in the Tasmanian Healthy Brain Project completed battery of CANTAB subtests and standard paper-based neuropsychological tests. Confirmatory factor analysis identified four factors: processing speed, verbal ability, episodic memory, and working memory. However, CANTAB tests did not consistently load onto the cognitive domain factors derived from traditional measures of the same function. These results indicate that five of the six CANTAB subtests examined did not load onto single cognitive functions. These CANTAB tests may lack the sensitivity to measure discrete cognitive functions in healthy populations or may measure other cognitive domains not included in the traditional neuropsychological battery.

  20. Experience from two decades of the Cambridge Rapid Access Neurology Clinic.

    Science.gov (United States)

    Axinte, Laura T; Fiddes, Barnaby D; Donaghy, Alastair; Whyte, Adam; Allen, Chris; Sawcer, Stephen J; Adam, Robert J; Stacpoole, Sybil R L

    2015-10-01

    We report on the evolution of the rapid access neurology clinic (established in 1995) at Addenbrooke's Hospital, Cambridge. Annualised attendance data demonstrate an ever increasing demand, with primary headache disorders now accounting for more than 40% of referrals. Secondary causes of headache (including intracranial tumours, idiopathic intracranial hypertension, carotid or vertebral artery dissection and subdural haematomas) remain infrequent. In all such cases, there were additional diagnostic clues. The number of patients referred with problems related to chronic neurological diseases has fallen considerably, reflecting the roles of specialist nurses and clinics. Imaging investigation of choice shifted from computerised tomography scan (45 to 16%) towards magnetic resonance imaging (17 to 47%). Management is increasingly on an outpatient basis, often without the need for a follow-up appointment. The experience presented here should inform further development of rapid access neurology clinics across the UK and suggests the need for acute headache services, in line with those for transient ischaemic attack and first seizure.

  1. Bericht uber den 2. Internationalen Kongress fur Angewandte Linguistik. Cambridge 8.-12. IX. 1969. [Report on the Second International Congress for Applied Linguistics, Cambridge, Dec. 8-12, 1969.

    Science.gov (United States)

    Mohr, Peter

    This paper is a summary report on the Second International Congress of Applied Linguistics held in Cambridge, England in September 1969. Because of the large number of papers delivered, only a selection of the papers delivered in any one section of the Congress are considered, and the author attempts to identify current interests and trends in…

  2. Bericht uber den 2. Internationalen Kongress fur Angewandte Linguistik. Cambridge 8.-12. IX. 1969. [Report on the Second International Congress for Applied Linguistics, Cambridge, Dec. 8-12, 1969.

    Science.gov (United States)

    Mohr, Peter

    This paper is a summary report on the Second International Congress of Applied Linguistics held in Cambridge, England in September 1969. Because of the large number of papers delivered, only a selection of the papers delivered in any one section of the Congress are considered, and the author attempts to identify current interests and trends in…

  3. HOUSE DUST MITE ALLERGEN (Derp1 AND Blot5) LEVELS IN ASTHMATICS' HOME IN HONGKONG

    Institute of Scientific and Technical Information of China (English)

    Bao-qing Sun; Adrian Wu; Albert Chan; Stanley Chik; Dorothy Wong; Nan-shan Zhong

    2004-01-01

    Objective To measure Derpl and Blot5 allergen levels in asthmatics' homes in Hongkong.Methods Seventy houses were enrolled for a mite indoor environment study. Dust samples were obtained from two sites of each patients' house: bed and floor. Derpl and Blot5 levels were quantified by a two-site monoclonal antibody-based ELISA technique.Results The levels of Derpl allergens found in bed (geometric mean (GM) 3.43 μg/g of dust; 95%CI, 1.89-4.96 μg/g)and on the floor (GM 1.12 μg/g of dust; 95%CI, 0.71-1.53 μg/g) indicated significant differences (P=0.005). However, the levels of Blot5 allergens found in bed (GM 19.00 μg/g of dust; 95%CI, 0.89-38.90 μg/g) and on the floor (GM 6.14 μg/g of dust; 95%CI, 0.40-11.90 μg/g) showed no statistically significant difference. In addition, in regards to the exposure index for Derpl and Blot5 allergens found in bed and on the floor, 17.6% in bed and 8.6% on the floor had levels of Blot5 ≥ 10 μg/g of dust, higher than those obtained for Derp1 (7.2% and 0% in bed and on the floor respectively, P< 0.05); higher percentages in bed and on the floor (25.0% and 35.7%) were observed for levels of Blot5 =0 μg/g of dust as compared with Derpl in bed and on the floor (4.3% and 14.5% respectively, P< 0.05).Conclusions Derpl and Blot5 are the major allergens found in this regional study, Blot5 is a more potent allergen in Hongkong, probably reflecting the high level of exposure to Blomia tropicalis (Bt). Bt and Dermatophagoides pteronyssinus (Dp) allergens should be included for precise diagnosis and effective immuno-therapeutic treatment of mite allergy in Hongkong.

  4. A survey on worries of pregnant women - testing the German version of the Cambridge Worry Scale

    Directory of Open Access Journals (Sweden)

    Gensichen Jochen

    2009-12-01

    Full Text Available Abstract Background Pregnancy is a transition period in a woman's life characterized by increased worries and anxiety. The Cambridge Worry Scale (CWS was developed to assess the content and extent of maternal worries in pregnancy. It has been increasingly used in studies over recent years. However, a German version has not yet been developed and validated. The aim of this study was (1 to assess the extent and content of worries in pregnancy on a sample of women in Germany using a translated and adapted version of the Cambridge Worry Scale, and (2 to evaluate the psychometric properties of the German version. Methods We conducted a cross-sectional study and enrolled 344 pregnant women in the federal state of Baden-Württemberg, Germany. Women filled out structured questionnaires that contained the CWS, the Spielberger-State-Trait-Anxiety Inventory (STAI, as well as questions on their obstetric history. Antenatal records were also analyzed. Results The CWS was well understood and easy to fill in. The major worries referred to the process of giving birth (CWS mean value 2.26 and the possibility that something might be wrong with the baby (1.99, followed by coping with the new baby (1.57, going to hospital (1.29 and the possibility of going into labour too early (1.28. The internal consistency of the scale (0.80 was satisfactory, and we found a four-factor structure, similar to previous studies. Tests of convergent validity showed that the German CWS represents a different construct compared with state and trait anxiety but has the desired overlap. Conclusions The German CWS has satisfactory psychometric properties. It represents a valuable tool for use in scientific studies and is likely to be useful also to clinicians.

  5. Activity Limitation in Glaucoma: Objective Assessment by the Cambridge Glaucoma Visual Function Test.

    Science.gov (United States)

    Skalicky, Simon E; McAlinden, Colm; Khatib, Tasneem; Anthony, Louise May; Sim, Sing Yue; Martin, Keith R; Goldberg, Ivan; McCluskey, Peter

    2016-11-01

    We design and evaluate a computer-based objective simulation of activity limitation related to glaucoma. A cross-sectional study was performed involving 70 glaucoma patients and 14 controls. Mean age was 69.0 ± 10.2 years; 49 (58.3%) were male. The Cambridge Glaucoma Visual Function Test (CGVFT) was administered to all participants. Rasch analysis and criterion, convergent, and divergent validity tests assessed the psychometric properties of the CGVFT. Regression modeling was used to determine factors predictive of CGVFT person measures. Sociodemographic information, better and worse eye visual field parameters, visual acuity, contrast sensitivity, and the Rasch-analyzed Glaucoma Activity Limitation-9 (GAL-9) and Visual Function Questionnaire Utility Index (VFQUI) questionnaire responses were recorded. From 139 pilot CGVFT items, 59 had acceptable fit to the Rasch model, with acceptable precision (person separation index, 2.13) and targeting. Cambridge Glaucoma Visual Function Test person measure (logit) scores increased between controls (-0.20 ± 0.08) and patients with mild (-0.15 ± 0.08), moderate (-0.13 ± 0.10), and severe (-0.05 ± 0.10) glaucoma (P < 0.001, ANOVA) indicating good criterion validity. Correlation coefficients of 0.455 (P < 0.001) between CGVFT and GAL-9 person measures and 0.399 (P = 0.005) between CGVFT and VFQUI person measures demonstrated convergent validity. Divergent validity was suboptimal. On multivariable analysis, lower better eye mean deviation and greater age were associated with worsening CGVFT person measures (P ≤ 0.001). The CGVFT is a computerized visual challenge test administered to a cohort of glaucoma patients. It may benefit glaucoma patients, careers, health care providers, and policy makers, providing increased awareness of activity limitation due to glaucoma.

  6. The release of wastewater contaminants in the Arctic: A case study from Cambridge Bay, Nunavut, Canada.

    Science.gov (United States)

    Chaves-Barquero, Luis G; Luong, Kim Hoang; Mundy, C J; Knapp, Charles W; Hanson, Mark L; Wong, Charles S

    2016-11-01

    The treatment of municipal wastewater in the Arctic is challenging due to a variety of financial, operational, climatic and technical issues. To better understand the efficacy of current wastewater treatment in this region and the hazard posed to receiving waters, we assessed the occurrence of nutrients and contaminants (i.e., pharmaceuticals, antibiotic resistance genes) as they moved through a lagoon-based treatment system in Cambridge Bay, Nunavut, Canada. Wastewater treatment in this community is performed by the use of a lagoon-tundra wetland system that is discharged into the marine environment and is representative of current common practices throughout the region. In 2014, samples were collected before and during lagoon discharge from two locations in the main lagoon, one location downstream from the lagoon effluent and three locations offshore. Grab samples were collected to measure nutrients (e.g., total nitrogen and phosphorus) and the presence of antibiotic resistance gene-bearing microbes, and Polar Organic Chemical Integrative Samplers (POCIS) were deployed to collect passively organic contaminants in all locations. A total of six pharmaceuticals were detected from a screen of twenty-eight analytes during the study: atenolol, carbamazepine, clarithromycin, metoprolol, sulfamethoxazole and trimethoprim. The greatest concentrations of nutrients, antibiotic resistance genes (ARGs), and pharmaceuticals were found in sampling locations within the treatment lagoon. Offshore of the release point, we observed limited to no detection of pharmaceuticals and ARGs, but no change in total nitrogen and phosphorus from pre-release. We conclude that the current concentrations of monitored pharmaceuticals do not pose a significant hazard at this time to aquatic organisms in Cambridge Bay. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Edward LIPPMAN. A History of Western Musical Aesthetics

    OpenAIRE

    Piz, Antoni

    2011-01-01

    Dans son compte rendu à propos de Esthetics of Music de Dahlhaus (Cambridge University Press, 1982), Edward Lippman regrettait le manque d’ouvrages généraux sur l’esthétique musicale dans le monde anglophone. Il est probable qu’à cette époque déjà, Lippman avait le projet d’écrire un livre qui viendrait combler cette apparente lacune dans la recherche musicologique : en effet, A History of Western Musical Aesthetics offre un traitement général de la dimension esthétique de la musique, de la G...

  8. Assessing the Impact of the Cambridge International Acceleration Program on U.S. University Determinants of Success: A Multi-Level Modeling Approach

    Science.gov (United States)

    Shaw, Stuart; Warren, Jayne; Gill, Tim

    2014-01-01

    This article focuses on the research being conducted by Cambridge International Examinations (Cambridge) to ensure that its international assessments prepare students as well as other acceleration programs for continued study in U.S. colleges and universities. The study, which builds on previous freshman GPA data modeling work using data supplied…

  9. Assessing the Impact of the Cambridge International Acceleration Program on U.S. University Determinants of Success: A Multi-Level Modeling Approach

    Science.gov (United States)

    Shaw, Stuart; Warren, Jayne; Gill, Tim

    2014-01-01

    This article focuses on the research being conducted by Cambridge International Examinations (Cambridge) to ensure that its international assessments prepare students as well as other acceleration programs for continued study in U.S. colleges and universities. The study, which builds on previous freshman GPA data modeling work using data supplied…

  10. Tartu Ülikooli teadur kaitses Cambridgeì Ülikoolis doktorikraadi / Krõõt Nõges

    Index Scriptorium Estoniae

    Nõges, Krõõt

    2006-01-01

    Tartu Ülikooli filosoofia osakonna teadur ja eetikakeskuse stipendiaat Eva Piirimäe kaitses Cambridgeì Ülikoolis doktorikraadi ideede ajaloo erialal doktoritööga "Thomas Abbt (1738-1766) and the Philosophical Genesis of German Nationalism"

  11. Tartu Ülikooli teadur kaitses Cambridgeì Ülikoolis doktorikraadi / Krõõt Nõges

    Index Scriptorium Estoniae

    Nõges, Krõõt

    2006-01-01

    Tartu Ülikooli filosoofia osakonna teadur ja eetikakeskuse stipendiaat Eva Piirimäe kaitses Cambridgeì Ülikoolis doktorikraadi ideede ajaloo erialal doktoritööga "Thomas Abbt (1738-1766) and the Philosophical Genesis of German Nationalism"

  12. M-DCPS Student Performance in International Baccalaureate and Cambridge Advanced International Certificate of Education Programs. Research Brief. Volume 1102

    Science.gov (United States)

    Blazer, Christie

    2011-01-01

    This Research Brief summarizes the performance of M-DCPS students participating in the International Baccalaureate (IB) and Cambridge Advanced International Certificate of Education (AICE) programs. Outcome data are provided for the eight M-DCPS schools offering the two programs and corresponding examinations. Participation in international…

  13. Degrees of Influence: The Politics of Honorary Degrees in the Universities of Oxford and Cambridge, 1900-2000

    Science.gov (United States)

    Heffernan, Michael; Jons, Heike

    2007-01-01

    The universities of Oxford and Cambridge had developed different attitudes towards the award of honorary degrees through the early and middle decades of the twentieth century. Recently, both have adopted a similar cautious and apolitical stance. This essay describes the role of honorary degrees in the production and reproduction of their cultural…

  14. PET-blot analysis contributes to BSE strain recognition in C57Bl/6 mice.

    Science.gov (United States)

    Lezmi, Stéphane; Bencsik, Anna; Baron, Thierry

    2006-10-01

    Identification of the strain of agent responsible for bovine spongiform encephalopathy (BSE) can be made histologically through the analysis of both distribution and intensity of brain vacuolar lesions after BSE transmission to mouse. Another useful way to distinguish the BSE agent from other prion strains is the study of the distribution of the abnormal prion protein (PrP(res)). For that purpose, paraffin-embedded tissue blot (PET-blot) method was applied on brains from C57Bl/6 mice infected with cattle BSE, experimental sheep BSE, or feline spongiform encephalopathy (FSE) from a cheetah. PrP(res) distribution was comparable, whichever of the three BSE agent sources was considered and was distinct from the PrP(res) distribution in C57Bl/6 mice inoculated with a French scrapie isolate or with a mouse-adapted scrapie strain (C506M3). These data confirm a common origin of infectious agent responsible for the British and French cattle BSE. They also indicate that PET-blot method appears as a precise complementary tool in prion strain studies because it offers easy and quick assessment of the PrP(res) mapping. Advantages and limits of the PET-blot method are discussed and compared with other established and validated methods of strain typing.

  15. Miniaturized fluorescent RNA dot blot method for rapid quantitation of gene expression

    Directory of Open Access Journals (Sweden)

    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.

  16. 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...

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

    Directory of Open Access Journals (Sweden)

    Erin Curry

    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.

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

    Science.gov (United States)

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

    2016-12-26

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

  19. 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.

  20. DIFFERENTIATION OF PSEUDOCONDYLOMA OF VULVA AND CONDYLOMA ACUMINATA BY DOT BLOT HYBRIDIZATION AND POLYMERASE CHAIN REACTION

    Institute of Scientific and Technical Information of China (English)

    刘跃华; 王家璧; 司静懿

    1996-01-01

    This study differentiated pseudocondyloma of vulva from condyloma acunainata using dot blot hybridization and polymerase chain reaction (PCR). A total of 27 cases o{ pseudocondyloma of vulva and 65 cases of condyloma acuminata were selected for the sttldy. The genital lesions were examined clinically and were biopsled. Each biopsy v-as subjected to histological examination and HPV DNA analysis by dot blot hybridization and PCR. Dot blot analysis detected HPV DNA in 19(82.6%) out of 23 cases of condyloma acuminata and 2(25%) out of 8 cases pseudocondyloma of vulvae(P<0. 05). PCR detected HPV DNA in 51(92.7%) our of 55 cases of eondyloma acuminata, compared with none in 23 cases of pseudocondylorna(P<0. 001). HPV DNA was present in the majority of condyloma acuminata specimens, HPV 6 and 11 were the predominant types. Peudocondyloma is probably not associated with HPV. PCR was the most sensitive and useful techntque for HPV DNA detection.

  1. 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.

  2. Characteristic conformation of Mosher's amide elucidated using the cambridge structural database.

    Science.gov (United States)

    Ichikawa, Akio; Ono, Hiroshi; Mikata, Yuji

    2015-07-16

    Conformations of the crystalline 3,3,3-trifluoro-2-methoxy-2-phenylpropanamide derivatives (MTPA amides) deposited in the Cambridge Structural Database (CSD) were examined statistically as Racid-enantiomers. The majority of dihedral angles (48/58, ca. 83%) of the amide carbonyl groups and the trifluoromethyl groups ranged from -30° to 0° with an average angle θ1 of -13°. The other conformational properties were also clarified: (1) one of the fluorine atoms was antiperiplanar (ap) to the amide carbonyl group, forming a staggered conformation; (2) the MTPA amides prepared from primary amines showed a Z form in amide moieties; (3) in the case of the MTPA amide prepared from a primary amine possessing secondary alkyl groups (i.e., Mosher-type MTPA amide), the dihedral angles between the methine groups and the carbonyl groups were syn and indicative of a moderate conformational flexibility; (4) the phenyl plane was inclined from the O-Cchiral bond of the methoxy moiety with an average dihedral angle θ2 of +21°; (5) the methyl group of the methoxy moiety was ap to the ipso-carbon atom of the phenyl group.

  3. Characteristic Conformation of Mosher’s Amide Elucidated Using the Cambridge Structural Database

    Directory of Open Access Journals (Sweden)

    Akio Ichikawa

    2015-07-01

    Full Text Available Conformations of the crystalline 3,3,3-trifluoro-2-methoxy-2-phenylpropanamide derivatives (MTPA amides deposited in the Cambridge Structural Database (CSD were examined statistically as Racid-enantiomers. The majority of dihedral angles (48/58, ca. 83% of the amide carbonyl groups and the trifluoromethyl groups ranged from –30° to 0° with an average angle θ1 of −13°. The other conformational properties were also clarified: (1 one of the fluorine atoms was antiperiplanar (ap to the amide carbonyl group, forming a staggered conformation; (2 the MTPA amides prepared from primary amines showed a Z form in amide moieties; (3 in the case of the MTPA amide prepared from a primary amine possessing secondary alkyl groups (i.e., Mosher-type MTPA amide, the dihedral angles between the methine groups and the carbonyl groups were syn and indicative of a moderate conformational flexibility; (4 the phenyl plane was inclined from the O–Cchiral bond of the methoxy moiety with an average dihedral angle θ2 of +21°; (5 the methyl group of the methoxy moiety was ap to the ipso-carbon atom of the phenyl group.

  4. Item response theory analyses of the Cambridge Face Memory Test (CFMT).

    Science.gov (United States)

    Cho, Sun-Joo; Wilmer, Jeremy; Herzmann, Grit; McGugin, Rankin Williams; Fiset, Daniel; Van Gulick, Ana E; Ryan, Kaitlin F; Gauthier, Isabel

    2015-06-01

    We evaluated the psychometric properties of the Cambridge Face Memory Test (CFMT; Duchaine & Nakayama, 2006). First, we assessed the dimensionality of the test with a bifactor exploratory factor analysis (EFA). This EFA analysis revealed a general factor and 3 specific factors clustered by targets of CFMT. However, the 3 specific factors appeared to be minor factors that can be ignored. Second, we fit a unidimensional item response model. This item response model showed that the CFMT items could discriminate individuals at different ability levels and covered a wide range of the ability continuum. We found the CFMT to be particularly precise for a wide range of ability levels. Third, we implemented item response theory (IRT) differential item functioning (DIF) analyses for each gender group and 2 age groups (age ≤ 20 vs. age > 21). This DIF analysis suggested little evidence of consequential differential functioning on the CFMT for these groups, supporting the use of the test to compare older to younger, or male to female, individuals. Fourth, we tested for a gender difference on the latent facial recognition ability with an explanatory item response model. We found a significant but small gender difference on the latent ability for face recognition, which was higher for women than men by 0.184, at age mean 23.2, controlling for linear and quadratic age effects. Finally, we discuss the practical considerations of the use of total scores versus IRT scale scores in applications of the CFMT.

  5. Swift observations of unidentified radio sources in the revised Third Cambridge Catalogue

    Science.gov (United States)

    Maselli, A.; Massaro, F.; Cusumano, G.; La Parola, V.; Harris, D. E.; Paggi, A.; Liuzzo, E.; Tremblay, G. R.; Baum, S. A.; O'Dea, C. P.

    2016-08-01

    We have investigated a group of unassociated radio sources included in the Third Cambridge Catalogue (3CR) to increase the multifrequency information on them and possibly obtain an identification. We have carried out an observational campaign with the Swift satellite to observe with the Ultraviolet/Optical Telescope (UVOT) and the X-Ray Telescope (XRT) the field of view of 21 bright NRAO VLA Sky Survey (NVSS) sources within the positional uncertainty region of the 3CR sources. Furthermore, we have searched in the recent AllWISE Source Catalogue for infrared sources matching the position of these NVSS sources. We have detected significant emission in the soft X-ray band for nine of the investigated NVSS sources. To all of them, and in four cases with no soft X-ray association, we have associated a Wide-field Infrared Survey Explorer infrared counterpart. Eight of these infrared candidates have not been proposed earlier in the literature. In the five remaining cases our candidate matches one among a few optical candidates suggested for the same 3CR source in previous studies. No source has been detected in the UVOT filters at the position of the NVSS objects, confirming the scenario that all of them are heavily obscured. With this in mind, a spectroscopic campaign, preferably in the infrared band, will be necessary to establish the nature of the sources that we have finally identified.

  6. Marrying content and process in clinical method teaching: enhancing the Calgary-Cambridge guides.

    Science.gov (United States)

    Kurtz, Suzanne; Silverman, Jonathan; Benson, John; Draper, Juliet

    2003-08-01

    Communication skills training is now internationally accepted as an essential component of medical education. However, learners and teachers in communication skills programs continue to experience problems integrating communication with other clinical skills, ensuring that clinical faculty support and teach communication beyond the formal communication course, extending communication training coherently into clerkship and residency, and applying communication skills in medical practice at a professional level of competence. One factor contributing to these problems is that learners confront two apparently conflicting models of the medical interview: a communication model describing the process of the interview and the "traditional medical history" describing the content of the interview. The resulting confusion exacerbates the above dilemmas and interferes with learners using communication skills training to advantage in real-life practice. The authors propose a comprehensive clinical method that explicitly integrates traditional clinical method with effective communication skills. To implement this more comprehensive approach, they have modified their own Calgary-Cambridge guides to the medical interview by developing three diagrams that visually and conceptually improve the way communication skills teaching is introduced and that place communication process skills within a comprehensive clinical method; devising a content guide for medical interviewing that is more closely aligned with the structure and process skills used in communication skills training; and incorporating patient-centered medicine into both process and content aspects of the medical interview. These enhancements help resolve ongoing difficulties associated with both teaching communication skills and applying them effectively in medical practice.

  7. Prevention of retinal detachment in Stickler syndrome: the Cambridge prophylactic cryotherapy protocol.

    Science.gov (United States)

    Fincham, Gregory S; Pasea, Laura; Carroll, Christopher; McNinch, Annie M; Poulson, Arabella V; Richards, Allan J; Scott, John D; Snead, Martin P

    2014-08-01

    The Stickler syndromes are the most common causes of inherited and childhood retinal detachment; however, no consensus exists regarding the effectiveness of prophylactic intervention. We evaluate the long-term safety and efficacy of the Cambridge prophylactic cryotherapy protocol, a standardized retinal prophylactic treatment developed to prevent retinal detachment arising from giant retinal tears in type 1 Stickler syndrome. Retrospective comparative case series. Four hundred eighty seven patients with type 1 Stickler syndrome. Time to retinal detachment was compared between patients who received bilateral prophylaxis and untreated controls, with and without individual patient matching. Patients receiving unilateral prophylaxis (after fellow eye retinal detachment) were similarly compared with an appropriate control subgroup. Individual patient matching ensured equal age and follow-up between groups and that an appropriate control (who had not suffered a retinal detachment before the age at which their individually matched treatment patient underwent prophylactic treatment) was selected. Matching was blinded to outcome events. Individual patient matching protocols purposely weighted bias against the effectiveness of treatment. All treatment side effects are reported. Time to retinal detachment and side effects occurring after prophylactic treatment. The bilateral control group (n = 194) had a 7.4-fold increased risk of retinal detachment compared to the bilateral prophylaxis group (n = 229) (hazard ratio [HR], 7.40; 95% confidence interval [CI], 4.53-12.08; PCambridge prophylactic cryotherapy protocol is safe and markedly reduces the risk of retinal detachment. Copyright © 2014 American Academy of Ophthalmology. Published by Elsevier Inc. All rights reserved.

  8. Testing the Cambridge Quality Checklists on a review of disrupted families and crime

    Science.gov (United States)

    Jolliffe, Darrick; Murray, Joseph; Farrington, David; Vannick, Claire

    2012-01-01

    Background Systematic reviews of the relationship between non-manipulated factors (e.g. low empathy) and offending are becoming more common, and it is important to consider the methodological quality of studies included in such reviews. Aims To assess aspects of the reliability and validity of the Cambridge Quality Checklists, a set of three measures for examining the methodological quality of studies included in systematic reviews of risk factors for offending. Methods All 60 studies in a systematic review of disrupted families and offending were coded on the CQC and codes compared with the effect sizes derived from the studies. Results Overall, the CQC was easy to score, and the relevant information was available in most studies. The scales had high inter-rater reliability. Only 13 studies scored high on the Checklist of Correlates, 18 scored highly on the Checklist of Risk Factors and none scored highly on the Checklist of Causal Risk Factors. Generally, studies that were of lower quality had higher effect sizes. Conclusions The CQC could be a useful method of assessing the methodological quality of studies of risk factors for offending but might benefit from additional conceptual work, changes to the wording of some scales and additional levels for scoring. Copyright © 2012 John Wiley & Sons, Ltd. PMID:23192977

  9. Diagnosing prosopagnosia in East Asian individuals: Norms for the Cambridge Face Memory Test-Chinese.

    Science.gov (United States)

    McKone, Elinor; Wan, Lulu; Robbins, Rachel; Crookes, Kate; Liu, Jia

    2017-09-14

    The Cambridge Face Memory Test (CFMT) is widely accepted as providing a valid and reliable tool in diagnosing prosopagnosia (inability to recognize people's faces). Previously, large-sample norms have been available only for Caucasian-face versions, suitable for diagnosis in Caucasian observers. These are invalid for observers of different races due to potentially severe other-race effects. Here, we provide large-sample norms (N = 306) for East Asian observers on an Asian-face version (CFMT-Chinese). We also demonstrate methodological suitability of the CFMT-Chinese for prosopagnosia diagnosis (high internal reliability, approximately normal distribution, norm-score range sufficiently far above chance). Additional findings were a female advantage on mean performance, plus a difference between participants living in the East (China) or the West (international students, second-generation children of immigrants), which we suggest might reflect personality differences associated with willingness to emigrate. Finally, we demonstrate suitability of the CFMT-Chinese for individual differences studies that use correlations within the normal range.

  10. Normal range of cambridge low contrast test; a population based study.

    Science.gov (United States)

    Ostadimoghaddam, Hadi; Fotouhi, Akbar; Hashemi, Hassan; Yekta, Abbas Ali; Heravian, Javad; Abdolahinia, Tahereh; Norouzi Rad, Reza; Asgari, Soheila; Khabazkhoob, Mehdi

    2014-01-01

    To determine the range of contrast sensitivity (CS) and its determinants in a normal population, Mashhad, Iran. In this cross-sectional population based study, 4,453 individuals were invited of whom 3,132 persons agreed to participate (response rate, 70.4%). CS data from 2,449 eligible individuals were analyzed. CS was determined using the Cambridge low contrast square-wave grating test, and its associations with age, gender, best-corrected visual acuity (BCVA) and manifest refraction spherical equivalent (MRSE) refractive error, were analyzed. Mean age of the participants was 29.1±17.3 (range, 4-89) years and 66.4% were female. Mean CS was 239.6±233.3 and 234.6±228.6 cps in right and left eyes, respectively. Mean binocular CS was 310.9±249.0 cps. Multiple linear regression showed that CS was inversely correlated with older age (β=-1.1, PCambridge low-contrast grating test reported herein may serve as a reference for the general population in Iran. Our findings can be used for both research and clinical applications, particularly for evaluations of the outcomes of refractive surgery. In the current study, CS was lower in older subjects, myopic individuals and patients with lower BCVA.

  11. Face recognition performance of individuals with Asperger syndrome on the Cambridge Face Memory Test.

    Science.gov (United States)

    Hedley, Darren; Brewer, Neil; Young, Robyn

    2011-12-01

    Although face recognition deficits in individuals with Autism Spectrum Disorder (ASD), including Asperger syndrome (AS), are widely acknowledged, the empirical evidence is mixed. This in part reflects the failure to use standardized and psychometrically sound tests. We contrasted standardized face recognition scores on the Cambridge Face Memory Test (CFMT) for 34 individuals with AS with those for 42, IQ-matched non-ASD individuals, and age-standardized scores from a large Australian cohort. We also examined the influence of IQ, autistic traits, and negative affect on face recognition performance. Overall, participants with AS performed significantly worse on the CFMT than the non-ASD participants and when evaluated against standardized test norms. However, while 24% of participants with AS presented with severe face recognition impairment (>2 SDs below the mean), many individuals performed at or above the typical level for their age: 53% scored within +/- 1 SD of the mean and 9% demonstrated superior performance (>1 SD above the mean). Regression analysis provided no evidence that IQ, autistic traits, or negative affect significantly influenced face recognition: diagnostic group membership was the only significant predictor of face recognition performance. In sum, face recognition performance in ASD is on a continuum, but with average levels significantly below non-ASD levels of performance.

  12. A Workshop on Environmental Technology Assessment Held at the University of Cambridge, England Held on 24-26 April, 1985,

    Science.gov (United States)

    1985-01-01

    for Air Pollutants J J Stukel Economic Issues in the Control of Air Pollution I M Torrens Theme 2 Water Treatment Megatrends in Water Treatment...TECHNOLOGY ASSESSMENTS -c -e ne>±- a- University of Cambridge, England April 24-26, 1985 MEGATRENDS IN NATER TREATMENT TECHNOLOGIES F’. Fiessinaer, J... tourism caused load varia- •-s and agro-industry discharges could and should be controled cnemicals addition within the mechanical biological process

  13. The Cambridge Face Tracker: Accurate, Low Cost Measurement of Head Posture Using Computer Vision and Face Recognition Software

    Science.gov (United States)

    Thomas, Peter B. M.; Baltrušaitis, Tadas; Robinson, Peter; Vivian, Anthony J.

    2016-01-01

    Purpose We validate a video-based method of head posture measurement. Methods The Cambridge Face Tracker uses neural networks (constrained local neural fields) to recognize facial features in video. The relative position of these facial features is used to calculate head posture. First, we assess the accuracy of this approach against videos in three research databases where each frame is tagged with a precisely measured head posture. Second, we compare our method to a commercially available mechanical device, the Cervical Range of Motion device: four subjects each adopted 43 distinct head postures that were measured using both methods. Results The Cambridge Face Tracker achieved confident facial recognition in 92% of the approximately 38,000 frames of video from the three databases. The respective mean error in absolute head posture was 3.34°, 3.86°, and 2.81°, with a median error of 1.97°, 2.16°, and 1.96°. The accuracy decreased with more extreme head posture. Comparing The Cambridge Face Tracker to the Cervical Range of Motion Device gave correlation coefficients of 0.99 (P < 0.0001), 0.96 (P < 0.0001), and 0.99 (P < 0.0001) for yaw, pitch, and roll, respectively. Conclusions The Cambridge Face Tracker performs well under real-world conditions and within the range of normally-encountered head posture. It allows useful quantification of head posture in real time or from precaptured video. Its performance is similar to that of a clinically validated mechanical device. It has significant advantages over other approaches in that subjects do not need to wear any apparatus, and it requires only low cost, easy-to-setup consumer electronics. Translational Relevance Noncontact assessment of head posture allows more complete clinical assessment of patients, and could benefit surgical planning in future. PMID:27730008

  14. The Cambridge Face Tracker: Accurate, Low Cost Measurement of Head Posture Using Computer Vision and Face Recognition Software.

    Science.gov (United States)

    Thomas, Peter B M; Baltrušaitis, Tadas; Robinson, Peter; Vivian, Anthony J

    2016-09-01

    We validate a video-based method of head posture measurement. The Cambridge Face Tracker uses neural networks (constrained local neural fields) to recognize facial features in video. The relative position of these facial features is used to calculate head posture. First, we assess the accuracy of this approach against videos in three research databases where each frame is tagged with a precisely measured head posture. Second, we compare our method to a commercially available mechanical device, the Cervical Range of Motion device: four subjects each adopted 43 distinct head postures that were measured using both methods. The Cambridge Face Tracker achieved confident facial recognition in 92% of the approximately 38,000 frames of video from the three databases. The respective mean error in absolute head posture was 3.34°, 3.86°, and 2.81°, with a median error of 1.97°, 2.16°, and 1.96°. The accuracy decreased with more extreme head posture. Comparing The Cambridge Face Tracker to the Cervical Range of Motion Device gave correlation coefficients of 0.99 (P Cambridge Face Tracker performs well under real-world conditions and within the range of normally-encountered head posture. It allows useful quantification of head posture in real time or from precaptured video. Its performance is similar to that of a clinically validated mechanical device. It has significant advantages over other approaches in that subjects do not need to wear any apparatus, and it requires only low cost, easy-to-setup consumer electronics. Noncontact assessment of head posture allows more complete clinical assessment of patients, and could benefit surgical planning in future.

  15. A robust method of measuring other-race and other-ethnicity effects: the Cambridge Face Memory Test format.

    Directory of Open Access Journals (Sweden)

    Elinor McKone

    Full Text Available Other-race and other-ethnicity effects on face memory have remained a topic of consistent research interest over several decades, across fields including face perception, social psychology, and forensic psychology (eyewitness testimony. Here we demonstrate that the Cambridge Face Memory Test format provides a robust method for measuring these effects. Testing the Cambridge Face Memory Test original version (CFMT-original; European-ancestry faces from Boston USA and a new Cambridge Face Memory Test Chinese (CFMT-Chinese, with European and Asian observers, we report a race-of-face by race-of-observer interaction that was highly significant despite modest sample size and despite observers who had quite high exposure to the other race. We attribute this to high statistical power arising from the very high internal reliability of the tasks. This power also allows us to demonstrate a much smaller within-race other ethnicity effect, based on differences in European physiognomy between Boston faces/observers and Australian faces/observers (using the CFMT-Australian.

  16. John Howard Marsden (1803–1891 First Disney Professor of Archaeology at the University of Cambridge 1851–1865

    Directory of Open Access Journals (Sweden)

    Michael Leach

    2007-05-01

    Full Text Available Although there were ten chairs of archaeology at universities in Germany, and one in France, by the mid-nineteenth century, in Great Britain it was the amateur societies and museums (the British Museum in particular that encouraged the study of this subject. In 1851 John Disney established the first university chair in Great Britain at Cambridge University. His proposal was initially received with considerable caution by the governing body of the university, and was only accepted by the narrowest margin of eight votes to seven. His agreement with the University of Cambridge stipulated that six lectures a year should be given on the subject of ‘Classical, Medieval, and other Antiquities, the Fine Arts and all matters and things connected therewith’ (Clark 1904, 222–225. However university archaeology was slow to establish its academic credibility nationally, and it was more than thirty years before Oxford University established its chair of classical archaeology. The Cambridge Board of Anthropological Studies, which included instruction in prehistoric archaeology, was not created until 1915, and as late as 1945 there were still only a few university lecturers in archaeology in Great Britain. It was not until 1946 that Oxford University appointed a Professor of Prehistoric Archaeology (Wilson 2002, 153; Daniel 1976, 6–12; Smith 2004, 4–5, 53–54.

  17. A robust method of measuring other-race and other-ethnicity effects: the Cambridge Face Memory Test format.

    Science.gov (United States)

    McKone, Elinor; Stokes, Sacha; Liu, Jia; Cohan, Sarah; Fiorentini, Chiara; Pidcock, Madeleine; Yovel, Galit; Broughton, Mary; Pelleg, Michel

    2012-01-01

    Other-race and other-ethnicity effects on face memory have remained a topic of consistent research interest over several decades, across fields including face perception, social psychology, and forensic psychology (eyewitness testimony). Here we demonstrate that the Cambridge Face Memory Test format provides a robust method for measuring these effects. Testing the Cambridge Face Memory Test original version (CFMT-original; European-ancestry faces from Boston USA) and a new Cambridge Face Memory Test Chinese (CFMT-Chinese), with European and Asian observers, we report a race-of-face by race-of-observer interaction that was highly significant despite modest sample size and despite observers who had quite high exposure to the other race. We attribute this to high statistical power arising from the very high internal reliability of the tasks. This power also allows us to demonstrate a much smaller within-race other ethnicity effect, based on differences in European physiognomy between Boston faces/observers and Australian faces/observers (using the CFMT-Australian).

  18. 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.

  19. Autoantibody profiling of patients with primary biliary cirrhosis using a multiplexed line-blot assay.

    Science.gov (United States)

    Villalta, Danilo; Sorrentino, Maria Concetta; Girolami, Elia; Tampoia, Marilina; Alessio, Maria Grazia; Brusca, Ignazio; Daves, Massimo; Porcelli, Brunetta; Barberio, Giuseppina; Bizzaro, Nicola

    2015-01-01

    To evaluate the autoantibody profile in patients with primary biliary cirrhosis (PBC) using a new multiplexed line-blot assay specifically designed for the diagnosis of autoimmune liver diseases. Sera of 58 consecutive PBC patients and 191 disease controls (144 with autoimmune liver diseases other than PBC, and 67 with non-autoimmune chronic liver diseases) were tested by both the multiplexed line-blot Autoimmune Liver Disease Profile 2 (ALD2) and by IIF on HEp-2 cells and on rat kidney/liver/stomach tissues. ALD2 contains the following PBC-associated antigens: AMA-M2, natively purified from bovine heart; M2-E3, a recombinant fusion protein including the E2 subunits of PDC, BCOADC and OGDC; sp100, PML and gp210 recombinant proteins. With the ALD2 assay, a positive reaction to AMA-M2, M2-E3, sp100, PML and gp210 in PBC patients was observed in 77.6%, 84.5%, 34.5%, 15.1% and 18.9%, respectively, of the PBC sera. The overall sensitivity and specificity for PBC were 98.3% and 93.7%. Using IIF, positivity rates to AMA, and to antinuclear autoantibodies with membranous/rim-like and multiple nuclear dot patterns were 86.2%, 8.6% and 22.4%, respectively. The overall sensitivity and specificity for PBC of the IIF method were 86.2% and 97.9%, respectively. The ALD2 line-blot showed a good diagnostic accuracy for PBC and a higher sensitivity than the IIF method to detect sp100 and gp210 autoantibodies. Copyright © 2014 Elsevier B.V. All rights reserved.

  20. Binding of 16S rRNA to chloroplast 30S ribosomal proteins blotted on nitrocellulose

    OpenAIRE

    Rozier, Claude; Mache, Régis

    1984-01-01

    Protein-RNA associations were studied by a method using proteins blotted on a nitrocellulose sheet. This method was assayed with Escherichia Coli 30S ribosomal components. In stringent conditions (300 mM NaCl or 20° C) only 9 E. coli ribosomal proteins strongly bound to the 16S rRNA: S4, S5, S7, S9, S12, S13, S14, S19, S20. 8 of these proteins have been previously found to bind independently to the 16S rRNA. The same method was applied to determine protein-RNA interactions in spinach chloropl...

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

    DEFF Research Database (Denmark)

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

    2017-01-01

    The generation of genetically engineered mouse models (GEMMs), including knock-out (KO) and knock-in (KI) models, often requires genomic screening of many mouse ES cell (mESC) clones by Southern blot. The use of large targeting constructs facilitates the recombination of exogenous DNA in a specific...... that match exons flanking the targeting construct. The detection of the expected modification in this PCR product confirms the integration at the correct genomic location and shows that the mutant mRNA is expressed. This is a simple and sensitive strategy to screen locus-specific recombination of targeting...

  2. Arvustus. Katri Lõhmus. Caring Autonomy. European Human Rights Law and the Challenge of Individualism. Cambridge University Press 2015, 246 lk / Lauri Mälksoo

    Index Scriptorium Estoniae

    Mälksoo, Lauri, 1975-

    2016-01-01

    Arvustus: Katri Lõhmus. Caring Autonomy. European Human Rights Law and the Challenge of Individualism. Cambridge University Press 2015, 246 lk. Euroopa inimõiguste ja põhivabaduste kaitse konventsiooni artikli 8 sisustamisest

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

    NARCIS (Netherlands)

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

    1997-01-01

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

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

    NARCIS (Netherlands)

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

    1997-01-01

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

  5. The German adaptation of the Cambridge pulmonary hypertension outcome review (CAMPHOR

    Directory of Open Access Journals (Sweden)

    Cima Katharina

    2012-09-01

    Full Text Available Abstract Background Individuals with precapillary pulmonary hypertension (PH experience severely impaired quality of life. A disease-specific outcome measure for PH, the Cambridge Pulmonary Hypertension Outcome Review (CAMPHOR was developed and validated in the UK and subsequently adapted for use in additional countries. The aim of this study was to translate and assess the reliability and validity of the CAMPHOR for German-speaking populations. Methods Three main adaptation stages involved; translation (employing bilingual and lay panels, cognitive debriefing interviews with patients and validation (assessment of the adaptation’s psychometric properties. The psychometric evaluation included 107 patients with precapillary PH (60 females; age mean (standard deviation 60 (15 years from 3 centres in Austria, Germany and Switzerland. Results No major problems were found with the translation process with most items easily rendered into acceptable German. Participants in the cognitive debriefing interviews found the questionnaires relevant, comprehensive and easy to complete. Psychometric analyses showed that the adaptation was successful. The three CAMPHOR scales (symptoms, activity limitations and quality of life had excellent test-retest reliability correlations (Symptoms = 0.91; Activity limitations = 0.91; QoL = 0.90 and internal consistency (Symptoms = 0.94; Activity limitations = 0.93; QoL = 0.94. Predicted correlations with the Nottingham Health Profile provided evidence of the construct validity of the CAMPHOR scales. The CAMPHOR adaptation also showed known group validity in its ability to distinguish between participants based on perceived general health, perceived disease severity, oxygen use and NYHA classification. Conclusions The CAMPHOR has been shown to be valid and reliable in the German population and is recommend for use in clinical practice.

  6. Evaluation of a brief anti-stigma campaign in Cambridge: do short-term campaigns work?

    Directory of Open Access Journals (Sweden)

    Henderson Claire

    2010-06-01

    Full Text Available Abstract Background In view of the high costs of mass-media campaigns, it is important to understand whether it is possible for a media campaign to have significant population effects over a short period of time. This paper explores this question specifically in reference to stigma and discrimination against people with mental health problems using the Time to Change Cambridge anti-stigma campaign as an example. Methods 410 face-to-face interviews were performed pre, during and post campaign activity to assess campaign awareness and mental health-related knowledge, attitudes and behaviours. Results Although campaign awareness was not sustained following campaign activity, significant and sustained shifts occurred for mental health-related knowledge items. Specifically, there was a 24% (p If a friend had a mental health problem, I know what advice to give them to get professional help, following the campaign. Additionally, for the statement: Medication can be an effective treatment for people with mental health problems, there was a 10% rise (p = 0.05 in the proportion of interviewees responding 'agree' or 'strongly agree' following the campaign. These changes, however, were not evident for attitudinal or behaviour related questions. Conclusions Although these results only reflect the impact of one small scale campaign, these preliminary findings suggest several considerations for mass-media campaign development and evaluation strategies such as: (1 Aiming to influence outcomes pertaining to knowledge in the short term; (2 Planning realistic and targeted outcomes over the short, medium and long term during sustained campaigns; and (3 Monitoring indirect campaign effects such as social discourse or other social networking/contact in the evaluation.

  7. Adaptation and Validation of the Cambridge Pulmonary Hypertension Outcome Review (CAMPHOR) for Use in Spain.

    Science.gov (United States)

    Aguirre-Camacho, Aldo; Stepanous, Jessica; Blanco-Donoso, Luis M; Moreno-Jiménez, Bernardo; Wilburn, Jeanette; González-Saiz, Laura; McKenna, Stephen P

    2017-06-01

    The Cambridge Pulmonary Hypertension Outcome Review (CAMPHOR) is a patient-reported outcome measure of health-related quality of life and quality of life specific to individuals with pulmonary hypertension (PH). This questionnaire has demonstrated superiority over other instruments assessing similar domains. The objective of the present study was to adapt and validate the Spanish version of the questionnaire. The adaptation consisted of 3 stages: translation from English to Spanish using bilingual and lay panels, cognitive debriefing interviews with patients, and assessment of psychometric properties by means of a postal validation survey. The translation panels produced a version of the CAMPHOR that was considered suitable for use by Spanish PH patients. The relevance, comprehensiveness, and acceptability of this version were confirmed in interviews with PH patients. Finally, the validation survey (n = 70) revealed that the 3 CAMPHOR scales (Symptoms, Activities, and Quality of life) showed strong psychometric properties. The internal consistency (Cronbach α) coefficients of the scales were above 0.89, and the test-retest reliability was above 0.87. The convergent and known group validity of the CAMPHOR scales was also demonstrated. The Spanish version of the CAMPHOR is a valid and reliable instrument for the assessment of health-related quality of life and quality of life in Spanish PH patients. Therefore, it is recommended for use in future research and clinical practice in the Spanish population of PH patients. Copyright © 2016 Sociedad Española de Cardiología. Published by Elsevier España, S.L.U. All rights reserved.

  8. The Cambridge Breast Intensity-modulated Radiotherapy Trial: Comparison of Clinician- versus Patient-reported Outcomes.

    Science.gov (United States)

    Mukesh, M B; Qian, W; Wah Hak, C C; Wilkinson, J S; Barnett, G C; Moody, A M; Wilson, C; Coles, C E

    2016-06-01

    Breast radiotherapy-associated toxicity is often reported using clinical and photographic assessments. The addition of patient-reported outcome measures (PROMs) is becoming more common. This study investigated the concordance between clinician- and patient-reported outcomes. The Cambridge Breast Intensity-modulated Radiotherapy (IMRT) trial prospectively collected data on clinician assessment and PROMs at 2 and 5 years after breast radiotherapy. Clinician assessment included physical examination and photographic assessment. PROMs included European Organization for Research and Treatment of Cancer (EORTC) BR23 questionnaire and four breast radiotherapy-specific questions. The correlation between patient and clinician scores were analysed on an independent patient basis using percentage agreement, Cohen's kappa coefficient (k) and Bowker's test of symmetry. The analysis was repeated after stratifying patients based on age, baseline Hospital Anxiety and Depression Score (HADS) and baseline body image score. At 2 and 5 years, a weak level of concordance was seen between the clinician-based assessment and PROMS for all the five toxicity end points (k = 0.05-0.21), with individual patient-based agreement of 32.9-78.3% and a highly discordant Bowker's test of symmetry (P < 0.001). The most frequently reported moderate-severe toxicity by patients was change in breast appearance (14% at both 2 and 5 years), whereas it was breast induration (36% and 25% at 2 and 5 years, respectively) by the clinicians. The lack of concordance was not affected by patient's age, baseline HADS and baseline body image score. This study found that moderate-severe toxicity reported by patients is low and the overall concordance between clinicians and patients is low. This could be due to methodological limitations or alternatively reflects the subjective nature of PROMs. Incorporation of a patient's perception on treatment-related toxicity will have important implications for treatment

  9. Validation of the Cambridge Prospective Memory Test (Hong Kong Chinese version) for people with stroke.

    Science.gov (United States)

    Man, David W K; Chan, M K L; Yip, C C K

    2015-01-01

    This study aimed to develop and evaluate a Hong Kong Chinese version of the Cambridge Prospective Memory Test (CAMPROMPT-HKCV). Thirty-three subjects at least one year post-stroke participated in the study. They were simultaneously rated on version A of the CAMPROMPT-HKCV by two testers to establish its internal consistency and inter-rater reliability. Raters used the parallel versions of the test (A and B), in rating 10 patients within 2 weeks to establish the parallel form reliability. Another 10 were also assessed on the same day using both version A of the CAMPROMPT-HKCV and the Rivermead Behavioural Memory Test-Chinese version (RBMT-CV) to establish concurrent validity. A new group of 40 stroke patients and 44 healthy controls was recruited to establish its sensitivity and specificity. Results indicated that test-retest reliability on time-based, event-based and total scores, and inter-rater reliability for versions A and B of the test were high. Cronbach's alpha of the event-based score was higher than that of the time-based score. The reliability and concurrent validity of the parallel forms were established. There was a significant difference in performance on CAMPROMPT-HKCV (version A) between the stroke group and the healthy control group. ROC analysis showed that the ability of the cut-off CAMPROMPT-HKCV (total score) to differentiate PM problems was 20.5 (out of 36) with sensitivity at 95.5% and specificity at 55.9%. Further study in developing stratified norms across different age groups in Chinese-speaking stroke patients is recommended.

  10. Bone remodeling around the Cambridge cup: a DEXA study of 50 hips over 2 years.

    Science.gov (United States)

    Field, Richard E; Cronin, Michael D; Singh, Parminder J; Burtenshaw, Christine; Rushton, Neil

    2006-10-01

    In a prospective 2-year study we have used dual-energy X-ray absorptiometry to measure periprosthetic bone mineral density (BMD) following implantation of a novel, "physiological", acetabular component designed using composite materials. The acetabular components were implanted in hydroxyapatite (HA) and HA-removed options. They were implanted in conjunction with a cemented femoral component in 50 female patients who presented with displaced, subcapital, fractures of the neck of the femur. Regions of interest (ROI) were defined according to De Lee and Charnley. BMD during follow-up was compared with immediate postoperative values for the affected limb. The mean precision error (CV%) was 1.01%, 2.26% and 1.12%, for ROI I, II and III respectively. The mean change in BMD, for both cups, was analyzed. There was no significant difference between the BMD changes induced with the HA- and non-HA-coated cups. After an initial fall in BMD in all 3 ROI at 6 months, ROI I and ROI II showed return to baseline BMD by 2 years. ROI III showed no significant decrease in BMD beyond 6 months, but did not return to baseline levels. Statistical analysis revealed no significant decrease in BMD in ROI I and ROI II at 2 years, compared with immediate postoperative values. The changes in BMD reflect a pattern of maximally reduced stress in the non-weight-bearing zone (ROI III), with preservation of bone density in weight bearing zones ROI I and ROI II. These results support the design principles of the Cambridge cup.

  11. Hindi translation and validation of Cambridge-Hopkins Diagnostic Questionnaire for RLS (CHRLSq

    Directory of Open Access Journals (Sweden)

    Ravi Gupta

    2015-01-01

    Full Text Available Background: Restless legs syndrome also known as Willis-Ekbom′s Disease (RLS/WED is a common illness. Cambridge-Hopkins diagnostic questionnaire for RLS (CHRLSq is a good diagnostic tool and can be used in the epidemiological studies. However, its Hindi version is not available. Thus, this study was conducted to translate and validate it in the Hindi speaking population. Materials and Methods: After obtaining the permission from the author of the CHRLSq, it was translated into Hindi language by two independent translators. After a series of forward and back translations, the finalized Hindi version was administered to two groups by one of the authors, who were blinded to the clinical diagnosis. First group consisted of RLS/WED patients, where diagnosis was made upon face to face interview and the other group - the control group included subjects with somatic symptoms disorders or exertional myalgia or chronic insomnia. Each group had 30 subjects. Diagnosis made on CHRLSq was compared with the clinical diagnosis. Statistical Analysis: Analysis was done using Statistical Package for Social Sciences (SPSS v 21.0. Descriptive statistics was calculated. Proportions were compared using chi-square test; whereas, categorical variables were compared using independent sample t-test. Sensitivity, specificity, and positive predictive value of the translated version of questionnaire were calculated. Results: Average age was comparable between the cases and control group (RLS/WED = 39.1 ± 10.1 years vs 36.2 ± 11.4 years in controls; P = 0.29. Women outnumbered men in the RLS/WED group (87% in RLS/WED group vs 57% among controls; χ2 = 6.64; P = 0.01. Both the sensitivity and specificity of the translated version was 83.3%. It had the positive predictive value of 86.6%. Conclusion: Hindi version of CHRLSq has positive predictive value of 87% and it can be used to diagnose RLS in Hindi speaking population.

  12. Cambridge-Cranfield High Performance Computing Facility (HPCF) purchases ten Sun Fire(TM) 15K servers to dramatically increase power of eScience research

    CERN Multimedia

    2002-01-01

    "The Cambridge-Cranfield High Performance Computing Facility (HPCF), a collaborative environment for data and numerical intensive computing privately run by the University of Cambridge and Cranfield University, has purchased 10 Sun Fire(TM) 15K servers from Sun Microsystems, Inc.. The total investment, which includes more than $40 million in Sun technology, will dramatically increase the computing power, reliability, availability and scalability of the HPCF" (1 page).

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

    Science.gov (United States)

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

    2010-04-01

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

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

    Science.gov (United States)

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

    2017-01-19

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

  15. Simultaneous preparation of RNA and nuclei for Northern blot and flow cytometric analysis

    Energy Technology Data Exchange (ETDEWEB)

    Tesfaigzi, J.; Jaramillo, R. [Inhalation Toxicology Research Inst., Albuquerque, NM (United States)

    1996-11-01

    Several methods have been developed to quantify RNA synthesis during the progression of the cell cycle. The rate of RNA synthesis can be detected during different stages of the cell cycle by staining cells with agents that intercalate with nucleic acids. For example, following staining of mammalian cells with acridin orange, the green and red fluorescence that correlates with DNA and RNA content, respectively, can be analyzed by flow cytometry. Increase in RNA content during the progression of cells through the cell cycle can be measured after staining with acridin orange. RNA synthesis resulting from the stimulation of quiescent cells with various growth factors has also been demonstrated by labeling cells with bromo-uridine and using the anti-bromo-deoxyuridine antibody. These methods allow measurement of the overall RNA content in cells; however, they do not allow the measurement of the levels of specific mRNAs throughout the cell cycle. Current methods to quantify specific mRNAs generally require the preparation of a large number of cells (5--10 {times} 10{sup 6} cells) to carry out flow cytometric analyses and to isolate RNA for Northern blot analysis or solution hybridization. In this report, the authors describe a method of simultaneously preparing RNA and nuclei for Northern blot and flow cytometric analyses, respectively. The minimum number of nuclei required to obtain flow cytometric data and the effect of conserving nuclei in methanol for several days are also presented.

  16. Monoclonal Antibodies against Small Molecule Natural Products and Their Applications, Eastern Blotting and Knockout Extract

    Directory of Open Access Journals (Sweden)

    Yukihiro Shoyama

    2011-06-01

    Full Text Available To determine the hapten number in hapten-carrier protein conjugate matrix-assisted laser desorption/ionization (MALDI tof mass spectrometry was applied. Highly specific anti-ginsenoside Rb1 and Rg1 monoclonal antibodies (MAbs were prepared. Ginsenosides were developed on thin layer chromatography (TLC plates which were covered by a polyvinylidene difluoride (PVDF membrane resulting in blotting. The membrane was treated with NaIO4 solution to release the aldehyde group on the sugar moiety of the ginsenosides. By treatment of the membrane with a protein solution the ginsenoside-protein conjugation as a Schiff-base occurred, which can function to fix it to the PVDF membrane. A part of the ginsenoside aglycone was reacted with anti-ginsenoside Rb1 MAb, secondary MAb conjugated with enzyme and finally a substrate was added, resulting in a specific and highly sensitive staining that we named Eastern blotting. Furthermore, it makes one-step isolation of ginsenoside Rb1 possible using an immuno-affinity column conjugated with anti-ginsenoside Rb1 MAb. Furthermore, immunoaffinity concentration was carried out allowing high sensitivity analysis of lower concentrations of ginsenoside Rb1 so that several unknown bands could be structurally determined.

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

    Directory of Open Access Journals (Sweden)

    N.S. Sato

    2004-07-01

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

  18. The Antiquarian and the Myth ofAntiquity: The Origins of Rome in Renaissance Thought, by Philip Jacks, Cambridge University Press, Cambridge

    Directory of Open Access Journals (Sweden)

    James E. Snead

    1995-11-01

    Full Text Available Attempts to understand the history of archaeology must inevitably face the issue of origins; while historical consciousness is common to human society, studying the past through the detailed excavation, analysis, and acquisition of its material remains has until recent generations been largely the domain of European society and its descendants. There is nothing inherently logical about this pursuit; the evolution of archaeology as a way of knowing the past has specifIc historical roots and antecedents in western society and needs to be understood in light of these circumstances. The Renaissance is arguably the cultural hearth in which archaeology took shape. Scholars of the age who seem to anticipate our own predilections. such as Cyriaco D'Ancona, are usually to be found in introductory chapters of archaeological textbooks immediately following Nabonidus of Babylon. Jacks takes a more contextual approach, discussing the relationships between scholarship and society in Italy from the late Medieval period through the 16th century. This is not a general history of antiquar­ian thought during the Renaissance, nor is it a study of the work of a specific individual. The author takes as his focus the changing perceptions of antiquity held by scholars of the age, with particular emphasis on the construction and manipulation of images of ancient Rome.

  19. Executive Functions as Endophenotypes in ADHD: Evidence from the Cambridge Neuropsychological Test Battery (CANTAB)

    Science.gov (United States)

    Gau, Susan Shur-Fen; Shang, Chi-Yung

    2010-01-01

    Background: Little is known about executive functions among unaffected siblings of children with attention deficit/hyperactivity disorder (ADHD), and there is lack of such information from non-Western countries. We examined verbal and nonverbal executive functions in adolescents with ADHD, unaffected siblings and controls to test whether executive…

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

    Science.gov (United States)

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

    2017-01-01

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

  1. Reliability and validity of the Cambridge Worry Scale in pregnant Turkish women.

    Science.gov (United States)

    Gunay, Emine Yigit; Gul, Asiye

    2015-03-01

    this study examined the psychometric properties of the Cambridge Worry Scale (CWS) and assessed worries in pregnant Turkish women. a descriptive, cross sectional study. the 35 Family Health Centres located in the Yıldırım sub-province of Bursa. the data were collected using the 'Pregnant Information Form', which determines the individual characteristics of the women and the 'CWS', which determines worries. The CWS is a Likert-type scale that consists of 16 items and has four sub-dimensions including the women's own health, relationships, socio-medical and socio-economic conditions. In the CWS, the total score is not calculated, and each article is evaluated in itself. 200 pregnant women were recruited from December 2010 to November 2011. The mean age of the pregnant women was 25.92 ± 5.33, 43.0% had completed primary school, and 69.0% were not in paid employment. It has been determined that the content validity index for the Turkish form is 0.98 and that the internal consistency of Cronbach's alpha value of the scale is 0.795. As a result of exploratory factor analysis, it has been concluded that the factor loadings of the scale from 0.435 to 0.902, and it can be used in a particular dimension that is not divided into the components of the scale. On the basis of the confirmatory factor analysis, it has been determined that the Goodness of Fit Index of the one-factor structure is better than four-factor structure, but the values of the goodness fit index in each model are under 0.85 and the inaccuracy of the fit index is high. the Turkish form of the CWS is an appropriate measurement tool in terms of language and content validity, and its single-factor structure can be applied to Turkish culture and can correctly identify the worries of pregnant women. Copyright © 2014 Elsevier Ltd. All rights reserved.

  2. 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.

  3. 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 10(5) 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.

  4. 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.

  5. Hydrologic, Water-Quality, and Meteorological Data for the Cambridge, Massachusetts, Drinking-Water Source Area, Water Year 2006

    Science.gov (United States)

    Smith, Kirk P.

    2008-01-01

    Records of water quantity, water quality, and meteorological parameters were continuously collected from three reservoirs, two primary streams, and four subbasin tributaries in the Cambridge, Massachusetts, drinking-water source area during water year 2006 (October 2005 through September 2006). Water samples were collected during base-flow conditions and storms in the subbasins of the Cambridge Reservoir and Stony Brook Reservoir drainage areas and analyzed for dissolved calcium, sodium, chloride, and sulfate; total nitrogen and phosphorus; and polar pesticides and metabolites. These data were collected to assist watershed administrators in managing the drinking-water source area and to identify potential sources of contaminants and trends in contaminant loading to the water supply. Monthly reservoir contents for the Cambridge Reservoir varied from about 59 to 98 percent of capacity during water year 2006, while monthly reservoir contents for the Stony Brook Reservoir and the Fresh Pond Reservoir was maintained at greater than 83 and 94 percent of capacity, respectively. If water demand is assumed to be 15 million gallons per day by the city of Cambridge, the volume of water released from the Stony Brook Reservoir to the Charles River during the 2006 water year is equivalent to an annual water surplus of about 127 percent. Recorded precipitation in the source area was about 16 percent greater for the 2006 water year than for the previous water year and was between 12 and 73 percent greater than for any recorded amount since water year 2002. The monthly mean specific-conductance values for all continuously monitored stations within the drinking-water source area were generally within the range of historical data collected since water year 1997, and in many cases were less than the historical medians. The annual mean specific conductance of 738 uS/cm (microsiemens per centimeter) for water discharged from the Cambridge Reservoir was nearly identical to the annual

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

    Directory of Open Access Journals (Sweden)

    Eduardo Miranda-Ulloa

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

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

    Directory of Open Access Journals (Sweden)

    Eduardo Miranda-Ulloa

    2014-06-01

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

  8. La Escuela de Cambridge: Historia del Pensamiento Político. Una búsqueda metodológica

    OpenAIRE

    Rabasa Gamboa, Emilio

    2016-01-01

    El artículo se centra en el análisis de la revolución metodológica que se generó el siglo pasado en la Universidad de Cambridge, Inglaterra, en la disciplina de Historia del Pensamiento Político con el fin de darle una nueva base filosófica y mayor rigor científico. Consistió en sustituir el textualismo como interpretación subjetiva de los textos clásicos, por el enfoque contextualista que explora las condiciones históricas (incluyendo el lenguaje) que determinaron el surgimiento de las ideas...

  9. Cyrtarachne keralensis Jose, 2011 is a junior synonym of Anepsion maritatum (O. Pickard-Cambridge, 1877) (Araneae, Araneidae).

    Science.gov (United States)

    Malamel, Jobi J; Sankaran, Pradeep M; Joseph, Mathew M; Sebastian, Pothalil A

    2015-11-05

    The Indo-pacific araneid genus Anepsion, with A. rhomboides (L. Koch, 1867) as the type species, was erected by Strand in 1929. He proposed the name Anepsion as a replacement name for Anepsia L. Koch, 1871, preoccupied by Anepsia Gistl, 1848, a dipteran genus (OBIS Australia, 2015). The genus was revised by Chrysanthus (1961, 1969) and currently has 16 described species and 1 subspecies (World Spider Catalog 2015). In the present paper, we are reporting the genus from India for the first time and synonymising Cyrtarachne keralensis Jose, 2011 with Anepsion maritatum O. Pickard-Cambridge, 1877. A redescription and illustrations of both male and female of A. maritatum are provided.

  10. “In my end is my beginning”. Una discussione sul caso trascurato dei Cambridge Ritualists fra antropologia comparativa, filosofia e pensiero scientifico - “In my end is my beginning”. An argument on the Cambridge Ritualists’ neglected case, on the wave of comparative anthropology, philosophy and scientific thought

    National Research Council Canada - National Science Library

    Danila Cinellu

    2015-01-01

    ... Wilson’s biophilia hypothesis and Rappaport’s engaged anthropology as springboards, this article wants to cast light on how two anthropologically undervalued manifestos of the Cambridge School – Harrison’s Themis (1912...

  11. The Cambridge Face Memory Test for Children (CFMT-C): a new tool for measuring face recognition skills in childhood.

    Science.gov (United States)

    Croydon, Abigail; Pimperton, Hannah; Ewing, Louise; Duchaine, Brad C; Pellicano, Elizabeth

    2014-09-01

    Face recognition ability follows a lengthy developmental course, not reaching maturity until well into adulthood. Valid and reliable assessments of face recognition memory ability are necessary to examine patterns of ability and disability in face processing, yet there is a dearth of such assessments for children. We modified a well-known test of face memory in adults, the Cambridge Face Memory Test (Duchaine & Nakayama, 2006, Neuropsychologia, 44, 576-585), to make it developmentally appropriate for children. To establish its utility, we administered either the upright or inverted versions of the computerised Cambridge Face Memory Test - Children (CFMT-C) to 401 children aged between 5 and 12 years. Our results show that the CFMT-C is sufficiently sensitive to demonstrate age-related gains in the recognition of unfamiliar upright and inverted faces, does not suffer from ceiling or floor effects, generates robust inversion effects, and is capable of detecting difficulties in face memory in children diagnosed with autism. Together, these findings indicate that the CFMT-C constitutes a new valid assessment tool for children's face recognition skills.

  12. Diagnosis of mild chronic pancreatitis (Cambridge classification): Comparative study using secretin injection-magnetic resonance cholangiopancreatography and endoscopic retrograde pancreatography

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    AIM: To investigate the usefulness of secretin injection MRCP for the diagnosis of mild chronic pancreatitis. METHODS: Sixteen patients having mild chronic pancreatitis according to the Cambridge classification and 12 control subjects with no abnormal findings on the pancreatogram were examined for the diagnostic accuracy of secretin injection-MRCP regarding abnormal branch pancreatic ducts associated with mild chronic pancreatitis (Cambridge Classification), using endoscopic retrograde cholangiopancreatography (ERCP) for comparison. RESULTS: The sensitivity and specificity for abnormal branch pancreatic ducts determined by two reviewers were respectively 55%-63% and 75%-83% in the head, 57%-64% and 82%-83% in the body, and 44%-44%and 72%-76% in the tail of the pancreas. The sensitivity and specificity for mild chronic pancreatitis were 56%-63% and 92%-92%, respectively. Interobserver abnormal branch pancreatic duct and of mild chronic pancreatitis was good to excellent. CONCLUSION: Secretin injection-MRCP might be useful for the diagnosis of mild chronic pancreatitis.

  13. Nano Si preparation by constant cell voltage electrolysis of FFC-Cambridge Process in molten CaCl2

    Institute of Scientific and Technical Information of China (English)

    Ji; Zhao; Shengmei; Lu; Linyan; Hu; Can; Li

    2013-01-01

    Using FFC-Cambridge Process to prepare Si from SiO2 is a promising method to prepare nanostructured and highly pure silicon for solar cells.However,the method still has many problems unsolved and the controlling effect of the cell voltage on silicon product is not clear.Here we report in this article that nano cluster-like silicon product with purity of 99.95%has been prepared by complete conversion of raw material SiO2,quartz glass plate,using constant cell voltage electrolysis FFC-Cambridge Process.By analysis of XRD,EDS,TEM,HRTEM and ICP-AES as well as the discussion from the thermodynamics calculation,the morphology and components of the product based on the change of cell voltage are clarified.It is clear that pure silicon could be prepared at the cell voltage of 1.7 2.1 V in this reaction system.The silicon material have cluster-like structure which are made of silicon nanoparticles in 20 100 nm size.Interestingly,the cluster-like nano structure of the silicon can be tuned by the used cell voltage.The purity,yield and the energy cost of silicon product prepared at the optimized cell voltage are discussed.The purity of the silicon product could be further improved,hence this method is promising for the preparation of solar grade silicon in future.

  14. Uma revisão das controvérsias sobre a Equação de Cambridge

    Directory of Open Access Journals (Sweden)

    José Luís Oreiro

    2005-08-01

    Full Text Available The objective of this article is to make a critical evaluation of the controversies about the Cambridge Equation both in the initial phase - when Pasinetti, Kaldor, Meade, Samuelson and Modigliani got involved - and in the second round that followed the publication of Steedman´s article, which contends that the Cambridge Equation still holds even in the case of a balanced government budget. In this setting we will argue that (i the Meade-Samuelson-Modigliani theorem (MSM theorem does not depend on specific hypotheses about the shape of the production function, being fully compatible with the problems of re-switching of techniques raised by the "Capital Controversy", which are only relevant for the stability of the golden-age equilibrium growth path; (ii the Pasinetti-Theorem is, however, more general than the MSM theorem since (a it holds true even after the relaxation of some simplifying hypotheses of its original version, which does not occur with the MSM theorem, and (b Kaldor´s version of the Pasinetti theorem holds true even in the case of the "euthanasia of capitalists".

  15. Intensive cisplatin/oral etoposide for epithelial ovarian cancer: the Cambridge Gynae-Oncology Centre experience: too toxic for relapse?

    Science.gov (United States)

    Gounaris, Ioannis; Iddawela, Mahesh; Parkinson, Christine; Pratt, Jennie; Hatcher, Helen; Basu, Bristi; Tan, Li Tee; Brenton, James D; Earl, Helena M

    2016-03-01

    Intensive cisplatin and oral etoposide for relapsed epithelial ovarian cancer (EOC), commonly known as the van der Burg (VDB) protocol, has been reported to improve response rates and progression-free survival. We report on all patients with relapsed EOC treated on the VDB protocol at the Cambridge Gynae-Oncology Centre. From the institutional databases, we identified all patients treated since 2001. We extracted demographic, clinical, treatment, and toxicity data and outcomes. We used Cox regression to identify predictors of survival. A total of 35 patients were treated on the VDB protocol. Toxicity was significant, with grade 3/4 fatigue, nausea and vomiting affecting 46, 46 and 29% of patients, respectively. Six patients had grade 3/4 infection and four (11%) deaths occurred on treatment. Efficacy was encouraging, with a radiological response rate of 43%, a median progression-free survival of 5.8 months and a median overall survival of 14.1 months. No significant difference in efficacy was seen between platinum-resistant and sensitive patients. We report significant activity of the VDB protocol in a routine clinical setting. However, the high rates of serious toxicity and treatment-related deaths among patients treated with palliative intent proved unacceptable. The Cambridge Gynae-Oncology Centre no longer uses this regimen in women with relapsed EOC.

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

    Science.gov (United States)

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

    1995-09-25

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

  17. Rapid Detection of Rifampin-resistant Clinical Isolates of Mycobacterium tuberculosis by Reverse Dot Blot Hybridization

    Institute of Scientific and Technical Information of China (English)

    GUO Qian; WAN Kang Lin; YU Yan; ZHU Yan Ling; ZHAO Xiu Qin; LIU Zhi Guang; ZHANG Yuan Yuan; LI Gui Lian; WEI Jian Hao; WU Yi Mou

    2015-01-01

    Objective A PCR-reverse dot blot hybridization (RDBH) assay was developed for rapid detection of rpoB gene mutations in‘hot mutation region’ of Mycobacterium tuberculosis (M. tuberculosis). Methods 12 oligonucleotide probes based on the wild-type and mutant genotype rpoB sequences of M. tuberculosis were designed to screen the most frequent wild-type and mutant genotypes for diagnosing RIF resistance. 300 M. tuberculosis clinical isolates were detected by RDBH, conventional drug-susceptibility testing (DST) and DNA sequencing to evaluate the RDBH assay. Results The sensitivity and specificity of the RDBH assay were 91.2%(165/181) and 98.3%(117/119), respectively, as compared to DST. When compared with DNA sequencing, the accuracy, positive predictive value (PPV) and negative predictive value (NPV) of the RDBH assay were 97.7%(293/300), 98.2%(164/167), and 97.0%(129/133), respectively. Furthermore, the results indicated that the most common mutations were in codons 531 (48.6%), 526 (25.4%), 516 (8.8%), and 511 (6.6%), and the combinative mutation rate was 15 (8.3%). One and two strains of insertion and deletion were found among all strains, respectively. Conclusion Our findings demonstrate that the RDBH assay is a rapid, simple and sensitive method for diagnosing RIF-resistant tuberculosis.

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

    Directory of Open Access Journals (Sweden)

    William H Roldán

    2009-05-01

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

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

    Science.gov (United States)

    Roldán, William H; Espinoza, Yrma A

    2009-05-01

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

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

    Science.gov (United States)

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

    2013-08-01

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

  1. Binding of 16S rRNA to chloroplast 30S ribosomal proteins blotted on nitrocellulose.

    Science.gov (United States)

    Rozier, C; Mache, R

    1984-10-11

    Protein-RNA associations were studied by a method using proteins blotted on a nitrocellulose sheet. This method was assayed with Escherichia Coli 30S ribosomal components. In stringent conditions (300 mM NaCl or 20 degrees C) only 9 E. coli ribosomal proteins strongly bound to the 16S rRNA: S4, S5, S7, S9, S12, S13, S14, S19, S20. 8 of these proteins have been previously found to bind independently to the 16S rRNA. The same method was applied to determine protein-RNA interactions in spinach chloroplast 30S ribosomal subunits. A set of only 7 proteins was bound to chloroplast rRNA in stringent conditions: chloroplast S6, S10, S11, S14, S15, S17 and S22. They also bound to E. coli 16S rRNA. This set includes 4 chloroplast-synthesized proteins: S6, S11, S15 and S22. The core particles obtained after treatment by LiCl of chloroplast 30S ribosomal subunit contained 3 proteins (S6, S10 and S14) which are included in the set of 7 binding proteins. This set of proteins probably play a part in the early steps of the assembly of the chloroplast 30S ribosomal subunit.

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

    Science.gov (United States)

    Muro, M D; Fernández, C; Moneo, I

    1996-01-01

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

  3. NORTHERN BLOT ANALYSIS OF nm23 GENE EXPRESSION IN HUMAN LUNG CANCER

    Institute of Scientific and Technical Information of China (English)

    LIU Lun-xu; ZHOU Qing-hua; SHI Ying-kang; QIN Yang; SUN Zhi-lin; SUN Ze-fang

    1999-01-01

    Objective: To investigate the role of nm23 gene expression in human lung cancer. Methods: Forty human lung cancer tissues and 19 non-cancer pulmonary tissues were studied for their nm23-H1 and nm23-H2 mRNA expression with non-radioactive Northern blot hybridization. The correlation of nm23 mRNA expression with clinical features of lung cancer was analyzed. Results: The mRNA expression of nm23-H2 gene in poorly differentiated squamous cell carcinoma was significantly decreased compared to that in moderate-high differentiated squamous cell carcinoma. The mRNA expression of nm23-H1 and nm23-H2 gene in small cell lung cancer was significantly decreased compared to that in squamous cell carcinoma. No significant difference in nm23 mRNA expression was observed between lung cancer with and without lymph node metastasis, nor was there significant difference between tumor stage. Conclusion: The mRNA expression of nm23 gene is correlated with the degree of differentiation of lung cancer, but there is no evidence of metastasis suppression effect by nm23 gene.

  4. Rapid detection of intestinal pathogens in fecal samples by an improved reverse dot blot method

    Institute of Scientific and Technical Information of China (English)

    Jian-Ming Xing; Su Zhang; Ying Du; Dan Bi; Li-Hui Yao

    2009-01-01

    AIM:To develop a new, rapid and accurate reverse dot blot (RDB) method for the detection of intestinal pathogens in fecal samples.METHODS:The 12 intestinal pathogens tested were Salmonella spp., Brucella spp., Escherichia coli O157:H7,Clostridium botulinum, Bacillus cereus,Clostridium perfringens, Vibrio parahaemolyticus,Shigella spp., Yersinia enterocolitica, Vibrio cholerae,Listeria monocytogenes and Staphylococcus aureus.The two universal primers were designed to amplify two variable regions of bacterial 16S and 23S rDNA genes from all of the 12 bacterial species tested. Five hundred and forty fecal samples from the diarrhea patients were detected using the improved RDB assay.RESULTS:The methods could identify the 12 intestinal pathogens specifically, and the detection limit was as low as 103 CFUs. The consistent detection rate of the improved RDB assay compared with the traditional culture method was up to 88.75%.CONCLUSION:The hybridization results indicated that the improved RDB assay developed was a reliable method for the detection of intestinal pathogen in fecal samples.

  5. Hydrologic, Water-Quality, and Meteorological Data for the Cambridge, Massachusetts, Drinking-Water Source Area, Water Year 2005

    Science.gov (United States)

    Smith, Kirk P.

    2007-01-01

    Records of water quantity, water quality, and meteorological parameters were continuously collected from three reservoirs, two primary streams, and four subbasin tributaries in the Cambridge, Massachusetts, drinking-water source area during water year 2005 (October 2004 through September 2005). Water samples were collected during base-flow conditions and storms in the subbasins of the Cambridge Reservoir and Stony Brook Reservoir drainage areas and analyzed for selected elements, organic constituents, suspended sediment, and Escherichia coli bacteria. These data were collected to assist watershed administrators in managing the drinking-water source area and to identify potential sources of contaminants and trends in contaminant loading to the water supply. Monthly reservoir capacities for the Cambridge Reservoir varied from about 59 to 98 percent during water year 2005, while monthly reservoir capacities for the Stony Brook Reservoir and the Fresh Pond Reservoir were maintained at capacities greater than 84 and 96 percent, respectively. Assuming a water demand of 15 million gallons per day by the city of Cambridge, the volume of water released from the Stony Brook Reservoir to the Charles River during the 2005 water year is equivalent to an annual water surplus of about 119 percent. Recorded precipitation in the source area for the 2005 water year was within 2 inches of the total annual precipitation for the previous 2 water years. The monthly mean specific conductances for the outflow of the Cambridge Reservoir were similar to historical monthly mean values. However, monthly mean specific conductances for Stony Brook near Route 20, in Waltham (U.S. Geological Survey station 01104460), which is the principal tributary feeding the Stony Brook Reservoir, were generally higher than the medians of the monthly mean specific conductances for the period of record. Similarly, monthly mean specific conductances for a small tributary to Stony Brook (U.S. Geological Survey

  6. Aerial Photography and Imagery, Ortho-Corrected, True Color Orthophotography for Cambridge, Hurlock, Secretary, and Vienna - 4" pixles, 2006., Published in 2006, 1:600 (1in=50ft) scale, Eastern Shore Regional GIS Cooperative.

    Data.gov (United States)

    NSGIC Regional | GIS Inventory — Aerial Photography and Imagery, Ortho-Corrected dataset current as of 2006. True Color Orthophotography for Cambridge, Hurlock, Secretary, and Vienna - 4" pixles,...

  7. Cytotoxic activity, DNA damage, cellular uptake, apoptosis and western blot analysis of ruthenium(II) polypyridyl complex against human lung decarcinoma A549 cell.

    Science.gov (United States)

    Lai, Shang-Hai; Jiang, Guang-Bin; Yao, Jun-Hua; Li, Wei; Han, Bing-Jie; Zhang, Cheng; Zeng, Chuan-Chuan; Liu, Yun-Jun

    2015-11-01

    A new ruthenium(II) polypyridyl complex [Ru(dmp)2(pddppn)](ClO4)2Ru1 was synthesized and characterized. The cytotoxic activity in vitro of the complex was evaluated by MTT method. Ru1 shows high effect on the inhibition of the cell growth against BEL-7402, HeLa, MG-63 and A549 cells with low IC50 values of 1.6±0.4, 9.0±0.8, 1.5±0.2 and 1.5±0.3 μM, respectively. The cellular uptake indicates that Ru1 can enter into the cytoplasm and accumulate in the cell nuclei. Ru1 can induce apoptosis in A549 cells and enhance the levels of reactive oxygen species (ROS) and induce the decrease of mitochondrial membrane potential. In addition, Ru1 can down-regulate the levels of Bcl-2, Bcl-x, Bak, and Bim expression and up-regulate the expression of Bag-1 and Bad. The complex induces apoptosis of A549 cells through an intrinsic ROS-mediated mitochondrial dysfunction pathway, which was accompanied by regulating the expression of caspases and Bcl-2 family proteins. Copyright © 2015 Elsevier Inc. All rights reserved.

  8. 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

  9. 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.

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

    Science.gov (United States)

    Hood-DeGrenier, Jennifer K.

    2008-01-01

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

  11. Detection of PrPSc in Formalin Fixed Paraffin Embedded Tissue by Western Blot Differentiates Classical Scrapie, Nor98 Scrapie, and BSE

    Science.gov (United States)

    Transmissible spongiform encephalopathies including bovine spongiform encephalopathy and scrapie are fatal neurodegenerative disorders associated with the presence of an infectious abnormal isoform of normal mammalian proteins called prions (PrP**Sc). Identification of PrP**Sc in the CNS is typicall...

  12. Western Blot und realtime PCR- basierte Identifizierung möglicher Hypertonie assoziierter Gene an einem definiertem Hypotoniemodell an Wistar-Kyoto Ratten

    OpenAIRE

    Furundzija, Vesna

    2010-01-01

    Essential hypertension is a disturbance of blood pressure regulation and can be conditioned by a number of factors. The causes can be traced back to a variety of environmental factors and a range of genes relevant to blood pressure. A number of genes have been identified as possible candidates, although it is probable that others exist. The goal of the present work lies both in verification but also in the discovery of new differentially expressed genes that regulate blood pressure. ...

  13. 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.

  14. Westernblot检测奶牛生殖系统朊蛋白的分布%Distribution of cellular prion protein in cow reproductive system revealed by Western blotting

    Institute of Scientific and Technical Information of China (English)

    张太翔; 田国宁; 张金玲; 田国华; 赵德明

    2008-01-01

    传染性海绵状脑病(TSEs)是一种可引起人和动物一系列神经退化性疾病的一种新型传染病,包括羊瘙痒病(Scrapie)、牛海绵状脑病(BSE)、鹿的慢性消耗性疾病(CWD)、水貂海绵状脑病(MSE)、猫的疯猫病(FSE)、人克雅氏病(CJD)。TSE疾病起始于正常的PrPc发生错误折叠,形成富含β片层的、抗蛋白酶水解的PrPSc。人们已经认识到PrPc向PrPSc转化是疾病发生和发展的关键。错误折叠的PrPSc有很强的聚合倾向,先形成淀粉样的小纤维,进一步聚合形成淀粉样斑块,最后发展为可被临床诊断的大脑海绵状退化变性病症。

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

    Science.gov (United States)

    Hood-DeGrenier, Jennifer K.

    2008-01-01

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

  16. Healthy travel and the socio-economic structure of car commuting in Cambridge, UK: a mixed-methods analysis.

    Science.gov (United States)

    Goodman, Anna; Guell, Cornelia; Panter, Jenna; Jones, Natalia R; Ogilvie, David

    2012-06-01

    Car use is associated with substantial health and environmental costs but research in deprived populations indicates that car access may also promote psychosocial well-being within car-oriented environments. This mixed-method (quantitative and qualitative) study examined this issue in a more affluent setting, investigating the socio-economic structure of car commuting in Cambridge, UK. Our analyses involved integrating self-reported questionnaire data from 1142 participants in the Commuting and Health in Cambridge study (collected in 2009) and in-depth interviews with 50 participants (collected 2009-2010). Even in Britain's leading 'cycling city', cars were a key resource in bridging the gap between individuals' desires and their circumstances. This applied both to long-term life goals such as home ownership and to shorter-term challenges such as illness. Yet car commuting was also subject to constraints, with rush hour traffic pushing drivers to start work earlier and with restrictions on, or charges for, workplace parking pushing drivers towards multimodal journeys (e.g. driving to a 'park-and-ride' site then walking). These patterns of car commuting were socio-economically structured in several ways. First, the gradient of housing costs made living near Cambridge more expensive, affecting who could 'afford' to cycle and perhaps making cycling the more salient local marker of Bourdieu's class distinction. Nevertheless, cars were generally affordable in this relatively affluent, highly-educated population, reducing the barrier which distance posed to labour-force participation. Finally, having the option of starting work early required flexible hours, a form of job control which in Britain is more common among higher occupational classes. Following a social model of disability, we conclude that socio-economic advantage can make car-oriented environments less disabling via both greater affluence and greater job control, and in ways manifested across the full socio

  17. Ecological Prose“The Cambridge I Know”by Xu Zhi-Mo and His Ecological Ideal%论徐志摩生态散文《我所知道的康桥》及其生态理想

    Institute of Scientific and Technical Information of China (English)

    郭茂全

    2013-01-01

      徐志摩《我所知道的康桥》是中国现代生态散文的代表作。作家以留学剑桥大学的生活体验为书写对象,描绘了一幅自然与人文和谐统一的校园生态画卷,表现了作家热爱自然、崇尚生命、向往自由、追求本真的生态理想。徐志摩生态观的形成不仅受到江南地域生态文化与中国传统生态观念的浸润,还受到受西方浪漫主义文学思潮与剑桥大学校园生态文化的滋养。%The Cambridge I know by XU Zhi-Mo is a masterpiece of modern Chinese ecological prose. Set on his life in University of Cambridge, the essay draws a picture of a campus where human is harmonious with nature, and expresses the writer’s ecological ideal for nature, life, freedom and inwardness. The ecological viewpoint of XU Zhi-Mo is not only affected by ecological culture in South China and traditional Chinese ecological concept, but also by ecological thought of the western romanticism literary trend and ecological culture of the University of Cambridge.

  18. Falls in advanced old age: recalled falls and prospective follow-up of over-90-year-olds in the Cambridge City over-75s Cohort study

    Directory of Open Access Journals (Sweden)

    Matthews Fiona E

    2008-03-01

    Full Text Available Abstract Background The "oldest old" are now the fastest growing section of most western populations, yet there are scarcely any data concerning even the common problem of falls amongst the very old. Prospective data collection is encouraged as the most reliable method for researching older people's falls, though in clinical practice guidelines advise taking a history of any recalled falls. This study set out to inform service planning by describing the epidemiology of falls in advanced old age using both retrospectively and prospectively collected falls data. Methods Design: Re-survey of over-90-year-olds in a longitudinal cohort study – cross-sectional interview and intensive 12-month follow-up. Participants and setting: 90 women and 20 men participating in a population-based cohort (aged 91–105 years, in care-homes and community-dwelling recruited from representative general practices in Cambridge, UK Measurements: Prospective falls data were collected using fall calendars and telephone follow-up for one year after cross-sectional survey including fall history. Results 58% were reported to have fallen at least once in the previous year and 60% in the 1-year follow-up. The proportion reported to have fallen more than once was lower using retrospective recall of the past year than prospective reports gathered the following year (34% versus 45%, as were fall rates (1.6 and 2.8 falls/person-year respectively. Repeated falls in the past year were more highly predictive of falls during the following year – IRR 4.7, 95% CI 2.6–8.7 – than just one – IRR 3.6, 95% CI 2.0–6.3, using negative binomial regression. Only 1/5 reportedly did not fall during either the year before or after interview. Conclusion Fall rates in this representative sample of over-90-year-olds are even higher than previous reports from octogenarians. Recalled falls last year, particularly repeated falls, strongly predicted falls during follow-up. Similar proportions

  19. Detection of classical and atypical/Nor98 scrapie by the paraffin-embedded tissue blot method.

    Science.gov (United States)

    Wemheuer, W M; Benestad, S L; Wrede, A; Wemheuer, W E; Brenig, B; Bratberg, B; Schulz-Schaeffer, W J

    2009-05-30

    The paraffin-embedded tissue (PET) blot method was used to investigate sections of the central nervous system and lymphatic tissues from 24 cases of classical scrapie and 25 cases of atypical/Nor98 scrapie in sheep and four healthy control sheep. The PET blot detected deposits of PrP(Sc) in the brain tissue of all 49 sheep with scrapie but no PrP(Sc) labelling could be detected in the control sheep. By contrast, not all the atypical/Nor98 scrapie cases were detectable by immunohistochemistry. The high sensitivity of the PET blot method made it possible to observe that in some atypical/Nor98 cases, deposits of PrP(Sc) may be restricted to supratentorial brain structures and that the diagnosis may be missed when only testing the obex area, where deposits are common in classical scrapie, and the cerebellar structures, where deposits are considered to be common in atypical/Nor98 cases.

  20. A slot blot immunoassay for quantitative detection of Plasmodium falciparum circumsporozoite protein in mosquito midgut oocyst.

    Directory of Open Access Journals (Sweden)

    Sanjai Kumar

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

  1. A teaching skills assessment tool inspired by the Calgary-Cambridge model and the patient-centered approach.

    Science.gov (United States)

    Sommer, Johanna; Lanier, Cédric; Perron, Noelle Junod; Nendaz, Mathieu; Clavet, Diane; Audétat, Marie-Claude

    2016-04-01

    The aim of this study was to develop a descriptive tool for peer review of clinical teaching skills. Two analogies framed our research: (1) between the patient-centered and the learner-centered approach; (2) between the structures of clinical encounters (Calgary-Cambridge communication model) and teaching sessions. During the course of one year, each step of the action research was carried out in collaboration with twelve clinical teachers from an outpatient general internal medicine clinic and with three experts in medical education. The content validation consisted of a literature review, expert opinion and the participatory research process. Interrater reliability was evaluated by three clinical teachers coding thirty audiotaped standardized learner-teacher interactions. This tool contains sixteen items covering the process and content of clinical supervisions. Descriptors define the expected teaching behaviors for three levels of competence. Interrater reliability was significant for eleven items (Kendall's coefficient pteaching skills. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

  2. International Organization of Standardization (ISO) and Cambridge Filter Test (CFT) Smoking Regimen Data Comparisons in Tobacco Product Marketing Applications.

    Science.gov (United States)

    Chae, Changyu; Walters, Matthew J; Holman, Matthew R

    2017-07-01

    We investigated the differences in TNCO (tar, nicotine, and carbon monoxide) smoke yields generated under the International Organization of Standardization (ISO) and Federal Trade Commission (FTC) Cambridge Filter Test (CFT) smoking regimens. Twenty-nine commercial cigarette products from the US marketplace were acquired in 2015 and tested by measuring the TNCO smoke yields generated under these 2 nonintense smoking regimens. Data obtained demonstrated a linear relationship between the TNCO yields produced under the 2 smoking regimens (R(2) > 0.99). TNCO yields produced by each product were higher under the CFT smoking regimen than the ISO smoking regimen. We found that tar, nicotine, and carbon monoxide yields were consistently 10% to 13% higher under the CFT smoking regimen than under the ISO smoking regimen. This strong correlation indicates that the 2 smoking regimens can be used to apply a correlation correction to CFT TNCO data and allow its comparison to ISO TNCO data in tobacco product marketing applications.

  3. A comparison of three infant skinfold reference standards: Tanner-Whitehouse, Cambridge Infant Growth Study, and WHO Child Growth Standards.

    Science.gov (United States)

    Miller, Elizabeth M

    2015-10-01

    As researchers increasingly focus on early infancy as a critical period of development, there is a greater need for methodological tools that can address all aspects of infant growth. Infant skinfold measures, in particular, are measurements in need of reliable reference standards that encompass all ages of infants and provide an accurate assessment of the relative fatness of a population. This report evaluates three published reference standards for infant skinfold measurements: Tanner-Whitehouse, Cambridge Infant Growth Study, and the World Health Organization (WHO) Child Growth Standards. To assess these standards, triceps skinfolds from a population of rural Kenyan infants (n = 250) and triceps skinfolds and subscapular skinfolds from infants in the National Health and Nutrition Examination Survey 1999-2002 (NHANES; n = 1197) were calculated as z-scores from the lambda-mu-sigma curves provided by each reference population. The Tanner-Whitehouse standards represented both the Kenyan and US populations as lean, while the Cambridge standards represented both populations as overfat. The distribution of z-scores based on the WHO standards fell in the middle, but excluded infants from both populations who were below the age of 3 months. Based on these results, the WHO reference standard is the best skinfold reference standard for infants over the age of 3 months. For populations with infants of all ages, the Tanner-Whitehouse standards are recommended, despite representing both study populations as underfat. Ideally, the WHO will extend their reference standard to include infants between the ages of 0 and 3 months.

  4. Anatomic variations of the pancreatic duct and their relevance with the Cambridge classification system: MRCP findings of 1158 consecutive patients.

    Science.gov (United States)

    Adibelli, Zehra Hilal; Adatepe, Mustafa; Imamoglu, Cetin; Esen, Ozgur Sipahi; Erkan, Nazif; Yildirim, Mehmet

    2016-12-01

    The study was conducted to evaluate the frequencies of the anatomic variations and the gender distributions of these variations of the pancreatic duct and their relevance with the Cambridge classification system as morphological sign of chronic pancreatitis using magnetic resonance cholangiopancreatography (MRCP). We retrospectively reviewed 1312 consecutive patients who referred to our department for MRCP between January 2013 and August 2015. We excluded 154 patients from the study because of less than optimal results due to imaging limitations or a history of surgery on pancreas. Finally a total of 1158 patients were included in the study. Among the 1158 patients included in the study, 54 (4.6%) patients showed pancreas divisum, 13 patients (1.2%) were defined as ansa pancreatica. When we evaluated the course of the pancreatic duct, we found the prevalence 62.5% for descending, 30% for sigmoid, 5.5% for vertical and 2% for loop. The most commonly observed pancreatic duct configuration was Type 3 in 528 patients (45.6%) where 521 patients (45%) had Type 1 configuration. Vertical course (p = 0.004) and Type 2 (p = 0.03) configuration of pancreatic duct were more frequent in females than males. There were no statistically significant differences between the gender for the other pancreatic duct variations such as pancreas divisium, ansa pancreatica and course types other than vertical course (p > 0.05 for all). Variants of pancreas divisum and normal pancreatic duct variants were not associated with morphologic findings of chronic pancreatitis by using the Cambridge classification system. The ansa pancreatica is a rare type of anatomical variation of the pancreatic duct, which might be considered as a predisposing factor to the onset of idiopathic pancreatitis.

  5. Hydrogen-bond landscapes, geometry and energetics of squaric acid and its mono- and dianions: a Cambridge Structural Database, IsoStar and computational study

    NARCIS (Netherlands)

    F.H. Allen; A.J. Cruz-Cabeza; P.A. Wood; D.A. Bardwell

    2013-01-01

    As part of a programme of work to extend central-group coverage in the Cambridge Crystallographic Data Centre's (CCDC) IsoStar knowledge base of intermolecular interactions, we have studied the hydrogen-bonding abilities of squaric acid (H(2)SQ) and its mono-and dianions (HSQ(-) and SQ(2-)) using th

  6. Decline of Executive Function in a Clinical Population: Age, Psychopathology, and Test Performance on the Cambridge Neuropsychological Test Automated Battery (CANTAB)

    NARCIS (Netherlands)

    Janssen, G.T.L.; Aken, L. van; Mey, H.R.A. De; Witteman, C.L.M.; Egger, J.I.M.

    2014-01-01

    This study presents a cross-sectional examination of the age-related executive changes in a sample of adults with a history of psychiatric illness using the Cambridge Neuropsychological Test Automated Battery. A total of 406 patients, aged 18 to 72 years old, completed executive function tests of wo

  7. Trajectories of Offending and Their Relation to Life Failure in Late Middle Age: Findings from the Cambridge Study in Delinquent Development

    Science.gov (United States)

    Piquero, Alex R.; Farrington, David P.; Nagin, Daniel S.; Moffitt, Terrie E.

    2010-01-01

    Researchers have hypothesized that over the life course, criminal offending varies with problems in other domains, including life failure and physical and mental health. To examine this issue, the authors use data from the Cambridge Study in Delinquent Development, a prospective longitudinal survey of 411 South London males first studied at age 8…

  8. Teaching Three-Dimensional Structural Chemistry Using Crystal Structure Databases. 4. Examples of Discovery-Based Learning Using the Complete Cambridge Structural Database

    Science.gov (United States)

    Battle, Gary M.; Allen, Frank H.; Ferrence, Gregory M.

    2011-01-01

    Parts 1 and 2 of this series described the educational value of experimental three-dimensional (3D) chemical structures determined by X-ray crystallography and retrieved from the crystallographic databases. In part 1, we described the information content of the Cambridge Structural Database (CSD) and discussed a representative teaching subset of…

  9. Teaching Three-Dimensional Structural Chemistry Using Crystal Structure Databases. 2. Teaching Units that Utilize an Interactive Web-Accessible Subset of the Cambridge Structural Database

    Science.gov (United States)

    Battle, Gary M.; Allen, Frank H.; Ferrence, Gregory M.

    2010-01-01

    A series of online interactive teaching units have been developed that illustrate the use of experimentally measured three-dimensional (3D) structures to teach fundamental chemistry concepts. The units integrate a 500-structure subset of the Cambridge Structural Database specially chosen for their pedagogical value. The units span a number of key…

  10. Teaching Three-Dimensional Structural Chemistry Using Crystal Structure Databases. 3. The Cambridge Structural Database System: Information Content and Access Software in Educational Applications

    Science.gov (United States)

    Battle, Gary M.; Allen, Frank H.; Ferrence, Gregory M.

    2011-01-01

    Parts 1 and 2 of this series described the educational value of experimental three-dimensional (3D) chemical structures determined by X-ray crystallography and retrieved from the crystallographic databases. In part 1, we described the information content of the Cambridge Structural Database (CSD) and discussed a representative teaching subset of…

  11. Performance on Cambridge Neuropsychological Test Automated Battery Subtests Sensitive to Frontal Lobe Function in People with Autistic Disorder: Evidence from the Collaborative Programs of Excellence in Autism Network

    Science.gov (United States)

    Ozonoff, Sally; Cook, Ian; Coon, Hilary; Dawson, Geraldine; Joseph, Robert M.; Klin, Ami; McMahon, William M.; Minshew, Nancy; Munson, Jeffrey A.

    2004-01-01

    Recent structural and functional imaging work, as well as neuropathology and neuropsychology studies, provide strong empirical support for the involvement of frontal cortex in autism. The Cambridge Neuropsychological Test Automated Battery (CANTAB) is a computer-administered set of neuropsychological tests developed to examine specific components…

  12. Hydrogen-bond landscapes, geometry and energetics of squaric acid and its mono- and dianions: a Cambridge Structural Database, IsoStar and computational study

    NARCIS (Netherlands)

    Allen, F.H.; Cruz-Cabeza, A.J.; Wood, P.A.; Bardwell, D.A.

    2013-01-01

    As part of a programme of work to extend central-group coverage in the Cambridge Crystallographic Data Centre's (CCDC) IsoStar knowledge base of intermolecular interactions, we have studied the hydrogen-bonding abilities of squaric acid (H(2)SQ) and its mono-and dianions (HSQ(-) and SQ(2-)) using

  13. Proceedings of the Aircraft Wake Vortices Conference, March 15-17, 1977, held at the Transportation Systems Center, Kendall Square, Cambridge, MA

    Science.gov (United States)

    1977-06-01

    CA. Jan. 1977. 309 -1.A IM ASSESSMENT OF ATMOSPHERIC EFFECTS ON THE BEHAVIOR OF AIRCRAFT WAKE VORTICES PAUL B. MACCREADY. JR. AND PETER B.S. LISSAMAN...Transportation Systems Center Civil Aviation Authority Kendall Square London Space House Cambridge MA 02142 Kingsway WC2 England Pitchford . Lynn D. Lockheed

  14. Update on the Status of the On-Going Range Dependent Low Frequency Active Sonar Model Benchmarking Effort : From Cambridge to Kos [abstract

    NARCIS (Netherlands)

    Zampolli, M.; Ainslie, M.A.

    2011-01-01

    In April 2010, a symposium in Memory of David Weston was held at Clare College in Cambridge (UK). International researchers from academia and research laboratories met to discuss two sets of test problems for sonar performance models, one aimed at understanding mammal echolocation sonar („Problem AI

  15. Comparative Coh-Metrix Analysis of Reading Comprehension Texts: Unified (Russian) State Exam in English vs. Cambridge First Certificate in English

    Science.gov (United States)

    Solnyshkina, Marina I.; Harkova, Elena V.; Kiselnikov, Aleksander S.

    2014-01-01

    The article summarizes the results of the comparative study of Reading comprehension texts used in B2 level tests: Unified (Russia) State Exam in English (EGE) and Cambridge First Certificate in English (FCE). The research conducted was mainly focused on six parameters measured with the Coh-Metrix, a computational tool producing indices of the…

  16. The structure of N-arylindazoles and their aza-derivatives in the solid state: A systematic analysis of the Cambridge Structural Database coupled with DFT calculations

    Science.gov (United States)

    Alkorta, Ibon; Elguero, José

    2017-06-01

    A search in the Cambridge Structural Database for N-aryl indazoles and their aza derivatives affords 227 structures (183 1-aryl and 44 2-aryl). To discuss their structures, DFT calculations on 20 model compounds were carried out. The geometry of the five-membered ring (the pyrazole) and the conformation if the N-aryl substituent were analyzed.

  17. Association of resistance to activated protein with the presence of Leiden and Cambridge Factor V mutations in Mexican patients with primary thrombophilia.

    Science.gov (United States)

    Zavala Hernández, César; Hernández Zamora, Edgar; Martínez Murillo, Carlos; Arenas Sordo, María de la Luz; González Orozco, Ana Elena; Reyes Maldonado, Elba

    2010-01-01

    Leiden and Cambridge factor V coagulation mutations and activated protein C resistance (RaPC) are alterations related with vein and artery thrombosis. In this study we aimed to determine whether RaPC is associated with the presence of Leiden and Cambridge mutation and the frequency of these mutations in the racially mestizo Mexican population. We included 150 Mexican patients with primary thrombophilia and 100 healthy subjects in this study. RaPC was determined using commercial methods and genotypes FV Leiden and FV Cambridge with PCR-RFLPs. RaPC was positive in four patients and in one control individual; however, there was no presence of Leiden or Cambridge mutation in the studied group; thus, RaPC was not correlated with the presence of any of the studied mutations. These results indicate that there are other primary or secondary causes different from those studied, which condition the presence of RaPC. Furthermore, the frequency obtained for RaPC in our thrombophilic population of racially mixed Mexicans is lower compared to that obtained in the Caucasian population, most probably because they are genetically different populations.

  18. Correlates of time spent walking and cycling to and from work: baseline results from the commuting and health in Cambridge study

    OpenAIRE

    Panter, J; Griffin, S.; Jones, A; MacKett, R; Ogilvie, D

    2011-01-01

    Environmental perceptions and psychological measures appear to be associated with walking and cycling behaviour; however, their influence is still unclear. We assessed these associations using baseline data from a quasi-experimental cohort study of the effects of major transport infrastructural developments in Cambridge, UK.

  19. 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...

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

    NARCIS (Netherlands)

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

    1996-01-01

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