WorldWideScience

Sample records for standard assay conditions

  1. Standardizing electrophoresis conditions: how to eliminate a major source of error in the comet assay.

    Directory of Open Access Journals (Sweden)

    Gunnar Brunborg

    2015-06-01

    Full Text Available In the alkaline comet assay, cells are embedded in agarose, lysed, and then subjected to further processing including electrophoresis at high pH (>13. We observed very large variations of mean comet tail lengths of cell samples from the same population when spread on a glass or plastic substrate and subjected to electrophoresis. These variations might be cancelled out if comets are scored randomly over a large surface, or if all the comets are scored. The mean tail length may then be representative of the population, although its standard error is large. However, the scoring process often involves selection of 50 – 100 comets in areas selected in an unsystematic way from a large gel on a glass slide. When using our 96-sample minigel format (1, neighbouring sample variations are easily detected. We have used this system to study the cause of the comet assay variations during electrophoresis and we have defined experimental conditions which reduce the variations to a minimum. We studied the importance of various physical parameters during electrophoresis: (i voltage; (ii duration of electrophoresis; (iii electric current; (iv temperature; and (v agarose concentration. We observed that the voltage (V/cm varied substantially during electrophoresis, even within a few millimetres of distance between gel samples. Not unexpectedly, both the potential ( V/cm and the time were linearly related to the mean comet tail, whereas the current was not. By measuring the local voltage with microelectrodes a few millimetres apart, we observed substantial local variations in V/cm, and they increased with time. This explains the large variations in neighbouring sample comet tails of 25% or more. By introducing simple technology (circulation of the solution during electrophoresis, and temperature control, these variations in mean comet tail were largely abolished, as were the V/cm variations. Circulation was shown to be particularly important and optimal conditions

  2. Standardization of portable assay instrumentation: the neutron-coincidence tree

    International Nuclear Information System (INIS)

    Menlove, H.O.

    1983-01-01

    Standardization of portable neutron assay instrumentation has been achieved by using the neutron coincidence technique as a common basis for a wide range of instruments and applications. The electronics originally developed for the High-Level Neutron Coincidence Counter has been adapted to both passive- and active-assay instrumentation for field verification of bulk plutonium, inventory samples, pellets, powders, nitrates, high-enriched uranium, and materials-testing-reactor, light-water-reactor, and mixed-oxide fuel assemblies. The family of detectors developed at Los Alamos National Laboratory and their performance under in-field conditions are described. 16 figures, 3 tables

  3. Measuring enzyme activities under standardized in vivo-like conditions for Systems Biology

    NARCIS (Netherlands)

    van Eunen, K.; Bouwman, J.; Daran-Lapujade, P.A.L.; Postmus, J.; Canelas, A.; Mensonides, F.I.C.; Orij, R.; Tuzun, I.; van der Brink, J.; Smits, G.J.; van Gulik, W.M.; Brul, S.; Heijnen, J.J.; de Winde, J.H.; Teixeira de Mattos, M.J.; Kettner, C.; Nielsen, J.; Westerhoff, H.V.; Bakker, B.M.

    2010-01-01

    Realistic quantitative models require data from many laboratories. Therefore, standardization of experimental systems and assay conditions is crucial. Moreover, standards should be representative of the in vivo conditions. However, most often, enzyme-kinetic parameters are measured under assay

  4. Measuring enzyme activities under standardized in vivo-like conditions for systems biology

    NARCIS (Netherlands)

    van Eunen, Karen; Bouwman, Jildau; Daran-Lapujade, Pascale; Postmus, Jarne; Canelas, Andre B.; Mensonides, Femke I. C.; Orij, Rick; Tuzun, Isil; van den Brink, Joost; Smits, Gertien J.; van Gulik, Walter M.; Brul, Stanley; de Winde, Johannes H.; de Mattos, M. J. Teixeira; Kettner, Carsten; Nielsen, Jens; Westerhoff, Hans V.; Bakker, Barbara M.; Heijnen, J.J.

    Realistic quantitative models require data from many laboratories. Therefore, standardization of experimental systems and assay conditions is crucial. Moreover, standards should be representative of the in vivo conditions. However, most often, enzyme-kinetic parameters are measured under assay

  5. The optimal condition of performing MTT assay for the determination of radiation sensitivity

    International Nuclear Information System (INIS)

    Hong, Semie; Kim, Il Han

    2001-01-01

    The measurement of radiation survival using a clonogenic assay, the established standard, can be difficult and time consuming. In this study, We have used the MTT assay, based on the reduction of a tetrazolium salt to a purple formazan precipitate by living cells, as a substitution for clonogenic assay and have examined the optimal condition for performing this assay in determination of radiation sensitivity. Four human cancer cell lines - PCI-1, SNU-1066, NCI-H63O and RKO cells have been used. For each cell line, a clonogenic assay and a MTT assay using Premix WST-1 solution, which is one of the tetrazolium salts and does not require washing or solubilization of the precipitate were carried out after irradiation of 0, 2, 4, 6, 8, 10 Gy, For clonogenic assay, cells in 25 cm 2 flasks were irradiated after overnight incubation and the resultant colonies containing more than 50 cells were scored after culturing the cells for 10-14 days, For MTT assay, the relationship between absorbance and cell number, optimal seeding cell number, and optimal timing of assay was determined. Then, MTT assay was performed when the irradiated cells had regained exponential growth or when the non-irradiated cells had undergone four or more doubling times. There was minimal variation in the values gained from these two methods with the standard deviation generally less than 5%, and there were no statistically significant differences between two methods according to t-test in low radiation dose (below 6 Gy). The regression analyses showed high linear correlation with the R 2 value of 0.975-0.992 between data from the two different methods. The optimal cell numbers for MTT assay were found to be dependent on plating efficiency of used cell line. Less than 300 cells/well were appropriate for cells with high plating efficiency (more than 30%). For cells with low plating efficiency (less than 30%), 500 cells/well or more were appropriate for assay. The optimal time for MTT assay was alter 6

  6. Standardization of Neisseria meningitidis Serogroup B Colorimetric Serum Bactericida Assay

    Science.gov (United States)

    Rodríguez, Tamara; Lastre, Miriam; Cedré, Barbara; Campo, Judith del; Bracho, Gustavo; Zayas, Caridad; Taboada, Carlos; Díaz, Miriam; Sierra, Gustavo; Pérez, Oliver

    2002-01-01

    The correlate of protection for serogroup B meningococci is not currently known, but for serogroup C it is believed to be the serum bactericidal assay (SBA). The current SBAs are labor intensive and the variations in protocols among different laboratories make interpretation of results difficult. A colorimetric SBA (cSBA), based on the ability of Neisseria meningitidis serogroup B to consume glucose, leading to acid production, was standardized by using group B strain Cu385-83 as the target. The cSBA results were compared to those obtained for a traditional colony-counting microassay (mSBA). Glucose and bromocresol purple pH indicator were added to the medium in order to estimate growth of cSBA target cell survivors through color change. Different variants of the assay parameters were optimized: growth of target cells (Mueller Hinton agar plates), target cell number (100 CFU/per well), and human complement source used at a final concentration of 25%. After the optimization, three other group B strains (H44/76, 490/91, and 511/91) were used as targets for the cSBA. The selection of the assay parameters and the standardization of cSBA were done with 13 sera from vaccinated volunteers. The titers were determined as the higher serum dilution that totally inhibited the bacterial growth marked by the color invariability of the pH indicator. This was detected visually as well as spectrophotometrically and was closely related to a significant difference in the growth of target cell survivors determined using Student’s t test. Intralaboratory reproducibility was ±1 dilution. The correlation between bactericidal median titers and specific immunoglobulin G serum concentration by enzyme immunoassay was high (r = 0.910, P < 0.01). The bactericidal titers generated by the cSBA and the mSBA were nearly identical, and there was a high correlation between the two assays (r = 0.974, P < 0.01). The standardized cSBA allows easy, fast, and efficient evaluation of samples. PMID

  7. Multicenter Evaluation of Cystatin C Measurement after Assay Standardization.

    Science.gov (United States)

    Bargnoux, Anne-Sophie; Piéroni, Laurence; Cristol, Jean-Paul; Kuster, Nils; Delanaye, Pierre; Carlier, Marie-Christine; Fellahi, Soraya; Boutten, Anne; Lombard, Christine; González-Antuña, Ana; Delatour, Vincent; Cavalier, Etienne

    2017-04-01

    Since 2010, a certified reference material ERM-DA471/IFCC has been available for cystatin C (CysC). This study aimed to assess the sources of uncertainty in results for clinical samples measured using standardized assays. This evaluation was performed in 2015 and involved 7 clinical laboratories located in France and Belgium. CysC was measured in a panel of 4 serum pools using 8 automated assays and a candidate isotope dilution mass spectrometry reference measurement procedure. Sources of uncertainty (imprecision and bias) were evaluated to calculate the relative expanded combined uncertainty for each CysC assay. Uncertainty was judged against the performance specifications derived from the biological variation model. Only Siemens reagents on the Siemens systems and, to a lesser extent, DiaSys reagents on the Cobas system, provided results that met the minimum performance criterion calculated according to the intraindividual and interindividual biological variations. Although the imprecision was acceptable for almost all assays, an increase in the bias with concentration was observed for Gentian reagents, and unacceptably high biases were observed for Abbott and Roche reagents on their own systems. This comprehensive picture of the market situation since the release of ERM-DA471/IFCC shows that bias remains the major component of the combined uncertainty because of possible problems associated with the implementation of traceability. Although some manufacturers have clearly improved their calibration protocols relative to ERM-DA471, most of them failed to meet the criteria for acceptable CysC measurements. © 2016 American Association for Clinical Chemistry.

  8. Conditional Dependence in Microbial Forensic Assays - A Primer

    Energy Technology Data Exchange (ETDEWEB)

    Velsko, Stephan P. [Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States)

    2013-11-08

    This report provides an introduction to the topic of conditional dependence in the context of microbial forensic assays. Conditional dependence between two items of evidence E1 and E2 occurs when they are both used to support a hypothesis, but E1 affects the probability of E2 and vice versa. Ignoring this dependence can lead to very large errors in estimating the diagnosticity of the combined evidence. To introduce readers to this concept, a number of definitions of conditional dependence that have been used by authors in the past have been collected together and compared. Formal mathematical relationships that constrain conditional dependence are summarized. There are several specific scenarios in which unrecognized conditional dependence can arise in microbial forensic contexts. This report provides some notional examples that illustrate dramatic effects of conditional dependence on the weight of microbial forensic evidence, and discusses the relevance of these observations for the validation of microbial forensic assays. A two-­parameter model that describes the transition between various limiting forms of conditional dependence relations is provided in an appendix.

  9. Experience with local lymph node assay performance standards using standard radioactivity and nonradioactive cell count measurements.

    Science.gov (United States)

    Basketter, David; Kolle, Susanne N; Schrage, Arnhild; Honarvar, Naveed; Gamer, Armin O; van Ravenzwaay, Bennard; Landsiedel, Robert

    2012-08-01

    The local lymph node assay (LLNA) is the preferred test for identification of skin-sensitizing substances by measuring radioactive thymidine incorporation into the lymph node. To facilitate acceptance of nonradioactive variants, validation authorities have published harmonized minimum performance standards (PS) that the alternative endpoint assay must meet. In the present work, these standards were applied to a variant of the LLNA based on lymph node cell counts (LNCC) run in parallel as a control with the standard LLNA with radioactivity measurements, with threshold concentrations (EC3) being determined for the sensitizers. Of the 22 PS chemicals tested in this study, 21 yielded the same results from standard radioactivity and cell count measurements; only 2-mercaptobenzothiazole was positive by LLNA but negative by LNCC. Of the 16 PS positives, 15 were positive by LLNA and 14 by LNCC; methylmethacrylate was not identified as sensitizer by either of the measurements. Two of the six PS negatives tested negative in our study by both LLNA and LNCC. Of the four PS negatives which were positive in our study, chlorobenzene and methyl salicylate were tested at higher concentrations than the published PS, whereas the corresponding concentrations resulted in consistent negative results. Methylmethacrylate and nickel chloride tested positive within the concentration range used for the published PS. The results indicate cell counts and radioactive measurements are in good accordance within the same LLNA using the 22 PS test substances. Comparisons with the published PS results may, however, require balanced analysis rather than a simple checklist approach. Copyright © 2011 John Wiley & Sons, Ltd.

  10. Standard guide for making quality nondestructive assay measurements

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2009-01-01

    1.1 This guide is a compendium of Quality Measurement Practices for performing measurements of radioactive material using nondestructive assay (NDA) instruments. The primary purpose of the guide is to assist users in arriving at quality NDA results, that is, results that satisfy the end user’s needs. This is accomplished by providing an acceptable and uniform basis for the collection, analysis, comparison, and application of data. The recommendations are not compulsory or prerequisites to achieving quality NDA measurements, but are considered contributory in most areas. 1.2 This guide applies to the use of NDA instrumentation for the measurement of nuclear materials by the observation of spontaneous or stimulated nuclear radiations, including photons, neutrons, or the flow of heat. Recommended calibration, operating, and assurance methods represent guiding principles based on current NDA technology. The diversity of industry-wide nuclear materials measurement applications and instrumentation precludes disc...

  11. Creatinine Assay Attainment of Analytical Performance Goals Following Implementation of IDMS Standardization

    Directory of Open Access Journals (Sweden)

    Elizabeth Sunmin Lee

    2017-02-01

    Full Text Available Background: The international initiative to standardize creatinine (Cr assays by tracing reference materials to Isotope Dilution Mass Spectrometry (IDMS assigned values was implemented to reduce interlaboratory variability and improve assay accuracy. Objective: The aims of this study were to examine whether IDMS standardization has improved Cr assay accuracy (bias, interlaboratory variability (precision, total error (TE, and attainment of recommended analytical performance goals. Methods: External Quality Assessment (EQA data (n = 66 challenge vials from Ontario, Canada, were analyzed. The bias, precision, TE, and the number of EQA challenge vials meeting performance goals were determined by assay manufacturer before (n = 32 and after (n = 34 IDMS implementation. Results: The challenge vials with the worst bias and precision were spiked with known common interfering substances (glucose and bilirubin. IDMS standardization improved assay bias (10.4%-1.6%, P < .001, but precision remained unchanged (5.0%-4.7%, P = .5 with performance goals not consistently being met. Precision and TE goals based on biologic variation were attained by only 29% to 69% and 32% to 62% of challenge vials. Conclusions: While IDMS standardization has improved Cr assay accuracy and thus reduced TE, significant interlaboratory variability remains. Contemporary Cr assays do not currently meet the standards required to allow for accurate and consistent estimated glomerular filtration rate assessment and chronic kidney disease diagnosis across laboratories. Further improvements in Cr assay performance are needed.

  12. Qualification of standard membrane-feeding assay with Plasmodium falciparum malaria and potential improvements for future assays.

    Directory of Open Access Journals (Sweden)

    Kazutoyo Miura

    Full Text Available Vaccines that interrupt malaria transmission are of increasing interest and a robust functional assay to measure this activity would promote their development by providing a biologically relevant means of evaluating potential vaccine candidates. Therefore, we aimed to qualify the standard membrane-feeding assay (SMFA. The assay measures the transmission-blocking activity of antibodies by feeding cultured P. falciparum gametocytes to Anopheles mosquitoes in the presence of the test antibodies and measuring subsequent mosquito infection. The International Conference on Harmonisation (ICH Harmonised Tripartite Guideline Q2(R1 details characteristics considered in assay validation. Of these characteristics, we decided to qualify the SMFA for Precision, Linearity, Range and Specificity. The transmission-blocking 4B7 monoclonal antibody was tested over 6 feeding experiments at several concentrations to determine four suitable concentrations that were tested in triplicate in the qualification experiments (3 additional feeds to evaluate Precision, Linearity and Range. For Specificity, 4B7 was tested in the presence of normal mouse IgG. We determined intra- and inter-assay variability of % inhibition of mean oocyst intensity at each concentration of 4B7 (lower concentrations showed higher variability. We also showed that % inhibition was dependent on 4B7 concentration and the activity is specific to 4B7. Since obtaining empirical data is time-consuming, we generated a model using data from all 9 feeds and simulated the effects of different parameters on final readouts to improve the assay procedure and analytical methods for future studies. For example, we estimated the effect of number of mosquitoes dissected on variability of % inhibition, and simulated the relationship between % inhibition in oocyst intensity and % inhibition of prevalence of infected mosquitos at different mean oocysts in the control. SMFA is one of the few biological assays used in

  13. Defining cell culture conditions to improve human norovirus infectivity assays

    Energy Technology Data Exchange (ETDEWEB)

    Straub, Tim M. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Hutchison, Janine R. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Bartholomew, Rachel A. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Valdez, Catherine O. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Valentine, Nancy B. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Dohnalkova, Alice [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Ozanich, Richard M. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States); Bruckner-Lea, Cindy J. [Pacific Northwest National Lab. (PNNL), Richland, WA (United States)

    2013-01-10

    Significant difficulties remain for determining whether human noroviruses (hNoV) recovered from water, food, and environmental samples are infectious. Three-dimensional tissue culture of human intestinal cells has shown promise in developing an infectivity assay, but reproducibility, even within a single laboratory, remains problematic. From the literature and our observations, we hypothesized that the common factors that leads to more reproducible hNoV infectivity in vitro requires that the cell line be 1) of human gastrointestinal origin, 2) expresses apical microvilli, and 3) be a positive secretor cell line. The C2BBe1 cell line, which is a brush-border producing clone of Caco-2, meets these three criteria. When challenged with Genogroup II viruses, we observed a 2 Log10 increase in viral RNA titer. A passage experiment with GII viruses showed evidence of the ability to propagate hNoV by both reverse transcription quantitative PCR (qRT-PCR) and microscopy. Using 3-D C2BBe1 cells improves reproducibility of the infectivity assay for hNoV, but the assay can still be variable. Two sources of variability include the cells themselves (mixed phenotypes of small and large intestine) and initial titer measurements using quantitative reverse transcription PCR (qRT-PCR) that measures all RNA vs. plaque assays that measure infectious virus.

  14. Inter-laboratory variation in DNA damage using a standard comet assay protocol

    DEFF Research Database (Denmark)

    Forchhammer, Lykke; Ersson, Clara; Loft, Steffen

    2012-01-01

    There are substantial inter-laboratory variations in the levels of DNA damage measured by the comet assay. The aim of this study was to investigate whether adherence to a standard comet assay protocol would reduce inter-laboratory variation in reported values of DNA damage. Fourteen laboratories ...

  15. The use of calibration standards and the correction for sample self-attenuation in gamma-ray nondestructive assay

    International Nuclear Information System (INIS)

    Parker, J.L.

    1986-11-01

    The efficient use of appropriate calibration standards and the correction for the attenuation of the gamma rays within an assay sample by the sample itself are two important and closely related subjects in gamma-ray nondestructive assay. Much research relating to those subjects has been done in the Nuclear Safeguards Research and Development program at the Los Alamos National Laboratory since 1970. This report brings together most of the significant results of that research. Also discussed are the nature of appropriate calibration standards and the necessary conditions on the composition, size, and shape of the samples to allow accurate assays. Procedures for determining the correction for the sample self-attenuation are described at length including both general principles and several specific useful cases. The most useful concept is that knowing the linear attenuation coefficient of the sample (which can usually be determined) and the size and shape of the sample and its position relative to the detector permits the computation of the correction factor for the self-attenuation. A major objective of the report is to explain how the procedures for determining the self-attenuation correction factor can be applied so that calibration standards can be entirely appropriate without being particularly similar, either physically or chemically, to the items to be assayed. This permits minimization of the number of standards required to assay items with a wide range of size, shape, and chemical composition

  16. Toward an international standard for PCR-based detection of food-borne thermotolerant Campylobacters: Assay development and analytical validation

    DEFF Research Database (Denmark)

    Lübeck, Peter Stephensen; Wolffs, P.; On, Stephen L.W.

    2003-01-01

    As part of a European research project (FOOD-PCR), we developed a standardized and robust PCR detection assay specific for the three most frequently reported food-borne pathogenic Campylobacter species, C. jejuni, C. coli, and C. lari. Fifteen published and unpublished PCR primers targeting the 16S...... carcass rinse, unlike both Taq DNA polymerase and DyNAzyme. Based on these results, Tth was selected as the most suitable enzyme for the assay. The standardized PCR test described shows potential for use in large-scale screening programs for food-borne Campylobacter species under the assay conditions....... The inclusivity and exclusivity were 100 and 97%, respectively. In an attempt to find a thermostable DNA polymerase more resistant than Taq to PCR inhibitors present in chicken samples, three DNA polymerases were evaluated. The DNA polymerase Tth was not inhibited at a concentration of 2% (vol/vol) chicken...

  17. Evaluation of the dot enzyme-linked immunosorbent assay in comparison with standard ELISA for the immunodiagnosis of human toxocariasis

    Directory of Open Access Journals (Sweden)

    Roldán William

    2006-01-01

    Full Text Available A dot enzyme-linked immunosorbent assay (dot-ELISA was standardized using excretory-secretory antigens of Toxocara canis for the rapid immunodiagnosis of human toxocariasis. Thirty patients with clinical signs of toxocariasis, 20 cases with other parasitic diseases, and 40 healthy subjects were tested. A total of 0.2 ng of antigen per dot, serum dilution of 1:160 and dilution conjugate of 1:1000 were found optimal. The sensitivity and specificity of the assay were 100 and 95%, respectively. Comparable sensitivity of dot-ELISA and the standard ELISA was obtained, but only 3 cross-reactions occurred in the dot-ELISA, compared with 6 in the standard ELISA. Dot-ELISA is simple to perform, rapid, and low cost. Large-scale screening studies should be done to evaluate its usefulness under field conditions.

  18. Consent Codes: Upholding Standard Data Use Conditions.

    Directory of Open Access Journals (Sweden)

    Stephanie O M Dyke

    2016-01-01

    Full Text Available A systematic way of recording data use conditions that are based on consent permissions as found in the datasets of the main public genome archives (NCBI dbGaP and EMBL-EBI/CRG EGA.

  19. Standardization of Assays That Detect Anti-Rubella Virus IgG Antibodies

    Science.gov (United States)

    Grangeot-Keros, Liliane; Vauloup-Fellous, Christelle

    2015-01-01

    SUMMARY Rubella virus usually causes a mild infection in humans but can cause congenital rubella syndrome (CRS). Vaccination programs have significantly decreased primary rubella virus infection and CRS; however, vaccinated individuals usually have lower levels of rubella virus IgG than those with natural infections. Rubella virus IgG is quantified with enzyme immunoassays that have been calibrated against the World Health Organization (WHO) international standard and report results in international units per milliliter. It is recognized that the results reported by these assays are not standardized. This investigation into the reasons for the lack of standardization found that the current WHO international standard (RUB-1-94) fails by three key metrological principles. The standard is not a pure analyte but is composed of pooled human immunoglobulin. It was not calibrated by certified reference methods; rather, superseded tests were used. Finally, no measurement uncertainty estimations have been provided. There is an analytical and clinical consequence to the lack of standardization of rubella virus IgG assays, which leads to misinterpretation of results. The current approach to standardization of rubella virus IgG assays has not achieved the desired results. A new approach is required. PMID:26607813

  20. Measuring Cytotoxicity by Bioluminescence Imaging Outperforms the Standard Chromium-51 Release Assay

    Science.gov (United States)

    Karimi, Mobin A.; Lee, Eric; Bachmann, Michael H.; Salicioni, Ana Maria; Behrens, Edward M.; Kambayashi, Taku; Baldwin, Cynthia L.

    2014-01-01

    The chromium-release assay developed in 1968 is still the most commonly used method to measure cytotoxicity by T cells and by natural killer cells. Target cells are loaded in vitro with radioactive chromium and lysis is determined by measuring chromium in the supernatant released by dying cells. Since then, alternative methods have been developed using different markers of target cell viability that do not involve radioactivity. Here, we compared and contrasted a bioluminescence (BLI)-based cytotoxicity assay to the standard radioactive chromium-release assay using an identical set of effector cells and tumor target cells. For this, we stably transduced several human and murine tumor cell lines to express luciferase. When co-cultured with cytotoxic effector cells, highly reproducible decreases in BLI were seen in an effector to target cell dose-dependent manner. When compared to results obtained from the chromium release assay, the performance of the BLI-based assay was superior, because of its robustness, increased signal-to-noise ratio, and faster kinetics. The reduced/delayed detection of cytotoxicity by the chromium release method was attributable to the association of chromium with structural components of the cell, which are released quickly by detergent solubilization but not by hypotonic lysis. We conclude that the (BLI)-based measurement of cytotoxicity offers a superior non-radioactive alternative to the chromium-release assay that is more robust and quicker to perform. PMID:24586714

  1. Urolithins display both antioxidant and pro-oxidant activities depending on assay system and conditions.

    Science.gov (United States)

    Kallio, Tuija; Kallio, Johanna; Jaakkola, Mari; Mäki, Marianne; Kilpeläinen, Pekka; Virtanen, Vesa

    2013-11-13

    The biological effects of polyphenolic ellagitannins are mediated by their intestinal metabolites, urolithins. This study investigated redox properties of urolithins A and B using ORAC assay, three cell-based assays, copper-initiated pro-oxidant activity (CIPA) assay, and cyclic voltammetry. Urolithins were strong antioxidants in the ORAC assay, but mostly pro-oxidants in cell-based assays, although urolithin A was an antioxidant in cell culture medium. Parent compound ellagic acid was a strong extracellular antioxidant, but showed no response in the intracellular assay. The CIPA assay confirmed the pro-oxidant activity of ellagitannin metabolites. In the cell proliferation assay, urolithins but not ellagic acid decreased growth and metabolism of HepG2 liver cells. In cyclic voltammetry, the oxidation of urolithin A was partly reversible, but that of urolithin B was irreversible. These results illustrate how strongly measured redox properties depend on the employed assay system and conditions and emphasize the importance of studying pro-oxidant and antioxidant activities in parallel.

  2. Detection of systemic hypersensitivity to drugs using standard guinea pig assays.

    Science.gov (United States)

    Weaver, James L; Staten, David; Swann, Joslyn; Armstrong, George; Bates, Melissa; Hastings, Kenneth L

    2003-12-01

    The most commonly used assays designed to detect either skin or systemic immune-based hypersensitivity reactions are those using guinea pigs (GP). We obtained data from various FDA records to evaluate the correlation between GP assay results and reported post-marketing systemic hypersensitivity reactions. We examined the new drug application (NDA) reviews of approved drugs for the results of GP assays. Post-marketing human data were extracted from the FDA adverse event reporting system (AERS). Drug usage data were obtained from a commercial database maintained by IMS Health Inc. We found 83 (21%) of 396 drugs approved between 1978 and 1998 had reported GP test results. Among these 83 drugs, 14 (17%) were found to have positive results in at least one GP assay. Simple reporting index (RI) values for systemic hypersensitivity reactions were calculated from AERS data and usage to produce the index of adverse event reports per million shipping units of drug. A variety of definitions of positive human response were examined. A statistically significant association was seen for rash between post-marketing and clinical trials adverse event reports. No statistically significant associations between human data and GP test results were observed. These data suggest that standard GP assays have limited ability to predict human systemic hypersensitivity potential for pharmaceuticals.

  3. Evaluation of standard reagents for radial-immunodiffusion assays. In vitro control of rabies vaccines

    Directory of Open Access Journals (Sweden)

    MICELI Graciela S.

    2000-01-01

    Full Text Available The RID assay is one of the in vitro methods used for in-process control in the production of rabies vaccines for veterinary use. It has been shown to be very useful for determining antigen concentration in the final bulk product. The work presented in this paper, including the production and standardization of candidate standard reagents for use in the Radial Immunodiffusion Assay (RID was carried out at the Pan American Institute for Food Protection and Zoonoses (INPPAZ/PAHO/WHO. The study was completed with the cooperation of the Faculty of Veterinary Sciences, National University of La Plata (NULP, Argentina, where the validation of the proposed standards and the quality control of samples from 28 different batches of rabies vaccines produced with Pasteur strain rabies virus (PV in BHK cells were performed. The activity of the vaccines was determined by in vivo (NIH and in vitro (RIDassays. The results of the candidate reagents for the reagent standardization tests showed stability, sensitivity and reproducibility. The Relative Potency the 1.2 between the problem vaccines and the reference vaccine was estimated by variance and regression analysis. The results of our validation study show that the INPPAZ (PAHO/WHO is capable of producing and distributing the above-mentioned standard reagents, as well as of providing support for the incorporation of the RID technique (sensitive, rapid and inexpensive to the laboratories that manufacture rabies vaccines in Latin America and the Caribbean.

  4. The network formation assay: a spatially standardized neurite outgrowth analytical display for neurotoxicity screening.

    Science.gov (United States)

    Frimat, Jean-Philippe; Sisnaiske, Julia; Subbiah, Subanatarajan; Menne, Heike; Godoy, Patricio; Lampen, Peter; Leist, Marcel; Franzke, Joachim; Hengstler, Jan G; van Thriel, Christoph; West, Jonathan

    2010-03-21

    We present a rapid, reproducible and sensitive neurotoxicity testing platform that combines the benefits of neurite outgrowth analysis with cell patterning. This approach involves patterning neuronal cells within a hexagonal array to standardize the distance between neighbouring cellular nodes, and thereby standardize the length of the neurite interconnections. This feature coupled with defined assay coordinates provides a streamlined display for rapid and sensitive analysis. We have termed this the network formation assay (NFA). To demonstrate the assay we have used a novel cell patterning technique involving thin film poly(dimethylsiloxane) (PDMS) microcontact printing. Differentiated human SH-SY5Y neuroblastoma cells colonized the array with high efficiency, reliably producing pattern occupancies above 70%. The neuronal array surface supported neurite outgrowth, resulting in the formation of an interconnected neuronal network. Exposure to acrylamide, a neurotoxic reference compound, inhibited network formation. A dose-response curve from the NFA was used to determine a 20% network inhibition (NI(20)) value of 260 microM. This concentration was approximately 10-fold lower than the value produced by a routine cell viability assay, and demonstrates that the NFA can distinguish network formation inhibitory effects from gross cytotoxic effects. Inhibition of the mitogen-activated protein kinase (MAPK) ERK1/2 and phosphoinositide-3-kinase (PI-3K) signaling pathways also produced a dose-dependent reduction in network formation at non-cytotoxic concentrations. To further refine the assay a simulation was developed to manage the impact of pattern occupancy variations on network formation probability. Together these developments and demonstrations highlight the potential of the NFA to meet the demands of high-throughput applications in neurotoxicology and neurodevelopmental biology.

  5. Variability of assay methods for total and free PSA after WHO standardization.

    Science.gov (United States)

    Foj, L; Filella, X; Alcover, J; Augé, J M; Escudero, J M; Molina, R

    2014-03-01

    The variability of total PSA (tPSA) and free PSA (fPSA) results among commercial assays has been suggested to be decreased by calibration to World Health Organization (WHO) reference materials. To characterize the current situation, it is necessary to know its impact in the critical cutoffs used in clinical practice. In the present study, we tested 167 samples with tPSA concentrations of 0 to 20 μg/L using seven PSA and six fPSA commercial assays, including Access, ARCHITECT i2000, ADVIA Centaur XP, IMMULITE 2000, Elecsys, and Lumipulse G1200, in which we only measured tPSA. tPSA and fPSA were measured in Access using the Hybritech and WHO calibrators. Passing-Bablok analysis was performed for PSA, and percentage of fPSA with the Hybritech-calibrated access comparison assay. For tPSA, relative differences were more than 10 % at 0.2 μg/L for ARCHITECT i2000, and at a critical concentration of 3, 4, and 10 μg/L, the relative difference was exceeded by ADVIA Centaur XP and WHO-calibrated Access. For percent fPSA, at a critical concentration of 10 %, the 10 % relative difference limit was exceeded by IMMULITE 2000 assay. At a critical concentration of 20 and 25 %, ADVIA Centaur XP, ARCHITECT i2000, and IMMULITE 2000 assays exceeded the 10 % relative difference limit. We have shown significant discordances between assays included in this study despite advances in standardization conducted in the last years. Further harmonization efforts are required in order to obtain a complete clinical concordance.

  6. Reference assays for Clostridium difficile infection: one or two gold standards?

    Science.gov (United States)

    Planche, Timothy; Wilcox, Mark

    2011-01-01

    Accurate diagnosis of Clostridium difficile infection (CDI) is essential for optimal treatment, prevention and control. There are two reference assays for CDI diagnosis: the cell cytotoxicity assay (CCTA) and toxigenic culture (TC). Importantly, these tests actually detect different targets: CCTA detects the presence of C difficile toxins (primarily toxin B, but also toxin A), whereas TC detects the presence in the stool of C difficile with the potential to produce toxin. Not surprisingly studies comparing the results of these assays show imperfect agreement. Thus, a faecal sample may be CCTA negative but TC positive, and this raises the crucial question about the clinical significance of the presence of C difficile with the capacity to produce toxin but no actual detectable free toxin. A positive TC result indicates that a patient with diarrhoea is potentially infectious. TC also has the advantage that the cultured isolate is available for typing and for susceptibility testing. In general, however, CCTA has been shown to be a better test for the laboratory confirmation of CDI, although additional culture may be needed to optimise sensitivity. Crucially, when these reference assays are used to determine the accuracy of alternative diagnostic tests, care should be taken to compare methods with their appropriate standard (ie, compare tests that target equivalent end-points). Such issues have contributed to the variable and often suboptimal performance of rapid diagnostic tests for CDI. Further research is urgently needed to improve knowledge of the utility of routine diagnostic tests in CDI and the factors that influence their performance.

  7. Development of recombinant human IgA for anticardiolipin antibodies assay standardization.

    Science.gov (United States)

    Knappik, Achim; Capuano, Francesco; Frisch, Christian; Ylera, Francisco; Bonelli, Fabrizio

    2009-09-01

    Controls and calibrators in autoimmune assays are typically developed from patient sera. However, the use of sera is accompanied by a number of disadvantages, such as lack of monospecificity, lack of assay comparability, and supply limitations. Ideally, the control reagent would be an antigen-specific human monoclonal antibody preparation that is defined and pure, easy to produce without any supply limitations, and of defined isotype (IgG, IgM, or IgA). The generation of antigen-specific human monoclonal antibodies has been complicated, but recent advances in development of fully human antibodies by means of in vitro antibody gene library selection has opened a way for the isolation of human antibodies to virtually any antigen, including self-antigens. Such antibodies can be converted to any isotype by gene cloning. Here we developed a set of human monoclonal IgA antibodies specific for the cardiolipin-beta2-glycoprotein 1 complex, using the HuCAL technology. We evaluated the IgA variants of those antibodies for their use as standards in IgA anticardiolipin antibody assays and compared these reagents with serum controls. Such recombinant antibodies may ultimately replace patient sera as assay control and calibration reagents.

  8. 42 CFR 486.108 - Condition for coverage: Safety standards.

    Science.gov (United States)

    2010-10-01

    ... indication of the production of X-rays whenever the X-ray tube is energized. The control panel includes... BY SUPPLIERS Conditions for Coverage: Portable X-Ray Services § 486.108 Condition for coverage: Safety standards. X-ray examinations are conducted through the use of equipment which is free of...

  9. Guide to nondestructive assay standards: Preparation criteria, availability, and practical considerations

    International Nuclear Information System (INIS)

    Hsue, S.T.; Stewart, J.E.; Sampson, T.E.; Butler, G.W.; Rudy, C.R.; Rinard, P.M.

    1997-10-01

    For certification and measurement control, nondestructive assay (NDA) instruments and methods used for verification measurements of special nuclear materials (SNMs) require calibrations based on certified reference materials (CRMs), or working reference materials (WRMs), traceable to the national system of measurements, and adequately characteristic of the unknowns. The Department of Energy Office of Safeguards and Security is sponsoring production of a comprehensive guide to preparation of NDA standards. The scope of the report includes preparation criteria, current availability of CRMs and WRMs, practical considerations for preparation and characterization, and an extensive bibliography. In preparing the report, based primarily on experience at Los Alamos, they have found that standards preparation is highly dependent on the particular NDA method being applied. They therefore include sections that contain information specific to commonly used neutron and gamma-ray NDA techniques. They also present approaches that are alternatives to, or minimize requirements for physical standards

  10. Guide to nondestructive assay standards: Preparation criteria, availability, and practical considerations

    Energy Technology Data Exchange (ETDEWEB)

    Hsue, S.T.; Stewart, J.E.; Sampson, T.E.; Butler, G.W.; Rudy, C.R.; Rinard, P.M.

    1997-10-01

    For certification and measurement control, nondestructive assay (NDA) instruments and methods used for verification measurements of special nuclear materials (SNMs) require calibrations based on certified reference materials (CRMs), or working reference materials (WRMs), traceable to the national system of measurements, and adequately characteristic of the unknowns. The Department of Energy Office of Safeguards and Security is sponsoring production of a comprehensive guide to preparation of NDA standards. The scope of the report includes preparation criteria, current availability of CRMs and WRMs, practical considerations for preparation and characterization, and an extensive bibliography. In preparing the report, based primarily on experience at Los Alamos, they have found that standards preparation is highly dependent on the particular NDA method being applied. They therefore include sections that contain information specific to commonly used neutron and gamma-ray NDA techniques. They also present approaches that are alternatives to, or minimize requirements for physical standards.

  11. Reevaluation of the Harboe assay as a standardized method of assessment for the hemolytic performance of ventricular assist devices.

    Science.gov (United States)

    Chan, Chris H H; Hilton, Andrew; Foster, Graham; Hawkins, Karl

    2012-08-01

    The Harboe spectrophotometric assay is regarded as one of the safest and most reproducible methods for measuring plasma free hemoglobin (pfHb). However, there is still some ambiguity in the application of the assay when assessing the hemolytic performance of ventricular assist devices (VADs). The purpose of this study was to reexamine and compare values of pfHb obtained using different concentrations of plasma diluent (Na(2) CO(3) ) as cited by various studies such that a standard practice may be recommended for the application of the Harboe assay in the hemolytic evaluation of VADs, allowing reliable comparisons to be made between laboratories. As a means to examine the Harboe assay, a BioMedicus BPX-80 was tested using both whole blood and a washed suspension of red blood cells (RBCs). Results show that for whole blood, the pfHb may be underestimated by 13-23%, dependent upon the concentration of Na(2) CO(3) diluent solution. This trend was not observed for the washed suspension of RBCs. Furthermore, it is shown that the concentration of diluent influences the stability of a sample. The results of this study show that the problems associated with the incongruity of pfHb readings are a direct result of the precipitation of proteins from the plasma under alkaline conditions; as the molarity of the diluent controls pH, it becomes essential to use the appropriate concentration of Na(2) CO(3) diluent in order to avoid turbidity of the solution and the consequent misrepresentation of pfHb values. Such standardization is pertinent when measuring the very low levels of pfHb observed during the in vivo testing of modern ventricular assist devices. © 2012, Copyright the Authors. Artificial Organs © 2012, International Center for Artificial Organs and Transplantation and Wiley Periodicals, Inc.

  12. Study of the optimal reaction conditions for assay of the mouse alternative complement pathway

    NARCIS (Netherlands)

    Dijk, H. van; Rademaker, P.M.; Klerx, J.P.A.M.; Willers, J.M.M.

    1985-01-01

    The optimal reaction conditions for hemolytic assay of alternative complement pathway activity in mouse serum were investigated. A microtiter system was used, in which a number of 7.5×106 rabbit erythrocytes per test well appeared to be optimal. Rabbit erythrocytes were superior as target cells over

  13. Important Considerations for Methemoglobin Measurement in Fish Blood: Assay Choice and Storage Conditions

    Energy Technology Data Exchange (ETDEWEB)

    Kuntz, Mel Anton; Rodnick, K. J.; J. A. Lacey

    2002-05-01

    Spectrophotometric assays of methaemoglobin (metHb) in rainbow trout Oncorhynchus mykiss, channel catfish Ictalurus punctatus, tilapias Tilapia niloticus and Tilapia zillii and white sturgeon Acipenser transmontanus, under baseline conditions, were low (<4%) for each species, and yet higher than human values (<1%). MetHb results for a given fish species varied significantly between assays and two assays were deemed unacceptable for particular animals. For rainbow trout, white sturgeon, and the two species of tilapia, the Dubowski method gave uncharacteristically high estimates of metHb. MetHb could not measured in tilapia blood using the Evelyn & Malloy method due to spectral interference. Only the Horecker & Brackett assay worked well for all species. Storage conditions were extremely important in the quantification of metHb in rainbow trout blood. For consistent values, samples can be stored up to 4 h on ice (0 degrees C) or at least 20 days under liquid nitrogen (-196 degrees C). Auto-oxidation, however, elevates rainbow trout metHb at -20 and -80 degrees C. It should not be assumed that the blood of fishes and humans perform similarly during assays of metHb.

  14. Use of new membrane-reactor saccharification assay to evaluate the performance of cellulases under simulated SSF conditions

    Energy Technology Data Exchange (ETDEWEB)

    Baker, J.O.; Vinzant, T.B.; Ehrman, C.I. [National Renewable Energy Lab., Golden, CO (United States)] [and others

    1997-12-31

    A new saccharification assay has been devised, in which a continuously buffer-swept membrane reactor is used to remove the solubilized saccharification products, thus allowing high extents of substrate conversion without significant inhibitory effects from the buildup of either cellobiose or glucose. This diafiltration saccharification assay (DSA) can, therefore, be used to obtain direct measurements of the performance of combinations of cellulose and substrate under simulated SSF conditions, without the saccharification results being complicated by factors that may influence the subsequent fermentation step. This assay has been used to compare the effectiveness of commercial and special in-house-produced Trichoderma reesei cellulose preparations in the saccharification of a standardized microcrystalline (Sigmacell) substrate and a dilute-acid pretreated lignocellulosic substrate. Initial results strongly suggest that enzyme preparations produced in the presence of the targeted lignocellulosic substrate will saccharify that substrate more effectively. These results call into question the widespread use of the {open_quotes}filter paper assay{close_quotes} as a reliable predictor of enzyme performance in the extensive hydrolysis of substrates that are quite different from filter paper in both physical properties and chemical composition. 14 refs., 6 figs.

  15. Standard Model Vacuum Stability and Weyl Consistency Conditions

    DEFF Research Database (Denmark)

    Antipin, Oleg; Gillioz, Marc; Krog, Jens

    2013-01-01

    At high energy the standard model possesses conformal symmetry at the classical level. This is reflected at the quantum level by relations between the different beta functions of the model. These relations are known as the Weyl consistency conditions. We show that it is possible to satisfy them...... order by order in perturbation theory, provided that a suitable coupling constant counting scheme is used. As a direct phenomenological application, we study the stability of the standard model vacuum at high energies and compare with previous computations violating the Weyl consistency conditions....

  16. Standard test method for nondestructive assay of radioactive material by tomographic gamma scanning

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2010-01-01

    1.1 This test method describes the nondestructive assay (NDA) of gamma ray emitting radionuclides inside containers using tomographic gamma scanning (TGS). High resolution gamma ray spectroscopy is used to detect and quantify the radionuclides of interest. The attenuation of an external gamma ray transmission source is used to correct the measurement of the emission gamma rays from radionuclides to arrive at a quantitative determination of the radionuclides present in the item. 1.2 The TGS technique covered by the test method may be used to assay scrap or waste material in cans or drums in the 1 to 500 litre volume range. Other items may be assayed as well. 1.3 The test method will cover two implementations of the TGS procedure: (1) Isotope Specific Calibration that uses standards of known radionuclide masses (or activities) to determine system response in a mass (or activity) versus corrected count rate calibration, that applies to only those specific radionuclides for which it is calibrated, and (2) Respo...

  17. Guide to nondestructive assay standards: Preparation criteria, availability, and practical considerations

    International Nuclear Information System (INIS)

    Stewart, J.E.; Hsue, S.T.; Sampson, T.E.

    1997-01-01

    For certification and measurement control, nondestructive assay (NDA) instruments and methods used for verification measurement of special nuclear materials (SNMs) require calibrations based on certified reference materials (CRMs), or working reference materials (WRMs), traceable to the national system of measurements, and adequately characteristic of the unknowns. The Department of Energy Office of Safeguards and Security is sponsoring production of a comprehensive guide to preparation of NDA standards. The scope of the report includes preparation criteria, current availability of CRMs and WRMs, practical considerations for preparation and characterization, and an extensive bibliography. In preparing the report, based primarily on experience at Los Alamos, we have found that standards preparation is highly dependent on the particular NDA method being applied. We therefore include sections that contain information specific to commonly used neutron and gamma-ray NDA techniques. 16 refs., 4 figs., 2 tabs

  18. A single-centre evaluation of two new anti-Mullerian hormone assays and comparison with the current clinical standard assay.

    Science.gov (United States)

    Welsh, Paul; Smith, Karen; Nelson, Scott M

    2014-05-01

    .0%, P Gen II assay and may be suitable for clinical and epidemiological use. Enhanced sensitivity of the Ansh Labs picoAMH assay enables measurement of low AMH concentrations. These results re-emphasize the need for an AMH international standard. Ansh Labs provided kits for this study free of charge. The manufacturer played no part in conducting assays or data analysis. S.M.N. has received speaker's fees and participated in advisory boards for Beckman Coulter, Merck Serono, MSD and Ferring regarding AMH. P.W. is supported by British Heart Foundation fellowship FS/12/62/29889. We declare no other financial relationship or competing interests.

  19. Standardization of automated 25-hydroxyvitamin D assays: How successful is it?

    Science.gov (United States)

    Elsenberg, E H A M; Ten Boekel, E; Huijgen, H; Heijboer, A C

    2017-12-01

    Multiple 25(OH)D assays have recently been aligned to improve comparibility. In this study we investigated the performance of these assays using both native single-donor sera with target values certified by a reference method as well as single donor sera from a heterogeneous patient population. 25(OH)D levels were measured in twenty reference samples (Ref!25OHD; Labquality, Finland) using five automated methods (Lumipulse, Liaison, Cobas, iSYS and Access) and one aligned ID-XLC-MS/MS method (slope: 1,00; intercept: 0,00; R=0,996). Furthermore, 25(OH)D concentrations measured in 50 pregnant women and 52 random patients using the 5 automated assays were compared to the ID-XLC-MS/MS. In addition, Vitamin D binding protein (DBP) was measured. Most automated assays showed significant differences in 25(OH)D levels measured in reference samples. Slopes varied from 1,00 to 1,33, intercepts from -5.48 to -15,81nmol/L and the R from 0,971 to 0,997. This inaccuracy was even more prominent in a heterogeneous patient population. Slopes varied from 0,75 to 1,35, intercepts from -9.02 to 11,51nmol/L and the R from 0,840 to 0,949. For most assays the deviation in 25(OH)D concentration increased with elevating DBP concentrations suggesting that DBP might be one of the factors contributing to the inaccuracy in currently used automated 25(OH)D methods. Despite the use of standardized assays, we observed significant differences in 25(OH)D concentrations in some automated methods using reference material obtained from healthy single donor sera. In sera of a patient population this inaccuracy was even worse which is highly concerning as patient samples are being investigated in clinical laboratories. Copyright © 2017 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  20. Conditional Standard Errors of Measurement for Scale Scores.

    Science.gov (United States)

    Kolen, Michael J.; And Others

    1992-01-01

    A procedure is described for estimating the reliability and conditional standard errors of measurement of scale scores incorporating the discrete transformation of raw scores to scale scores. The method is illustrated using a strong true score model, and practical applications are described. (SLD)

  1. Conditional Wegner Estimate for the Standard Random Breather Potential

    Science.gov (United States)

    Täufer, Matthias; Veselić, Ivan

    2015-11-01

    We prove a conditional Wegner estimate for Schrödinger operators with random potentials of breather type. More precisely, we reduce the proof of the Wegner estimate to a scale free unique continuation principle. The relevance of such unique continuation principles has been emphasized in previous papers, in particular in recent years. We consider the standard breather model, meaning that the single site potential is the characteristic function of a ball or a cube. While our methods work for a substantially larger class of random breather potentials, we discuss in this particular paper only the standard model in order to make the arguments and ideas easily accessible.

  2. Assessing the cleanliness of surfaces: Innovative molecular approaches vs. standard spore assays

    Energy Technology Data Exchange (ETDEWEB)

    Cooper, M.; Duc, M.T. La; Probst, A.; Vaishampayan, P.; Stam, C.; Benardini, J.N.; Piceno, Y.M.; Andersen, G.L.; Venkateswaran, K.

    2011-04-01

    A bacterial spore assay and a molecular DNA microarray method were compared for their ability to assess relative cleanliness in the context of bacterial abundance and diversity on spacecraft surfaces. Colony counts derived from the NASA standard spore assay were extremely low for spacecraft surfaces. However, the PhyloChip generation 3 (G3) DNA microarray resolved the genetic signatures of a highly diverse suite of microorganisms in the very same sample set. Samples completely devoid of cultivable spores were shown to harbor the DNA of more than 100 distinct microbial phylotypes. Furthermore, samples with higher numbers of cultivable spores did not necessarily give rise to a greater microbial diversity upon analysis with the DNA microarray. The findings of this study clearly demonstrated that there is not a statistically significant correlation between the cultivable spore counts obtained from a sample and the degree of bacterial diversity present. Based on these results, it can be stated that validated state-of-the-art molecular techniques, such as DNA microarrays, can be utilized in parallel with classical culture-based methods to further describe the cleanliness of spacecraft surfaces.

  3. Standardization of CalyculinA induced PCC assay and its advantages over Okadaic acid PCC assay in Biodosimetry applications.

    Science.gov (United States)

    Nairy, Rajesha K; Yerol, Narayana; Bhat, Nagesh N; Desai, Utkarsha; Shirsath, Kapil; Yadav, Usha; K Chaurasia, Rajesh; B K, Sapra

    2016-11-29

    In the present study an attempt was made to estimate coefficients of dose response curves for PCC aberrations induced by CalyculinA and Okadaic acid, using 60 Co gamma radiation and 8 MeV pulsed electron beam for biodosimetry application. The modified method outlined by Puig et al. 2013 was used to conduct Calyculin A and Okadaic acid induced PCC assay in human blood lymphocytes.Chemical treatment was given for the last 1 h of a 48 h culture. The study was carried out in the dose range 2.5 to 20 Gy using 60 Co gamma rays and 8 MeV pulsed electron beam. Results show a linear dose dependent increase with a slope of 0.047 ± 0.001 from Calycalin A PCC and 0.048 ± 0.002 form Okadaic acid PCC. The slope of the fragments curve was 0.327 ± 0.006 from Calyculin A and 0.328 ± 0.006 from Okadaic acid PCC. Further, dose calibration studies were carried out for 8 MeV electron using Calyculin A PCC assay and the obtained slope from ring yield was 0.054 ± 0.002 and 0.427 ± 0.009 from fragment yield.

  4. Standardization of a Continuous Assay for Glycosidases and Its Use for Screening Insect Gut Samples at Individual and Populational Levels

    Directory of Open Access Journals (Sweden)

    Gerson S. Profeta

    2017-05-01

    Full Text Available Glycoside Hydrolases (GHs are enzymes able to recognize and cleave glycosidic bonds. Insect GHs play decisive roles in digestion, in plant-herbivore, and host-pathogen interactions. GH activity is normally measured by the detection of a release from the substrate of products as sugars units, colored, or fluorescent groups. In most cases, the conditions for product release and detection differ, resulting in discontinuous assays. The current protocols result in using large amounts of reaction mixtures for the obtainment of time points in each experimental replica. These procedures restrain the analysis of biological materials with limited amounts of protein and, in the case of studies regarding small insects, implies in the pooling of samples from several individuals. In this respect, most studies do not assess the variability of GH activities across the population of individuals from the same species. The aim of this work is to approach this technical problem and have a deeper understanding of the variation of GH activities in insect populations, using as models the disease vectors Rhodnius prolixus (Hemiptera: Triatominae and Lutzomyia longipalpis (Diptera: Phlebotominae. Here we standardized continuous assays using 4-methylumbelliferyl derived substrates for the detection of α-Glucosidase, β-Glucosidase, α-Mannosidase, N-acetyl-hexosaminidase, β-Galactosidase, and α-Fucosidase in the midgut of R. prolixus and L. longipalpis with results similar to the traditional discontinuous protocol. The continuous assays allowed us to measure GH activities using minimal sample amounts with a higher number of measurements, resulting in data that are more reliable and less time and reagent consumption. The continuous assay also allows the high-throughput screening of GH activities in small insect samples, which would be not applicable to the previous discontinuous protocol. We applied continuous GH measurements to 90 individual samples of R. prolixus

  5. Standardization of a Continuous Assay for Glycosidases and Its Use for Screening Insect Gut Samples at Individual and Populational Levels.

    Science.gov (United States)

    Profeta, Gerson S; Pereira, Jessica A S; Costa, Samara G; Azambuja, Patricia; Garcia, Eloi S; Moraes, Caroline da Silva; Genta, Fernando A

    2017-01-01

    Glycoside Hydrolases (GHs) are enzymes able to recognize and cleave glycosidic bonds. Insect GHs play decisive roles in digestion, in plant-herbivore, and host-pathogen interactions. GH activity is normally measured by the detection of a release from the substrate of products as sugars units, colored, or fluorescent groups. In most cases, the conditions for product release and detection differ, resulting in discontinuous assays. The current protocols result in using large amounts of reaction mixtures for the obtainment of time points in each experimental replica. These procedures restrain the analysis of biological materials with limited amounts of protein and, in the case of studies regarding small insects, implies in the pooling of samples from several individuals. In this respect, most studies do not assess the variability of GH activities across the population of individuals from the same species. The aim of this work is to approach this technical problem and have a deeper understanding of the variation of GH activities in insect populations, using as models the disease vectors Rhodnius prolixus (Hemiptera: Triatominae) and Lutzomyia longipalpis (Diptera: Phlebotominae). Here we standardized continuous assays using 4-methylumbelliferyl derived substrates for the detection of α-Glucosidase, β-Glucosidase, α-Mannosidase, N-acetyl-hexosaminidase, β-Galactosidase, and α-Fucosidase in the midgut of R. prolixus and L. longipalpis with results similar to the traditional discontinuous protocol. The continuous assays allowed us to measure GH activities using minimal sample amounts with a higher number of measurements, resulting in data that are more reliable and less time and reagent consumption. The continuous assay also allows the high-throughput screening of GH activities in small insect samples, which would be not applicable to the previous discontinuous protocol. We applied continuous GH measurements to 90 individual samples of R. prolixus anterior midgut

  6. Textual complexity of standard conditions used in the construction industry

    Directory of Open Access Journals (Sweden)

    Raufdeen Rameezdeen

    2013-03-01

    Full Text Available Clearly written communication aids the understanding of construction contracts, resulting in less disputation. Past research, using opinion surveys rather than objective criteria, shows that construction contracts lack clarity and standard forms have become complex over time. The study outlined in this paper uses three objective measures of clarity developed by linguists to establish the readability of construction contracts. In addition, thirty industry professionals participated in a Cloze Test which measured the level of comprehension of clauses concerning disputes. The study verifies that contract conditions are very difficult to read, with college level reading skills needed to comprehend half of the clauses. However, the hypothesis that standard forms have become complex over time was not supported by the study. The study establishes a linear relationship between readability and comprehension, proving the hypothesis that improved readability increases the comprehension of a contract clause.

  7. Textual complexity of standard conditions used in the construction industry

    Directory of Open Access Journals (Sweden)

    Raufdeen Rameezdeen

    2013-03-01

    Full Text Available Clearly written communication aids the understanding of construction contracts, resulting in less disputation. Past research, using opinion surveys rather than objective criteria, shows that construction contracts lack clarity and standard forms have become complex over time. The study outlined in this paper uses three objective measures of clarity developed by linguists to establish the readability of construction contracts. In addition, thirty industry professionals participated in a Cloze Test which measured the level of comprehension of clauses concerning disputes. The study verifies that contract conditions are very difficult to read, with college level reading skills needed to comprehend half of the clauses. However, the hypothesis that standard forms have become complex over time was not supported by the study. The study establishes a linear relationship between readability and comprehension, proving the hypothesis that improved readability increases the comprehension of a contract clause. 

  8. Establishment and intra-/inter-laboratory validation of a standard protocol of reactive oxygen species assay for chemical photosafety evaluation.

    Science.gov (United States)

    Onoue, Satomi; Hosoi, Kazuhiro; Wakuri, Shinobu; Iwase, Yumiko; Yamamoto, Toshinobu; Matsuoka, Naoko; Nakamura, Kazuichi; Toda, Tsuguto; Takagi, Hironori; Osaki, Naoto; Matsumoto, Yasuhiro; Kawakami, Satoru; Seto, Yoshiki; Kato, Masashi; Yamada, Shizuo; Ohno, Yasuo; Kojima, Hajime

    2013-11-01

    A reactive oxygen species (ROS) assay was previously developed for photosafety evaluation of pharmaceuticals, and the present multi-center study aimed to establish and validate a standard protocol for ROS assay. In three participating laboratories, two standards and 42 coded chemicals, including 23 phototoxins and 19 nonphototoxic drugs/chemicals, were assessed by the ROS assay according to the standardized protocol. Most phototoxins tended to generate singlet oxygen and/or superoxide under UV-vis exposure, but nonphototoxic chemicals were less photoreactive. In the ROS assay on quinine (200 µm), a typical phototoxic drug, the intra- and inter-day precisions (coefficient of variation; CV) were found to be 1.5-7.4% and 1.7-9.3%, respectively. The inter-laboratory CV for quinine averaged 15.4% for singlet oxygen and 17.0% for superoxide. The ROS assay on 42 coded chemicals (200 µm) provided no false negative predictions upon previously defined criteria as compared with the in vitro/in vivo phototoxicity, although several false positives appeared. Outcomes from the validation study were indicative of satisfactory transferability, intra- and inter-laboratory variability, and predictive capacity of the ROS assay. Copyright © 2012 John Wiley & Sons, Ltd.

  9. Evaluation the urban atmospheric conditions in different cities using comet and micronuclei assay in Tradescantia pallida.

    Science.gov (United States)

    Sposito, Juliana Caroline Vivian; Crispim, Bruno do Amaral; Romãn, Amanda Izadora; Mussury, Rosilda Mara; Pereira, Joelson Gonçalves; Seno, Leonardo Oliveira; Grisolia, Alexeia Barufatti

    2017-05-01

    In the present study, genotoxicity and mutagenicity were investigated in Tradescantia pallida exposed to vehicular traffic at different sites in a high-altitude tropical climate. During March, May, July, September, and November 2014, a comet assay and micronucleus bioassays were conducted on young inflorescences and leaves of T. pallida collected from twelve towns in the southern region of Mato Grosso do Sul with different amounts of vehicular traffic. Weather parameters (temperature, relative humidity and rainfall) were measured and vehicles were counted to determine traffic levels in each town. A higher frequency of genotoxic and mutagenic damage was observed in the municipality of Dourados. The highest frequency of genetic damage was observed in September and November according to both assays. Relative humidity and rainfall were inversely proportional to the frequency of genetic damage in T. pallida during the collection period. Based on these results, we conclude that the bioassays are efficient for assessing the effects of vehicular traffic in these towns with respect to weather conditions over time. These bioassays can be applied to identify risk areas, which are determined by climatic conditions and air pollutants released. Copyright © 2017 Elsevier Ltd. All rights reserved.

  10. Assessing leukocyte-endothelial interactions under flow conditions in an ex vivo autoperfused microflow chamber assay.

    Science.gov (United States)

    Mulki, Lama; Sweigard, J Harry; Connor, Kip M

    2014-12-30

    Leukocyte-endothelial interactions are early and critical events in acute and chronic inflammation and can, when dysregulated, mediate tissue injury leading to permanent pathological damage. Existing conventional assays allow the analysis of leukocyte adhesion molecules only after the extraction of leukocytes from the blood. This requires the blood to undergo several steps before peripheral blood leukocytes (PBLs) can be ready for analysis, which in turn can stimulate PBLs influencing the research findings. The autoperfused micro flow chamber assay, however, allows scientists to study early leukocytes functional dysregulation using the systemic flow of a live mouse while having the freedom of manipulating a coated chamber. Through a disease model, the functional expression of leukocyte adhesion molecules can be assessed and quantified in a micro-glass chamber coated with immobilized endothelial adhesion molecules ex vivo. In this model, the blood flows between the right common carotid artery and left external jugular vein of a live mouse under anesthesia, allowing the interaction of native PBLs in the chamber. Real-time experimental analysis is achieved with the assistance of an intravital microscope as well as a Harvard Apparatus pressure device. The application of a flow regulator at the input point of the glass chamber allows comparable physiological flow conditions amongst the experiments. Leukocyte rolling velocity is the main outcome and is measured using the National Institutes of Health open-access software ImageJ. In summary, the autoperfused micro flow chamber assay provides an optimal physiological environment to study leukocytes endothelial interaction and allows researchers to draw accurate conclusions when studying inflammation.

  11. 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

  12. An International Collaboration To Standardize HIV-2 Viral Load Assays: Results from the 2009 ACHIEV2E Quality Control Study▿

    OpenAIRE

    Damond, F.; Benard, A.; Balotta, Claudia; Böni, Jürg; Cotten, Matthew; Duque, Vitor; Ferns, Bridget; Garson, Jeremy; Gomes, Perpetua; Gonçalves, Fátima; Gottlieb, Geoffrey; Kupfer, Bernd; Ruelle, Jean; Rodes, Berta; Soriano, Vicente

    2011-01-01

    Accurate HIV-2 plasma viral load quantification is crucial for adequate HIV-2 patient management and for the proper conduct of clinical trials and international cohort collaborations. This study compared the homogeneity of HIV-2 RNA quantification when using HIV-2 assays from ACHIEV2E study sites and either in-house PCR calibration standards or common viral load standards supplied to all collaborators. Each of the 12 participating laboratories quantified blinded HIV-2 samples, using its own H...

  13. Are Fish and Standardized FETAX Assays Protective Enough for Amphibians? A Case Study on Xenopus laevis Larvae Assay with Biologically Active Substances Present in Livestock Wastes

    Directory of Open Access Journals (Sweden)

    Federica Martini

    2012-01-01

    Full Text Available Biologically active substances could reach the aquatic compartment when livestock wastes are considered for recycling. Recently, the standardized FETAX assay has been questioned, and some researchers have considered that the risk assessment performed on fish could not be protective enough to cover amphibians. In the present study a Xenopus laevis acute assay was developed in order to compare the sensitivity of larvae relative to fish or FETAX assays; veterinary medicines (ivermectin, oxytetracycline, tetracycline, sulfamethoxazole, and trimethoprim and essential metals (zinc, copper, manganese, and selenium that may be found in livestock wastes were used for the larvae exposure. Lethal (LC50 and sublethal effects were estimated. Available data in both, fish and FETAX studies, were in general more protective than values found out in the current study, but not in all cases. Moreover, the presence of nonlethal effects, caused by ivermectin, zinc, and copper, suggested that several physiological mechanisms could be affected. Thus, this kind of effects should be deeply investigated. The results obtained in the present study could expand the information about micropollutants from livestock wastes on amphibians.

  14. High accuracy/high precision determination of 235U in nondestructive assay standards by gamma-ray spectroscopy

    International Nuclear Information System (INIS)

    Greenberg, R.R.; Carpenter, B.S.

    1984-01-01

    High precision gamma spectrometry measurements have been made on five sets of five uranium isotope abundance reference materials for nondestructive assay (NDA). These sets are intended for international safeguards use as primary reference materials for the determination of the 235 U abundance in homogeneous uranium bulk material by gamma spectrometry. The measurements are to determine the counting rate uniformity of the 235 U 185.7 keV gamma as well as the 235 U isotope abundance for each sample. Since the samples are packaged such that the U 3 O 8 is infinitely thick for the 185.7 keV gamma, the measured counting rate is not dependent on the material density. In addition, the activity observed by the detector is colimated to simulate calibration conditions used to measure bulk material in the field. The results of this study indicate that accuracy of 235 U determination via gamma spectrometry, in the range of few hundredths of a percent (2sigma), is achievable. The main requirement for achieving this level of accuracy is a set of standards whose 235 U isotope abundance are known to within 0.01% (2sigma)

  15. Importance of 241 Am Determination in the Characterization of PuO2 Standards for Calorimetric Assay.

    Energy Technology Data Exchange (ETDEWEB)

    Sampson, Thomas E.

    2005-01-01

    Plutonium dioxide (PuO{sub 2}) standards are often used both as heat standards and isotopic standards for calorimetric assay. Calorimetric assay is the combination of the power in watts measured in a calorimeter with the effective specific power (P{sub eff}) in watts/g Pu, determined either by nondestructive gamma-ray assay or by destructive mass spectrometry, to yield the total elemental plutonium mass in the sample. To use a PuO{sub 2} sample as a heat standard for calorimetry, one must determine both the plutonium mass and P{sub eff} with very small uncertainties and then calculate the sample watts from the known plutonium mass, specific powers, and isotopic composition. Well-characterized PuO{sub 2} standards have plutonium mass values determined by analytical chemistry with a precision and accuracy on the order of 0.1%-0.2% relative to the total mass of the sample. Mass spectrometry, typically used to determine the isotopic fractions of plutonium standards, is very accurate and precise for the major isotopes but is somewhat less precise for low-abundance isotopes. The characterization of the {sup 241}Am/Pu ratio in the standard is also of great importance because {sup 241}Am can contribute significantly to P{sub eff} and to the heat output of the standard. The determination of the {sup 241}Am/Pu ratio in a plutonium-bearing sample is a process that is less standardized than mass spectrometry. There are no certified reference materials (CRMs) traceable to the national measurement system for {sup 241}Am in plutonium, and routine analytical {sup 241}Am/Pu ratio measurements often exhibit uncertainties of several percent relative to the total plutonium or greater.

  16. In vitro effects of piracetam on the radiosensitivity of hypoxic cells (adaptation of MTT assay to hypoxic conditions)

    International Nuclear Information System (INIS)

    Gheuens, E.E.O.; Bruijn, E.A. de; Van der Heyden, S.; Van Oosterom, A.T.; Lagarde, P.; Pooter, C.M.J. de; Chomy, F.

    1995-01-01

    This paper describes the adaptation of the MTT assay to hypoxic conditions in order to test the in vitro effect of piracetam on hypoxic cells and particularly on the radiosensitivity of hypoxic cells since this drug has shown clinical effect on acute and chronic hypoxia. The V79 cell line was selected by reference to preliminary hypoxic experiments using clonogenic assay and euoxic experiments using clonogenic and MTT assays. Cell growth and survival in our hypoxic conditions were assessed using MTT assay with an enclosure and special 48-well plates both made of glass. Growth curves on glass plates after 1-hour exposure to nitrogen versus air were comparable, so there is no bias effect due to gas composition. Survival curves using MTT versus reference clonogenic assay were comparable after radiation exposure in eu- and hypoxic conditions, and confirm the validity of our original technique for creating hypoxia. The Oxygen Enhancement Ratio was of about 3 for 1-hour hypoxic exposure. Piracetam gave no cytotoxic effect up to 10 mM of piracetam. Growth curves after continuous drug exposure and 1-hour euoxic versus hypoxic exposure gave no cytotoxic effect up to 10 mM of piracetam. Survival curves after continuous drug exposure to 10 mM of piracetam gave no significant effect on the radiosensitivity of hypoxic V79 cells using MTT or clonogenic assay. (author). 32 refs., 6 figs

  17. Partnership on Rotational ViscoElastic Test Standardization (PROVETS): evidence-based guidelines on rotational viscoelastic assays in veterinary medicine.

    Science.gov (United States)

    Goggs, Robert; Brainard, Benjamin; de Laforcade, Armelle M; Flatland, Bente; Hanel, Rita; McMichael, Maureen; Wiinberg, Bo

    2014-01-01

    To systematically examine the evidence relating to the performance of rotational viscoelastic testing in companion animals, to develop assay guidelines, and to identify knowledge gaps. Multiple questions were considered within 5 parent domains, specifically system comparability, sample handling, assay activation and test protocol, definitions and data reporting, and nonstandard assays. Standardized, systematic evaluation of the literature was performed. Relevant articles were categorized according to level of evidence and assessed for quality. Consensus was developed regarding conclusions for application of concepts to clinical practice. Academic and referral veterinary medical centers. Databases searched included Medline, Commonwealth Agricultural Bureaux abstracts, and Google Scholar. Worksheets were prepared evaluating 28 questions across the 5 domains and generating 84 assay guidelines. Evidence-based guidelines for the performance of thromboelastography in companion animals were generated through this process. Some of these guidelines are well supported while others will benefit from additional evidence. Many knowledge gaps were identified and future work should be directed to address these gaps and to objectively evaluate the impact of these guidelines on assay comparability within and between centers. © Veterinary Emergency and Critical Care Society 2014.

  18. Kininogen Cleavage Assay: Diagnostic Assistance for Kinin-Mediated Angioedema Conditions

    Science.gov (United States)

    Defendi, Federica; Charignon, Delphine; Ghannam, Arije; Habib, Mohammed; Drouet, Christian

    2016-01-01

    Background Angioedema without wheals (AE) is a symptom characterised by localised episodes of oedema presumably caused by kinin release from kininogen cleavage. It can result from a hereditary deficiency in C1 Inhibitor (C1Inh), but it can present with normal level of C1Inh. These forms are typically difficult to diagnose although enhanced kinin production is suspected or demonstrated in some cases. Objectives We wanted to investigate bradykinin overproduction in all AE condition with normal C1Inh, excluding cases with enhanced kinin catabolism, and to propose this parameter as a disease biomarker. Methods We retrospectively investigated high molecular weight kininogen (HK) cleavage pattern, using gel electrophoresis and immunorevelation. Plasma samples were drawn using the same standardised procedure from blood donors or AE patients with normal C1Inh conditions, normal kinin catabolism, and without prophylaxis. Results Circulating native HK plasma concentrations were similar in the healthy men (interquartile range: 98–175μg/mL, n = 51) and in healthy women (90–176μg/mL, n = 74), while HK cleavage was lower (p14.4% HK cleavage for men; 33.0% HK cleavage for women, with >98% specificity achieved for all parameters. In plasma from patients undergoing recovery two months after oestrogen/progestin combination withdrawal (n = 13) or two weeks after AE attack (n = 2), HK cleavage was not fully restored, suggesting its use as a post-attack assay. Conclusion As a diagnostic tool, HK cleavage can offer physicians supportive arguments for kinin production in suspected AE cases and improve patient follow-up in clinical trials or prophylactic management. PMID:27685806

  19. Kininogen Cleavage Assay: Diagnostic Assistance for Kinin-Mediated Angioedema Conditions.

    Directory of Open Access Journals (Sweden)

    Rémi Baroso

    Full Text Available Angioedema without wheals (AE is a symptom characterised by localised episodes of oedema presumably caused by kinin release from kininogen cleavage. It can result from a hereditary deficiency in C1 Inhibitor (C1Inh, but it can present with normal level of C1Inh. These forms are typically difficult to diagnose although enhanced kinin production is suspected or demonstrated in some cases.We wanted to investigate bradykinin overproduction in all AE condition with normal C1Inh, excluding cases with enhanced kinin catabolism, and to propose this parameter as a disease biomarker.We retrospectively investigated high molecular weight kininogen (HK cleavage pattern, using gel electrophoresis and immunorevelation. Plasma samples were drawn using the same standardised procedure from blood donors or AE patients with normal C1Inh conditions, normal kinin catabolism, and without prophylaxis.Circulating native HK plasma concentrations were similar in the healthy men (interquartile range: 98-175μg/mL, n = 51 and in healthy women (90-176μg/mL, n = 74, while HK cleavage was lower (p14.4% HK cleavage for men; 33.0% HK cleavage for women, with >98% specificity achieved for all parameters. In plasma from patients undergoing recovery two months after oestrogen/progestin combination withdrawal (n = 13 or two weeks after AE attack (n = 2, HK cleavage was not fully restored, suggesting its use as a post-attack assay.As a diagnostic tool, HK cleavage can offer physicians supportive arguments for kinin production in suspected AE cases and improve patient follow-up in clinical trials or prophylactic management.

  20. Toward an International Standard for PCR-Based Detection of Food-Borne Thermotolerant Campylobacters: Assay Development and Analytical Validation

    OpenAIRE

    Lübeck, P. S.; Wolffs, P.; On, S. L. W.; Ahrens, P.; Rådström, P.; Hoorfar, J.

    2003-01-01

    As part of a European research project (FOOD-PCR), we developed a standardized and robust PCR detection assay specific for the three most frequently reported food-borne pathogenic Campylobacter species, C. jejuni, C. coli, and C. lari. Fifteen published and unpublished PCR primers targeting the 16S rRNA gene were tested in all possible pairwise combinations, as well as two published primers targeting the 23S rRNA gene. A panel of 150 strains including target an...

  1. Standardized quantitative RT-PCR assays for quantitation of yellow fever and chimeric yellow fever-dengue vaccines.

    Science.gov (United States)

    Mantel, N; Aguirre, M; Gulia, S; Girerd-Chambaz, Y; Colombani, S; Moste, C; Barban, V

    2008-07-01

    Yellow fever-dengue chimeras (CYDs) are being developed currently as live tetravalent dengue vaccine candidates. Specific quantitative assays are needed to evaluate the viral load of each serotype in vaccine batches and biological samples. A quantitative real-time RT-PCR (qRT-PCR) system was developed comprising five one-step qRT-PCRs targeting the E/NS1 junction of each chimera, or the NS5 gene in the yellow fever backbone. Each assay was standardized using in vitro transcribed RNA qualified according to its size and purity, and precisely quantified. A non RNA-extracted virus sample was introduced as external quality control (EQC), as well as 2 extraction controls consisting of 2 doses, 40 and 4,000 GEQ (genomic equivalents), of this EQC extracted in parallel to the samples. Between 6 and 10 GEQ/reaction were reproducibly measured with all assays and similar titers were obtained with the two methods when chimeric virus samples were quantified with the E/NS1- or the NS5-specific assays. Reproducibility of RNA extraction was ensured by automation of the process (yield>or=50%), and infectious virus was isolated in >or=80% of PCR-positive sera from immune monkeys.

  2. Development of a standardized and safe airborne antibacterial assay, and its evaluation on antibacterial biomimetic model surfaces.

    Directory of Open Access Journals (Sweden)

    Ali Al-Ahmad

    Full Text Available Bacterial infection of biomaterials is a major concern in medicine, and different kinds of antimicrobial biomaterial have been developed to deal with this problem. To test the antimicrobial performance of these biomaterials, the airborne bacterial assay is used, which involves the formation of biohazardous bacterial aerosols. We here describe a new experimental set-up which allows safe handling of such pathogenic aerosols, and standardizes critical parameters of this otherwise intractable and strongly user-dependent assay. With this new method, reproducible, thorough antimicrobial data (number of colony forming units and live-dead-stain was obtained. Poly(oxonorbornene-based Synthetic Mimics of Antimicrobial Peptides (SMAMPs were used as antimicrobial test samples. The assay was able to differentiate even between subtle sample differences, such as different sample thicknesses. With this new set-up, the airborne bacterial assay was thus established as a useful, reliable, and realistic experimental method to simulate the contamination of biomaterials with bacteria, for example in an intraoperative setting.

  3. Bias in segmented gamma scans arising from size differences between calibration standards and assay samples

    International Nuclear Information System (INIS)

    Sampson, T.E.

    1991-01-01

    Recent advances in segmented gamma scanning have emphasized software corrections for gamma-ray self-adsorption in particulates or lumps of special nuclear material in the sample. another feature of this software is an attenuation correction factor formalism that explicitly accounts for differences in sample container size and composition between the calibration standards and the individual items being measured. Software without this container-size correction produces biases when the unknowns are not packaged in the same containers as the calibration standards. This new software allows the use of different size and composition containers for standards and unknowns, as enormous savings considering the expense of multiple calibration standard sets otherwise needed. This paper presents calculations of the bias resulting from not using this new formalism. These calculations may be used to estimate bias corrections for segmented gamma scanners that do not incorporate these advanced concepts

  4. Standard guide for the selection, training and qualification of nondestructive assay (NDA) personnel

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2004-01-01

    1.1 This guide contains good practices for the selection, training, qualification, and professional development of personnel performing analysis, calibration, physical measurements, or data review using nondestructive assay equipment, methods, results, or techniques. The guide also covers NDA personnel involved with NDA equipment setup, selection, diagnosis, troubleshooting, or repair. Selection, training, and qualification programs based on this guide are intended to provide assurance that NDA personnel are qualified to perform their jobs competently. This guide presents a series of options but does not recommend a specific course of action.

  5. Conformal Extensions of the Standard Model with Veltman Conditions

    DEFF Research Database (Denmark)

    Antipin, Oleg; Mojaza, Matin; Sannino, Francesco

    2014-01-01

    the Higgs is predicted to have the experimental value of the mass equal to 126 GeV. This model also predicts the existence of one more standard model singlet scalar boson with a mass of 541 GeV and the Higgs self-coupling to emerge radiatively. We study several other PNC examples that generally predict...... a somewhat smaller mass of the Higgs to the perturbative order we have investigated them. Our results can be a useful guide when building extensions of the standard model featuring fundamental scalars....

  6. Textual complexity of standard conditions used in the construction industry

    OpenAIRE

    Raufdeen Rameezdeen; Anushi Rodrigo

    2013-01-01

    Clearly written communication aids the understanding of construction contracts, resulting in less disputation. Past research, using opinion surveys rather than objective criteria, shows that construction contracts lack clarity and standard forms have become complex over time. The study outlined in this paper uses three objective measures of clarity developed by linguists to establish the readability of construction contracts. In addition, thirty industry professionals participated in a Cloze ...

  7. A new internal standard for HPLC assay of conjugated linoleic acid in animal tissues and milk

    Czech Academy of Sciences Publication Activity Database

    Czauderna, M.; Kowalczyk, J.; Marounek, Milan; Michalski, J. P.; Rozbicka-Wieczorek, A. J.; Krajewska, K. A.

    2011-01-01

    Roč. 56, č. 1 (2011), s. 23-29 ISSN 1212-1819 Institutional research plan: CEZ:AV0Z50450515 Keywords : sorbic acid * internal standard * CLA isomers Subject RIV: GH - Livestock Nutrition Impact factor: 1.079, year: 2011

  8. Standard test method for plutonium assay by plutonium (III) diode array spectrophotometry

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2002-01-01

    1.1 This test method describes the determination of total plutonium as plutonium(III) in nitrate and chloride solutions. The technique is applicable to solutions of plutonium dioxide powders and pellets (Test Methods C 697), nuclear grade mixed oxides (Test Methods C 698), plutonium metal (Test Methods C 758), and plutonium nitrate solutions (Test Methods C 759). Solid samples are dissolved using the appropriate dissolution techniques described in Practice C 1168. The use of this technique for other plutonium-bearing materials has been reported (1-5), but final determination of applicability must be made by the user. The applicable concentration range for plutonium sample solutions is 10–200 g Pu/L. 1.2 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard. 1.3 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropria...

  9. Bias in segmented gamma scans arising from size differences between calibration standards and assay samples

    International Nuclear Information System (INIS)

    Sampson, T.E.

    1991-01-01

    Recent advances in segmented gamma scanning have emphasized software corrections for gamma-ray self-absorption in particulates or lumps of special nuclear material in the sample. Another feature of this software is an attenuation correction factor formalism that explicitly accounts for differences in sample container size and composition between the calibration standards and the individual items being measured. Software without this container-size correction produces biases when the unknowns are not packaged in the same containers as the calibration standards. This new software allows the use of different size and composition containers for standards and unknowns, an enormous savings considering the expense of multiple calibration standard sets otherwise needed. This report presents calculations of the bias resulting from not using this new formalism. The calculations may be used to estimate bias corrections for segmented gamma scanners that do not incorporate these advanced concepts. This paper describes this attenuation-correction-factor formalism in more detail and illustrates the magnitude of the biases that may arise if it is not used. 5 refs., 7 figs

  10. Towards an international standard for PCR-based detection of food-borne thermotolerant Campylobacters: assay development and analytical validation.

    OpenAIRE

    Lübeck, P S; Wolffs, Petra; On, S L; Ahrens, P; Rådström, Peter; Hoorfar, J

    2003-01-01

    As part of a European research project (FOOD-PCR), we developed a standardized and robust PCR detection assay specific for the three most frequently reported food-borne pathogenic Campylobacter species, C. jejuni, C. coli, and C. lari. Fifteen published and unpublished PCR primers targeting the 16S rRNA gene were tested in all possible pairwise combinations, as well as two published primers targeting the 23S rRNA gene. A panel of 150 strains including target and nontarget strains was used in ...

  11. Standardization of incubation conditions for hemolysis testing of biomaterials

    NARCIS (Netherlands)

    Henkelman, Sandra; Rakhorst, Gerhard; Blanton, John; van Oeveren, Willem

    2009-01-01

    Hemolysis testing is the most common method to determine the hemocompatibility properties of biomaterials. There is however no consensus on the procedures of hemolysis testing due to insufficient comparative studies on the quality of the red blood cells used and the experimental conditions of

  12. 40 CFR 147.2925 - Standard permit conditions.

    Science.gov (United States)

    2010-07-01

    ... measurement; (ii) Individual(s) who preformed the measurements; (iii) Date(s) analyses were performed; (iv) Individual(s) who performed the analyses; (v) Analytical techniques or methods used, including quality... conditions. Proper operation and maintenance also includes adequate operator staffing and training, adequate...

  13. Heating, Air-Conditioning, and Refrigeration Technician. National Skill Standards.

    Science.gov (United States)

    Vocational Technical Education Consortium of States, Decatur, GA.

    This guide contains information on the knowledge and skills identified by industry as essential to the job performance of heating, air-conditioning, and refrigeration technicians. It is intended to assist training providers in public and private institutions, as well as in industry, to develop and implement training that will provide workers with…

  14. Development and Evaluation of a Duplex Real-Time PCR Assay With a Novel Internal Standard for Precise Quantification of Plasma DNA.

    Science.gov (United States)

    Chen, Dan; Pan, Shiyang; Xie, Erfu; Gao, Li; Xu, Huaguo; Xia, Wenying; Xu, Ting; Huang, Peijun

    2017-01-01

    Circulating levels of cell-free DNA increase in many pathologic conditions. However, notable discrepancies in the quantitative analysis of cell-free DNA from a large number of laboratories have become a considerable pitfall, hampering its clinical application. We designed a novel recombinant DNA fragment that could be applied as an internal standard in a newly developed and validated duplex real-time PCR assay for the quantitative analysis of total cell-free plasma DNA, which was tested in 5,442 healthy adults and 200 trauma patients. Compared with two traditional methods, this novel assay showed a lower detection limit of 0.1 ng/mL, lower intra- and inter-assay CVs, and higher accuracy in the recovery test. The median plasma DNA concentration of healthy males (20.3 ng/mL, n=3,092) was significantly higher than that of healthy females (16.1 ng/mL, n=2,350) (Mann-Whitney two-sample rank sum test, PDNA concentration were 0-45.8 ng/mL and 0-52.5 ng/mL for healthy females and males, respectively. The plasma DNA concentrations of the majority of trauma patients (96%) were higher than the upper normal cutoff values and were closely related to the corresponding injury severity scores (R²=0.916, PDNA, showing promising application in clinical diagnosis.

  15. Comparison of mRNA splicing assay protocols across multiple laboratories: recommendations for best practice in standardized clinical testing.

    Science.gov (United States)

    Whiley, Phillip J; de la Hoya, Miguel; Thomassen, Mads; Becker, Alexandra; Brandão, Rita; Pedersen, Inge Sokilde; Montagna, Marco; Menéndez, Mireia; Quiles, Francisco; Gutiérrez-Enríquez, Sara; De Leeneer, Kim; Tenés, Anna; Montalban, Gemma; Tserpelis, Demis; Yoshimatsu, Toshio; Tirapo, Carole; Raponi, Michela; Caldes, Trinidad; Blanco, Ana; Santamariña, Marta; Guidugli, Lucia; de Garibay, Gorka Ruiz; Wong, Ming; Tancredi, Mariella; Fachal, Laura; Ding, Yuan Chun; Kruse, Torben; Lattimore, Vanessa; Kwong, Ava; Chan, Tsun Leung; Colombo, Mara; De Vecchi, Giovanni; Caligo, Maria; Baralle, Diana; Lázaro, Conxi; Couch, Fergus; Radice, Paolo; Southey, Melissa C; Neuhausen, Susan; Houdayer, Claude; Fackenthal, Jim; Hansen, Thomas Van Overeem; Vega, Ana; Diez, Orland; Blok, Rien; Claes, Kathleen; Wappenschmidt, Barbara; Walker, Logan; Spurdle, Amanda B; Brown, Melissa A

    2014-02-01

    Accurate evaluation of unclassified sequence variants in cancer predisposition genes is essential for clinical management and depends on a multifactorial analysis of clinical, genetic, pathologic, and bioinformatic variables and assays of transcript length and abundance. The integrity of assay data in turn relies on appropriate assay design, interpretation, and reporting. We conducted a multicenter investigation to compare mRNA splicing assay protocols used by members of the ENIGMA (Evidence-Based Network for the Interpretation of Germline Mutant Alleles) consortium. We compared similarities and differences in results derived from analysis of a panel of breast cancer 1, early onset (BRCA1) and breast cancer 2, early onset (BRCA2) gene variants known to alter splicing (BRCA1: c.135-1G>T, c.591C>T, c.594-2A>C, c.671-2A>G, and c.5467+5G>C and BRCA2: c.426-12_8delGTTTT, c.7988A>T, c.8632+1G>A, and c.9501+3A>T). Differences in protocols were then assessed to determine which elements were critical in reliable assay design. PCR primer design strategies, PCR conditions, and product detection methods, combined with a prior knowledge of expected alternative transcripts, were the key factors for accurate splicing assay results. For example, because of the position of primers and PCR extension times, several isoforms associated with BRCA1, c.594-2A>C and c.671-2A>G, were not detected by many sites. Variation was most evident for the detection of low-abundance transcripts (e.g., BRCA2 c.8632+1G>A Δ19,20 and BRCA1 c.135-1G>T Δ5q and Δ3). Detection of low-abundance transcripts was sometimes addressed by using more analytically sensitive detection methods (e.g., BRCA2 c.426-12_8delGTTTT ins18bp). We provide recommendations for best practice and raise key issues to consider when designing mRNA assays for evaluation of unclassified sequence variants.

  16. Smelling Pseudomonas aeruginosa infections using a whole-cell biosensor - An alternative for the gold-standard culturing assay.

    Science.gov (United States)

    Kviatkovski, Igor; Shushan, Sagit; Oron, Yahav; Frumin, Idan; Amir, Daniel; Secundo, Lavi; Livne, Eitan; Weissbrod, Aharon; Sobel, Noam; Helman, Yael

    2018-02-10

    Improved easy-to-use diagnostic tools for infections are in strong demand worldwide. Yet, despite dramatic advances in diagnostic technologies, the gold-standard remains culturing. Here we offer an alternative tool demonstrating that a bacterial biosensor can efficiently detect Pseudomonas aeruginosa infections in patients suffering from otitis externa. Detection was based on specific binding between the biosensor and 2-aminoacetophenone (2-AA), a volatile produced by P. aeruginosa in high amounts. We collected pus samples from ears of 26 subjects exhibiting symptoms of otitis externa. Detection of P. aeruginosa using the biosensor was compared to detection using gold-standard culturing assay and to gas-chromatograph-mass-spectrometry (GC-MS) analyses of 2-AA. The biosensor strain test matched the culture assay in 24 samples (92%) and the GC-MS analyses in 25 samples (96%). With this result in hand, we designed a device containing a whole-cell luminescent biosensor combined with a photo-multiplier tube. This device allowed detection of 2-AA at levels as low as 2 nmol, on par with detection level of GC-MS. The results of the described study demonstrate that the volatile 2-AA serves as an effective biomarker for P. aeruginosa in ear infections, and that activation of the biosensor strain by 2-AA provides a unique opportunity to design an easy-to-use device that can specifically detect P. aeruginosa infections. Copyright © 2017 Elsevier B.V. All rights reserved.

  17. Assessing Immunity to Rubella Virus: a Plea for Standardization of IgG (Immuno)assays

    Science.gov (United States)

    Bouthry, Elise; Huzly, Daniela; Ogee-Nwankwo, Adaeze; Hao, LiJuan; Adebayo, Adebola; Icenogle, Joseph; Sarasini, Antonella; Revello, Maria Grazia; Grangeot-Keros, Liliane

    2016-01-01

    Immunity to rubella virus (RV) is commonly determined by measuring specific immunoglobulin G (RV IgG). However, RV IgG results and their interpretation may vary, depending on the immunoassay, even though most commercial immunoassays (CIAs) have been calibrated against an international standard and results are reported in international units per milliliter. A panel of 322 sera collected from pregnant women that tested negative or equivocal for RV IgG in a prior test (routine screening) was selected. This panel was tested with two reference tests, immunoblotting (IB) and neutralization (Nt), and with 8 CIAs widely used in Europe. IB and Nt gave concordant results on 267/322 (82.9%) sera. Of these, 85 (26.4%) sera were negative and 182 (56.5%) sera were positive for both tests. All 85 IB/Nt-negative samples were classified as negative with all CIAs. Of the 182 IB/Nt-positive samples, 25.3 to 61.5% were classified as equivocal and 6 to 64.8% were classified as positive with the CIAs. Wide variations in titers in international units per milliliter were observed. In our series, more than half of the women considered susceptible to RV based on CIA results tested positive for RV antibodies by IB/Nt. Our data suggest that (i) sensitivity of CIAs could be increased by considering equivocal results as positive and (ii) the definition of immunity to RV as the 10-IU/ml usual cutoff as well as the use of quantitative results for clinical decisions may warrant reconsideration. A better standardization of CIAs for RV IgG determination is needed. PMID:27147722

  18. Standardization and performance evaluation of mononuclear cell cytokine secretion assays in a multicenter study

    Directory of Open Access Journals (Sweden)

    Grindle Kristine

    2006-12-01

    Full Text Available Abstract Background Cryopreservation of peripheral blood mononuclear cells has been used to preserve and standardize immunologic measurements for multicenter studies, however, effects of cryopreservation on cytokine responses are incompletely understood. In designing immunologic studies for a new multicenter birth cohort study of childhood asthma, we performed a series of experiments to determine the effects of two different methods of cryopreservation on the cytokine responses of cord and peripheral blood mononuclear cells. Results Paired samples of PBMC were processed freshly, or after cryopreservation in a Nalgene container (NC or a controlled-rate freezer (CRF. Although there were some differences between the methods, cryopreservation inhibited PHA-induced IL-10 secretion and Der f 1-induced IL-2 secretion, and augmented PHA-induced IL-2 secretion and spontaneous secretion of TNF-α. In separate experiments, NC cryopreservation inhibited secretion of several cytokines (IL-13, IL-10, IFN-γ, TNF-α by PHA-stimulated cord blood mononuclear cells. With the exception of PHA-induced IL-13, results from fresh and cryopreserved cord blood samples were not significantly correlated. Finally, in reproducibility studies involving processing of identical cell samples in up to 4 separate laboratories, variances in cytokine responses of fresh cells stimulated at separate sites did not exceed those in cryopreserved cells stimulated at a central site. Conclusion Collectively, these studies indicate that cryopreservation can affect mononuclear cell cytokine response profiles, and that IL-10 secretion and antigen-induced responses may be especially vulnerable. These studies also demonstrate that mononuclear cell responses can be standardized for performance in a small number of laboratories for multicenter studies, and underscore the importance of measuring reproducibility and of testing whether cryopreservation techniques alter specific immunologic outcomes.

  19. Surveillance evaluation of the standardization of assay values for serum total 25-hydroxyvitamin D concentration in Japan.

    Science.gov (United States)

    Ihara, Hiroshi; Kiuchi, Sachiko; Ishige, Takayuki; Nishimura, Motoi; Matsushita, Kazuyuki; Satoh, Mamoru; Nomura, Fumio; Yamashita, Mine; Kitajima, Isao; Tsugawa, Naoko; Okano, Toshio; Hirota, Kouichi; Miura, Masakazu; Totani, Masayuki; Hashizume, Naotaka

    2018-01-01

    Background To assess the vitamin D nutritional status, serum total 25-hydroxyvitamin D (25(OH)D) concentration is measured. We used six automated 25(OH)D immunoassays (AIAs) available in Japan and certified by the Vitamin D Standardization Program (VDSP) at the U.S. Center for Disease Control and Prevention to assess the concordance of the assay results. Methods Serum total 25(OH)D concentrations in SRM 972a and 20 serum samples from patients were determined using three liquid chromatography-tandem mass spectrometry (LC-MS/MS) and six AIAs (pilot study), and an additional 110 serum samples were assessed by the six AIAs (surveillance study). The assay bias from the results of LC-MS/MS by Chiba University or consensus values (i.e. average of six AIAs) was estimated using the procedure described in CLSI document EP09-A3. Results LC-MS/MS at Chiba University could completely separate 25(OH)D2, 25(OH)D3 and 3-epi-25(OH)D3, and the observed values including total 25(OH)D in SRM 972a were all within ±1·SD of the assigned values. All AIAs produced results greater than ±3·SD. In the pilot study, four of the six AIAs had an average percentage bias, as estimated by confidence interval (CI), larger than ±5% (acceptance criterion in CLSI); the bias converged from -6.5% to 3.2% after adjustment by LC-MS/MS. In the surveillance study, 25(OH)D concentrations in AIAs all adjusted to LC-MS/MS converged within ±5% from consensus values. However, some AIAs showed negative or positive bias from the consensus values. Conclusions Current AIAs in Japan continue to lack standardization. Manufacturers should implement quality assurance strategies so that their values more closely align to those of standard reference material 972a.

  20. Analyzing pepsin degradation assay conditions used for allergenicity assessments to ensure that pepsin susceptible and pepsin resistant dietary proteins are distinguishable.

    Directory of Open Access Journals (Sweden)

    Rong Wang

    Full Text Available The susceptibility of a dietary protein to proteolytic degradation by digestive enzymes, such as gastric pepsin, provides information on the likelihood of systemic exposure to a structurally intact and biologically active macromolecule, thus informing on the safety of proteins for human and animal consumption. Therefore, the purpose of standardized in vitro degradation studies that are performed during protein safety assessments is to distinguish whether proteins of interest are susceptible or resistant to pepsin degradation via a study design that enables study-to-study comparison. Attempting to assess pepsin degradation under a wide-range of possible physiological conditions poses a problem because of the lack of robust and consistent data collected under a large-range of sub-optimal conditions, which undermines the needs to harmonize in vitro degradation conditions. This report systematically compares the effects of pH, incubation time, and pepsin-to-substrate protein ratio on the relative degradation of five dietary proteins: three pepsin susceptible proteins [ribulose 1,5-bisphosphate carboxylase-oxygenase (Rubisco, horseradish peroxidase (HRP, hemoglobin (Hb], and two pepsin resistant proteins [lipid transfer protein (LTP and soybean trypsin inhibitor (STI]. The results indicate that proteins susceptible to pepsin degradation are readily distinguishable from pepsin-resistant proteins when the reaction conditions are within the well-characterized optima for pepsin. The current standardized in vitro pepsin resistant assay with low pH and high pepsin-to-substrate ratio fits this purpose. Using non-optimal pH and/or pepsin-to-substrate protein ratios resulted in susceptible proteins no longer being reliably degraded by this stomach enzyme, which compromises the ability of this in vitro assay to distinguish between resistant and susceptible proteins and, therefore, no longer providing useful data to an overall weight-of-evidence approach to

  1. Developmental Screening Using the Ages and Stages Questionnaire: Standardized versus Real-World Conditions

    Science.gov (United States)

    San Antonio, Marianne C.; Fenick, Ada M.; Shabanova, Veronika; Leventhal, John M.; Weitzman, Carol C.

    2014-01-01

    Developmental screens are often used in nonstandardized conditions, such as pediatric waiting rooms, despite validation under standardized conditions. We examined the reproducibility of the Ages and Stages Questionnaire (ASQ), a developmental screening instrument commonly used in pediatric practices, under standardized versus nonstandardized…

  2. 24 CFR 886.113 - Physical condition standard; physical inspection requirements.

    Science.gov (United States)

    2010-04-01

    ... 24 Housing and Urban Development 4 2010-04-01 2010-04-01 false Physical condition standard; physical inspection requirements. 886.113 Section 886.113 Housing and Urban Development Regulations... § 886.113 Physical condition standard; physical inspection requirements. (a) General. Housing used in...

  3. Matrix and position correction of shuffler assays by application of the alternating conditional expectation algorithm to shuffler data

    International Nuclear Information System (INIS)

    Pickrell, M.M.; Rinard, P.M.

    1992-01-01

    The 252 Cf shuffler assays fissile uranium and plutonium using active neutron interrogation and then counting the induced delayed neutrons. Using the shuffler, we conducted over 1700 assays of 55-gal. drums with 28 different matrices and several different fissionable materials. We measured the drums to dispose the matrix and position effects on 252 Cf shuffler assays. We used several neutron flux monitors during irradiation and kept statistics on the count rates of individual detector banks. The intent of these measurements was to gauge the effect of the matrix independently from the uranium assay. Although shufflers have previously been equipped neutron monitors, the functional relationship between the flux monitor sepals and the matrix-induced perturbation has been unknown. There are several flux monitors so the problem is multivariate, and the response is complicated. Conventional regression techniques cannot address complicated multivariate problems unless the underlying functional form and approximate parameter values are known in advance. Neither was available in this case. To address this problem, we used a new technique called alternating conditional expectations (ACE), which requires neither the functional relationship nor the initial parameters. The ACE algorithm develops the functional form and performs a numerical regression from only the empirical data. We applied the ACE algorithm to the shuffler-assay and flux-monitor data and developed an analytic function for the matrix correction. This function was optimized using conventional multivariate techniques. We were able to reduce the matrix-induced-bias error for homogeneous samples to 12.7%. The bias error for inhomogeneous samples was reduced to 13.5%. These results used only a few adjustable parameters compared to the number of available data points; the data were not ''over fit,'' but rather the results are general and robust

  4. Assay of antioxidant potential of two Aspergillus isolates by different methods under various physio-chemical conditions.

    Science.gov (United States)

    Arora, Daljit Singh; Chandra, Priyanka

    2010-07-01

    The objective of this work was to screen fungi isolated from soil of different areas of Punjab, India for antioxidant activity by dot blot assay and around 45% of fungal isolates demonstrated antioxidant potential. Two selected strains of Aspergillus spp (Aspergillus PR78 and Aspergillus PR66) showing quantitatively best antioxidant activity by DPPH assay were further tested for their reducing power, ferrous ion and nitric oxide ion scavenging activity, FRAP assay and total phenolic content. Different physio-chemical parameters were optimized for enhancement of the activity. This revealed stationary culture grown for 10 days at 25 (o)C at pH 7 to be the best for antioxidant activity. Sucrose in the medium as carbon source resulted in highest antioxidant activity. Sodium nitrate, yeast extract, and peptone were good sources of nitrogen but sodium nitrate was the best among these. The extraction of the broth culture filtrates with different solvents revealed ethyl acetate extract to possess the best antioxidant activity. The activity as expressed by ethyl acetate extract of Aspergillus PR78 was equally effective as that of commonly used antioxidant standard, ascorbic acid.

  5. Assay of antioxidant potential of two Aspergillus isolates by different methods under various physio-chemical conditions

    Directory of Open Access Journals (Sweden)

    Daljit Singh Arora

    2010-10-01

    Full Text Available The objective of this work was to screen fungi isolated from soil of different areas of Punjab, India for antioxidant activity by dot blot assay and around 45% of fungal isolates demonstrated antioxidant potential. Two selected strains of Aspergillus spp (Aspergillus PR78 and Aspergillus PR66 showing quantitatively best antioxidant activity by DPPH assay were further tested for their reducing power, ferrous ion and nitric oxide ion scavenging activity, FRAP assay and total phenolic content. Different physio-chemical parameters were optimized for enhancement of the activity. This revealed stationary culture grown for 10 days at 25ºC at pH 7 to be the best for antioxidant activity. Sucrose in the medium as carbon source resulted in highest antioxidant activity. Sodium nitrate, yeast extract, and peptone were good sources of nitrogen but sodium nitrate was the best among these. The extraction of the broth culture filtrates with different solvents revealed ethyl acetate extract to possess the best antioxidant activity. The activity as expressed by ethyl acetate extract of Aspergillus PR78 was equally effective as that of commonly used antioxidant standard, ascorbic acid.

  6. Influence of oxygen on asexual blood cycle and susceptibility of Plasmodium falciparum to chloroquine: requirement of a standardized in vitro assay

    Directory of Open Access Journals (Sweden)

    Minodier Philippe

    2007-04-01

    Full Text Available Abstract Objective The main objective of this study was to assess the influence of gas mixtures on in vitro Plasmodium falciparum growth and 50% inhibitory concentration (IC50 for chloroquine. Methods The study was performed between February 2004 and December 2005. 136 Plasmodium falciparum isolates were used to evaluate gas mixtures effect on IC50 for chloroquine by isotopic microtest. The oxygen effect on asexual blood cycle of 3D7 and W2 clones was determined by thin blood smears examination and tritiated hypoxanthine uptake. Results From 5% O2 to 21% O2 conditions, no parasiticide effect of O2 concentration was observed in vitro on the clones 3D7 and W2. A parasitostatic effect was observed during the exposure of mature trophozoïtes and schizonts at 21% O2 with an increase in the length of schizogony. The chloroquine IC50 at 10% O2 were significantly higher than those at 21% O2, means of 173.5 nM and 121.5 nM respectively (p in vitro resistant to chloroquine (IC50 > 100 nM at 10% O2, 17 were sensitive to chloroquine (IC50 2. Conclusion Based on these results, laboratories should use the same gas mixture to realize isotopic microtest. Further studies on comparison of isotopic and non-isotopic assays are needed to establish a standardized in vitro assay protocol to survey malaria drug resistance.

  7. Reproducibility of microbial mutagenicity assays. I. Tests with Salmonella typhimurium and Escherichia coli using a standardized protocol

    International Nuclear Information System (INIS)

    Dunkel, V.C.; Zeiger, E.; Brusick, D.; McCoy, E.; McGregor, D.; Mortelmans, K.; Rosenkranz, H.S.; Simmon, V.F.

    1984-01-01

    The Salmonella/microsome test developed by Ames and his coworkers has been widely used in the evaluation of chemicals for genotoxic potential. Although the value of this assay is well recognized, there have been no comprehensive studies on the interlaboratory reproducibility of the method using a standardized protocol. A program was therefore initiated to compare the results obtained in four laboratories from testing a series of coded mutagens and nonmutagens using a standardized protocol. Additional objectives of this study were to compare male Fisher 344 rat, B6C3F1 mouse, and Syrian hamster liver S-9 preparations for the activation of chemicals; to compare Aroclor 1254-induced liver S-9 from all three species with the corresponding non-induced liver S-9's; and to compare the response of Escherichia coli WP-2 uvrA with the Salmonella typhimurium tester strains recommended by Ames. Since a primary use of in vitro microbial mutagenesis tests is the identification of potential carcinogens by their mutagenicity, the authors decided to compare the animal species and strains used by the National Cancer Institute/National Toxicology Program (NCI/NTP) for animal carcinogenicity studies

  8. The history and development of FETAX (ASTM standard guide, E-1439 on conducting the frog embryo teratogenesis Assay-Xenopus)

    Science.gov (United States)

    Dumont, J.N.; Bantle, J.A.; Linder, G.; ,

    2003-01-01

    The energy crisis of the 1970's and 1980's prompted the search for alternative sources of fuel. With development of alternate sources of energy, concerns for biological resources potentially adversely impacted by these alternative technologies also heightened. For example, few biological tests were available at the time to study toxic effects of effluents on surface waters likely to serve as receiving streams for energy-production facilities; hence, we began to use Xenopus laevis embryos as test organisms to examine potential toxic effects associated with these effluents upon entering aquatic systems. As studies focused on potential adverse effects on aquatic systems continued, a test procedure was developed that led to the initial standardization of FETAX. Other .than a limited number of aquatic toxicity tests that used fathead minnows and cold-water fishes such as rainbow trout, X. laevis represented the only other aquatic vertebrate test system readily available to evaluate complex effluents. With numerous laboratories collaborating, the test with X. laevis was refined, improved, and developed as ASTM E-1439, Standard Guide for the Conducting Frog Embryo Teratogenesis Assay-Xenopus (FETAX). Collabrative work in the 1990s yielded procedural enhancements, for example, development of standard test solutions and exposure methods to handle volatile organics and hydrophobic compounds. As part of the ASTM process, a collaborative interlaboratory study was performed to determine the repeatability and reliability of FETAX. Parallel to these efforts, methods were also developed to test sediments and soils, and in situ test methods were developed to address "lab-to-field extrapolation errors" that could influence the method's use in ecological risk assessments. Additionally, a metabolic activation system composed of rat liver microsomes was developed which made FETAX more relevant to mammalian studies.

  9. Relationship between in vitro assays and standardized ileal amino acid digestibility of animal protein meals in broilers.

    Science.gov (United States)

    Rochell, S J; Kuhlers, D L; Dozier, W A

    2013-01-01

    Two identical trials were conducted to determine the relationship of a novel digestive enzyme assay, Poultry Complete IDEA (PC IDEA), and the pepsin digestibility assay with standardized ileal amino acid digestibility (SIAAD) of 20 animal protein meals (APM) fed to broilers from 25 to 30 d of age. Animal protein meals included 10 meat and bone meals (MBM) consisting of bovine, porcine, or mixed bovine and porcine raw materials (BP), and 10 animal protein blends containing animal proteins from various species. Treatments consisted of 20 semi-purified diets containing 1 APM as the sole source of dietary amino acids (AA), and 1 N-free diet to determine endogenous ileal AA flow. With the exception of the N-free diet, diets were formulated to contain 20% CP. In each trial, 756 Ross × Ross 708 male broilers were housed in battery cages and randomly assigned to 21 dietary treatments on d 25 (12 birds per cage; 3 replicate cages), and ileal digesta were collected on d 30 for determination of SIAAD. Pepsin digestibility and PC IDEA were determined for APM samples from each experimental diet (3 replicates per trial; 6 total replicates). Pepsin digestibility and PC IDEA were both correlated (P digestibility on SIAAD resulted in the following equations: % Lys SIAAD = [-9.65 + (0.38 × % PC IDEA predicted Lys digestibility) + (0.69 × % pepsin digestibility)], % Met SIAAD = [-35.95 + (0.62 × % PC IDEA predicted Met digestibility) + (0.75 × % pepsin digestibility)], % Thr SIAAD = [-77.5 + (0.39 × % PC IDEA predicted Thr digestibility) + (1.37 × % pepsin digestibility)]. Values of R(2) were 0.46, 0.47, and 0.55 for Lys, Met, and Thr, respectively. The relatively low R(2) values may have been due to the limited range in SIAAD observed for the 20 APM, and additional data on APM varying in SIAAD are needed.

  10. Eyeblink conditioning and novel object recognition in the rabbit: Behavioral paradigms for assaying psychiatric diseases

    Directory of Open Access Journals (Sweden)

    Craig eWeiss

    2015-10-01

    Full Text Available Analysis of data collected from behavioral paradigms has provided important information for understanding the etiology and progression of diseases that involve neural regions mediating abnormal behavior. The trace eyeblink conditioning (EBC paradigm is particularly suited to examine cerebro-cerebellar interactions since the paradigm requires the cerebellum, forebrain, and awareness of the stimulus contingencies. Impairments in acquiring EBC have been noted in several neuropsychiatric conditions including schizophrenia, Alzheimer’s disease (AD, progressive supranuclear palsy (PSP and post-traumatic stress disorder (PTSD. Although several species have been used to examine EBC, the rabbit is unique in its tolerance for restraint which facilitates imaging, its relatively large skull which facilitates chronic neuronal recordings, a genetic sequence for amyloid that is identical to humans which makes it a valuable model to study AD, and in contrast to rodents, it has a striatum that is differentiated into a caudate and a putamen which facilitates analysis of diseases involving the striatum. This review focuses on EBC during schizophrenia and AD since impairments in cerebro-cerebellar connections have been hypothesized to lead to a cognitive dysmetria. We also relate EBC to conditioned avoidance responses that are more often examined for effects of antipsychotic medications, and we propose that an analysis of novel object recognition (NOR may add to our understanding of how the underlying neural circuitry has changed during disease states. We propose that the EBC and NOR paradigms will help to determine which therapeutics are effective for treating the cognitive aspects of schizophrenia and AD, and that neuroimaging may reveal biomarkers of the diseases and help to evaluate potential therapeutics. The rabbit thus provides an important translational system for studying neural mechanisms mediating maladaptive behaviors that underlie some psychiatric

  11. Russian standards and design practice of ensuring NPP reliability under severe external loading conditions

    International Nuclear Information System (INIS)

    Birbraer, A.N.

    1993-01-01

    Russian Standards and design practice of ensuring NPP reliability under severe external loading conditions are described. The main attention is paid to the seismic design requirements. Explosions, aircraft impact, and tornado are briefly examined too (author)

  12. Evaluation of Multiplexed Foot-and-Mouth Disease Nonstructural Protein Antibody Assay Against Standardized Bovine Serum Panel

    Energy Technology Data Exchange (ETDEWEB)

    Perkins, J; Parida, S; Clavijo, A

    2007-05-14

    Liquid array technology has previously been used to show proof-of-principle of a multiplexed non structural protein serological assay to differentiate foot-and-mouth infected and vaccinated animals. The current multiplexed assay consists of synthetically produced peptide signatures 3A, 3B and 3D and recombinant protein signature 3ABC in combination with four controls. To determine diagnostic specificity of each signature in the multiplex, the assay was evaluated against a naive population (n = 104) and a vaccinated population (n = 94). Subsequently, the multiplexed assay was assessed using a panel of bovine sera generated by the World Reference Laboratory for foot-and-mouth disease in Pirbright, UK. This sera panel has been used to assess the performance of other singleplex ELISA-based non-structural protein antibody assays. The 3ABC signature in the multiplexed assay showed comparative performance to a commercially available non-structural protein 3ABC ELISA (Cedi test{reg_sign}) and additional information pertaining to the relative diagnostic sensitivity of each signature in the multiplex is acquired in one experiment. The encouraging results of the evaluation of the multiplexed assay against a panel of diagnostically relevant samples promotes further assay development and optimization to generate an assay for routine use in foot-and-mouth disease surveillance.

  13. Cryptic fitness advantage: diploids invade haploid populations despite lacking any apparent advantage as measured by standard fitness assays.

    Directory of Open Access Journals (Sweden)

    Aleeza C Gerstein

    Full Text Available Ploidy varies tremendously within and between species, yet the factors that influence when or why ploidy variants are adaptive remains poorly understood. Our previous work found that diploid individuals repeatedly arose within ten replicate haploid populations of Saccharomyces cerevisiae, and in each case we witnessed diploid takeover within ~1800 asexual generations of batch culture evolution in the lab. The character that allowed diploids to rise in frequency within haploid populations remains unknown. Here we present a number of experiments conducted with the goal to determine what this trait (or traits might have been. Experiments were conducted both by sampling a small number of colonies from the stocks frozen every two weeks (~ 93 generations during the original experiment, as well through sampling a larger number of colonies at the two time points where polymorphism for ploidy was most prevalent. Surprisingly, none of our fitness component measures (lag phase, growth rate, biomass production indicated an advantage to diploidy. Similarly, competition assays against a common competitor and direct competition between haploid and diploid colonies isolated from the same time point failed to indicate a diploid advantage. Furthermore, we uncovered a tremendous amount of trait variation among colonies of the same ploidy level. Only late-appearing diploids showed a competitive advantage over haploids, indicating that the fitness advantage that allowed eventual takeover was not diploidy per se but an attribute of a subset of diploid lineages. Nevertheless, the initial rise in diploids to intermediate frequency cannot be explained by any of the fitness measures used; we suggest that the resolution to this mystery is negative frequency-dependent selection, which is ignored in the standard fitness measures used.

  14. Recent advances in the characterization of HIV-1 neutralization assays for standardized evaluation of the antibody response to infection and vaccination.

    Science.gov (United States)

    Polonis, Victoria R; Brown, Bruce K; Rosa Borges, Andrew; Zolla-Pazner, Susan; Dimitrov, Dimiter S; Zhang, Mei-Yun; Barnett, Susan W; Ruprecht, Ruth M; Scarlatti, Gabriella; Fenyö, Eva-Maria; Montefiori, David C; McCutchan, Francine E; Michael, Nelson L

    2008-06-05

    In AIDS vaccine development the pendulum has swung towards a renewed emphasis on the potential role for neutralizing antibodies in a successful global vaccine. It is recognized that vaccine-induced antibody performance, as assessed in the available neutralization assays, may well serve as a "gatekeeper" for HIV-1 subunit vaccine prioritization and advancement. As a result, development of a standardized platform for reproducible measurement of neutralizing antibodies has received considerable attention. Here we review current advancements in our knowledge of the performance of different types of antibodies in a traditional primary cell neutralization assay and the newer, more standardized TZM-bl reporter cell line assay. In light of recently revealed differences (see accompanying article) in the results obtained in these two neutralization formats, parallel evaluation with both platforms should be contemplated as an interim solution until a better understanding of immune correlates of protection is achieved.

  15. A scalable assessment of Plasmodium falciparum transmission in the standard membrane-feeding assay, using transgenic parasites expressing green fluorescent protein-luciferase

    NARCIS (Netherlands)

    Stone, W.J.R.; Churcher, T.S.; Graumans, W.; Gemert, G.J.A. van; Vos, M.W.; Lanke, K.H.W.; Vegte-Bolmer, M.G. van de; Siebelink-Stoter, R.; Dechering, K.J.; Vaughan, A.M.; Camargo, N.; Kappe, S.H.; Sauerwein, R.W.; Bousema, T.

    2014-01-01

    BACKGROUND: The development of drugs and vaccines to reduce malaria transmission is an important part of eradication plans. The transmission-reducing activity (TRA) of these agents is currently determined in the standard membrane-feeding assay (SMFA), based on subjective microscopy-based readouts

  16. Cholylglycine hydrolase and 7alpha-dehydroxylase optimum assay conditions in vitro and caecal enzyme activities ex vivo.

    Science.gov (United States)

    Thomas, L A; King, A; French, G L; Murphy, G M; Dowling, R H

    1997-12-10

    Increasing evidence implicates deoxycholic acid (DCA) in the pathogenesis of cholesterol-rich gallbladder stones. However, relatively little is known about the activities of the two intestinal bacterial enzymes (cholylglycine hydrolase and cholic acid 7alpha-dehydroxylase) responsible for the deconjugation and subsequent dehydroxylation of conjugated cholic acid (CA), to form DCA. We, therefore, established optimal reaction conditions for measuring the activities of these two enzymes in vitro, and applied these conditions to the determination of the enzymes in caecal aspirates from six subjects undergoing clinically-indicated colonoscopy. With respect to cholylglycine hydrolase activity: zero order kinetics were found over 20 min at 37 degrees C (pH optimum 4.0), with Km and Vmax values of 1.66 mmol/l and 0.90 mmol CA min(-1) mg prot(-1), respectively. For cholic acid 7alpha-dehydroxylation: zero order kinetics were found over 7.5 min at 37 degrees C, under anaerobic conditions (pH optimum 8.0), with Km and Vmax values of 5.23 x 10(-8) mol/l and 1.88 x 10(-7) mol DCA min(-1) mg prot(-1), respectively. Applying these reaction conditions to the caecal aspirates, endogenous cholylglycine hydrolase activities ranged from 0.49 to 2.43 units (mg protein[-1] min[-1]) and CA 7alpha-dehydroxylase activities from 1.75 to 5.82 x 10(-7) units (mg protein[-1] min[-1]). This study is unique in assaying quantitatively both the deconjugation and dehydroxylation enzyme activities in human caecal samples--an essential first step to further studies of intestinal bacterial enzymes in the pathogenesis of cholesterol gallstone disease.

  17. An international collaboration to standardize HIV-2 viral load assays: results from the 2009 ACHI(E)V(2E) quality control study.

    Science.gov (United States)

    Damond, F; Benard, A; Balotta, Claudia; Böni, Jürg; Cotten, Matthew; Duque, Vitor; Ferns, Bridget; Garson, Jeremy; Gomes, Perpetua; Gonçalves, Fátima; Gottlieb, Geoffrey; Kupfer, Bernd; Ruelle, Jean; Rodes, Berta; Soriano, Vicente; Wainberg, Mark; Taieb, Audrey; Matheron, Sophie; Chene, Genevieve; Brun-Vezinet, Francoise

    2011-10-01

    Accurate HIV-2 plasma viral load quantification is crucial for adequate HIV-2 patient management and for the proper conduct of clinical trials and international cohort collaborations. This study compared the homogeneity of HIV-2 RNA quantification when using HIV-2 assays from ACHI(E)V(2E) study sites and either in-house PCR calibration standards or common viral load standards supplied to all collaborators. Each of the 12 participating laboratories quantified blinded HIV-2 samples, using its own HIV-2 viral load assay and standard as well as centrally validated and distributed common HIV-2 group A and B standards (http://www.hiv.lanl.gov/content/sequence/HelpDocs/subtypes-more.html). Aliquots of HIV-2 group A and B strains, each at 2 theoretical concentrations (2.7 and 3.7 log(10) copies/ml), were tested. Intralaboratory, interlaboratory, and overall variances of quantification results obtained with both standards were compared using F tests. For HIV-2 group A quantifications, overall and interlaboratory and/or intralaboratory variances were significantly lower when using the common standard than when using in-house standards at the concentration levels of 2.7 log(10) copies/ml and 3.7 log(10) copies/ml, respectively. For HIV-2 group B, a high heterogeneity was observed and the variances did not differ according to the type of standard used. In this international collaboration, the use of a common standard improved the homogeneity of HIV-2 group A RNA quantification only. The diversity of HIV-2 group B, particularly in PCR primer-binding regions, may explain the heterogeneity in quantification of this strain. Development of a validated HIV-2 viral load assay that accurately quantifies distinct circulating strains is needed.

  18. Differences in serum thyroglobulin measurements by 3 commercial immunoradiometric assay kits and laboratory standardization using Certified Reference Material 457 (CRM-457).

    Science.gov (United States)

    Lee, Ji In; Kim, Ji Young; Choi, Joon Young; Kim, Hee Kyung; Jang, Hye Won; Hur, Kyu Yeon; Kim, Jae Hyeon; Kim, Kwang-Won; Chung, Jae Hoon; Kim, Sun Wook

    2010-09-01

    Serum thyroglobulin (Tg) is essential in the follow-up of patients with differentiated thyroid carcinoma (DTC). However, interchangeability and standardization between Tg assays have not yet been achieved, even with the development of an international Tg standard (Certified Reference Material 457 [CRM-457]). Serum Tg from 30 DTC patients and serially diluted CRM-457 were measured using 3 different immunoradiometric assays (IRMA-1, IRMA-2, IRMA-3). The intraclass correlation coefficient (ICC) method was used to describe the concordance of each IRMA to CRM-457. The serum Tg measured by 3 different IRMAs correlated well (r > .85, p CRM-457, showed the best ICC (p(1) = .98) for the CRM-457. Hospitals caring for patients with DTC should either set their own cutoffs for IRMAs for Tg based on their patient pools, or adopt IRMAs standardized to CRM-457 and calibrate their laboratory using CRM-457.

  19. Evaluation of real-time PCR assays and standard curve optimisation for enhanced accuracy in quantification of Campylobacter environmental water isolates.

    Science.gov (United States)

    Gosselin-Théberge, Maxime; Taboada, Eduardo; Guy, Rebecca A

    2016-10-01

    Campylobacter is a major public health and economic burden in developed and developing countries. This study evaluated published real-time PCR (qPCR) assays for detection of Campylobacter to enable selection of the best assays for quantification of C. spp. and C. jejuni in environmental water samples. A total of 9 assays were compared: three for thermotolerant C. spp. targeting the 16S rRNA and six for C. jejuni targeting different genes. These assays were tested in the wet-lab for specificity and sensitivity against a collection of 60, genetically diverse, Campylobacter isolates from environmental water. All three qPCR assays targeting C. spp. were positive when tested against the 60 isolates, whereas, assays targeting C. jejuni differed among each other in terms of specificity and sensitivity. Three C. jejuni-specific assays that demonstrated good specificity and sensitivity when tested in the wet-lab showed concordant results with in silico-predicted results obtained against a set of 211 C. jejuni and C. coli genome sequences. Two of the assays targeting C. spp. and C. jejuni were selected to compare DNA concentration estimation, using spectrophotometry and digital PCR (dPCR), in order to calibrate standard curves (SC) for greater accuracy of qPCR-based quantification. Average differences of 0.56±0.12 and 0.51±0.11 log fold copies were observed between the spectrophotometry-based SC preparation and the dPCR preparation for C. spp. and C. jejuni, respectively, demonstrating an over-estimation of Campylobacter concentration when spectrophotometry was used to calibrate the DNA SCs. Our work showed differences in quantification of aquatic environmental isolates of Campylobacter between qPCR assays and method-specific bias in SC preparation. This study provided an objective analysis of qPCR assays targeting Campylobacter in the literature and provides a framework for evaluating novel assays. Crown Copyright © 2016. Published by Elsevier B.V. All rights reserved.

  20. Responses in young Quercus petraea: coppices and standards under favourable and drought conditions

    Czech Academy of Sciences Publication Activity Database

    Stojanović, Marko; Čater, M.; Pokorný, Radek

    2016-01-01

    Roč. 76, jan (2016), s. 127-136 ISSN 1641-1307 R&D Projects: GA MŠk(CZ) EE2.3.20.0267 Institutional support: RVO:67179843 Keywords : coppice * standards * comparison * photosynthetic response * quantum yield * light conditions * drought response Subject RIV: EF - Botanics Impact factor: 0.776, year: 2016

  1. Validation of a single nucleotide polymorphism (SNP) typing assay with 49 SNPs for forensic genetic testing in a laboratory accredited according to the ISO 17025 standard

    DEFF Research Database (Denmark)

    Børsting, Claus; Rockenbauer, Eszter; Morling, Niels

    2009-01-01

    cases and 33 twin cases were typed at least twice for the 49 SNPs. All electropherograms were analysed independently by two expert analysts prior to approval. Based on these results, detailed guidelines for analysis of the SBE products were developed. With these guidelines, the peak height ratio...... of a heterozygous allele call or the signal to noise ratio of a homozygous allele call is compared with previously obtained ratios. A laboratory protocol for analysis of SBE products was developed where allele calls with unusual ratios were highlighted to facilitate the analysis of difficult allele calls......A multiplex assay with 49 autosomal single nucleotide polymorphisms (SNPs) developed for human identification was validated for forensic genetic casework and accredited according to the ISO 17025 standard. The multiplex assay was based on the SNPforID 52plex SNP assay [J.J. Sanchez, C. Phillips, C...

  2. Comparative Study of Kaposi's Sarcoma-Associated Herpesvirus Serological Assays Using Clinically and Serologically Defined Reference Standards and Latent Class Analysis▿

    Science.gov (United States)

    Nascimento, Maria Claudia; de Souza, Vanda Akico; Sumita, Laura Masami; Freire, Wilton; Munoz, Fernando; Kim, Joseph; Pannuti, Claudio S.; Mayaud, Philippe

    2007-01-01

    Accurate determination of infection with Kaposi's sarcoma-associated herpesvirus (KSHV) has been hindered by the lack of a “gold standard” for comparison of serological assays used to estimate KSHV prevalence in serosurveys conducted in different settings. We have evaluated the performance of five in-house (developed at University College London [UCL], United Kingdom, and at the virology laboratory of the Instituto de Medicine Tropical [IMT] in Sao Paulo, Brazil) and two commercial (ABI and DIAVIR) serological assays to detect antibodies to latency-associated nuclear antigen (LANA) and to lytic KSHV antigens. We used a variety of serum samples assembled to represent populations likely to be at high, intermediate, and low risk of KSHV infection in Brazil. Composite reference standard panels were prepared based on clinical and serological parameters, against which assay performances were assessed using conventional Bayesian statistics and latent class analysis (LCA). Against the clinical reference standard, in-house immunofluorescence assays to detect anti-LANA antibodies (IFA-LANA) produced at UCL and IMT had similar performances, with sensitivities of 61% (95% confidence interval [CI], 48% to 74%) and 72% (95% CI, 58% to 83%) and specificities of 99% (95% CI, 94% to 100%) and 100% (95% CI, 96% to 100%), respectively, and only the IMT IFA-LANA was included in LCA, together with the IMT IFA-lytic and four enzyme-linked immunosorbent assays (ELISAs). The LCA indicated that the IMT whole-virus ELISA performed best (sensitivity, 87% [95% CI, 81% to 91%]; and specificity, 100% [95% CI, 98% to 100%]), confirming the results obtained with the conventional statistical approach. Commercially available ELISA-based tests yielded the lowest specificities using a spectrum of serum samples. The evaluation of KSHV serological assays is warranted before planning serosurveys in various settings. PMID:17182752

  3. Standard Evaluation Procedures (SEPs) and Data Entry Spreadsheet Templates (DESTs) for Endocrine Disruptor Screening Program (EDSP) Tier 1 Assays

    Science.gov (United States)

    This page provides information and access to Standard Evaluation Procedures (SEPs) and Data Entry Spreadsheet Templates (DESTs) developed by EPA's Office of Chemical Safety and Pollution Prevention (OCSPP).

  4. Size- and coating-dependent cytotoxicity and genotoxicity of silver nanoparticles evaluated using in vitro standard assays.

    Science.gov (United States)

    Guo, Xiaoqing; Li, Yan; Yan, Jian; Ingle, Taylor; Jones, Margie Yvonne; Mei, Nan; Boudreau, Mary D; Cunningham, Candice K; Abbas, Mazhar; Paredes, Angel M; Zhou, Tong; Moore, Martha M; Howard, Paul C; Chen, Tao

    2016-11-01

    The physicochemical characteristics of silver nanoparticles (AgNPs) may greatly alter their toxicological potential. To explore the effects of size and coating on the cytotoxicity and genotoxicity of AgNPs, six different types of AgNPs, having three different sizes and two different coatings, were investigated using the Ames test, mouse lymphoma assay (MLA) and in vitro micronucleus assay. The genotoxicities of silver acetate and silver nitrate were evaluated to compare the genotoxicity of nanosilver to that of ionic silver. The Ames test produced inconclusive results for all types of the silver materials due to the high toxicity of silver to the test bacteria and the lack of entry of the nanoparticles into the cells. Treatment of L5718Y cells with AgNPs and ionic silver resulted in concentration-dependent cytotoxicity, mutagenicity in the Tk gene and the induction of micronuclei from exposure to nearly every type of the silver materials. Treatment of TK6 cells with these silver materials also resulted in concentration-dependent cytotoxicity and significantly increased micronucleus frequency. With both the MLA and micronucleus assays, the smaller the AgNPs, the greater the cytotoxicity and genotoxicity. The coatings had less effect on the relative genotoxicity of AgNPs than the particle size. Loss of heterozygosity analysis of the induced Tk mutants indicated that the types of mutations induced by AgNPs were different from those of ionic silver. These results suggest that AgNPs induce cytotoxicity and genotoxicity in a size- and coating-dependent manner. Furthermore, while the MLA and in vitro micronucleus assay (in both types of cells) are useful to quantitatively measure the genotoxic potencies of AgNPs, the Ames test cannot.

  5. Thyroglobulin assay in fluids from lymph node fine needle-aspiration washout: influence of pre-analytical conditions.

    Science.gov (United States)

    Casson, Florence Boux de; Moal, Valérie; Gauchez, Anne-Sophie; Moineau, Marie-Pierre; Sault, Corinne; Schlageter, Marie-Hélène; Massart, Catherine

    2017-04-01

    The aim of this study was to evaluate the pre-analytical factors contributing to uncertainty in thyroglobulin measurement in fluids from fine-needle aspiration (FNA) washout of cervical lymph nodes. We studied pre-analytical stability, in different conditions, of 41 samples prepared with concentrated solutions of thyroglobulin (FNA washout or certified standard) diluted in physiological saline solution or buffer containing 6% albumin. In this buffer, over time, no changes in thyroglobulin concentrations were observed in all storage conditions tested. In albumin free saline solution, thyroglobulin recovery rates depended on initial sample concentrations and on modalities of their conservation (in conventional storage tubes, recovery mean was 56% after 3 hours-storage at room temperature and 19% after 24 hours-storage for concentrations ranged from 2 to 183 μg/L; recovery was 95%, after 3 hours or 24 hours-storage at room temperature, for a concentration of 5,656 μg/L). We show here that these results are due to non-specific adsorption of thyroglobulin in storage tubes, which depends on sample protein concentrations. We also show that possible contamination of fluids from FNA washout by plasma proteins do not always adequately prevent this adsorption. In conclusion, non-specific adsorption in storage tubes strongly contributes to uncertainty in thyroglobulin measurement in physiological saline solution. It is therefore recommended, for FNA washout, to use a buffer containing proteins provided by the laboratory.

  6. Welfare and biosecurity standards for dairy cow and pig farms: Cattle and swine rearing conditions

    Directory of Open Access Journals (Sweden)

    Hristov Slavča

    2009-01-01

    Full Text Available In this paper the essential elements concerning cattle and swine rearing and growing conditions were given in order to establish welfare and biosecurity standards. These elements were formed according to detailed annual investigations on 11 cattle and 5 swine farms and include relevant spatial, microclimate and hygienic conditions. In order to establish welfare standards, certain spatial conditions have higher importance, such as correct construction and maintenance of beds, pens and yards, and type and quality of materials used to build beds and walls. It is necessary to enable movement of animals in stables and yards as basic physiological and ethologic needs, according to latest scientific data. Also, optimal temperature, relative humidity and air velocity insuring have to be considered, as well as quality ventilation in order to establish and preserve optimal microclimate conditions. Also, it must be pointed out that hygiene maintenance of stable surfaces and animal bodies on a regular bases is essential. Basic principles and criteria for welfare level assessment are given in this paper. According to results obtained in previous investigations, special attention is given to possibilities to correct rearing and growing conditions in cattle and swine farms in our country. .

  7. Planck scale boundary conditions in the standard model with singlet scalar dark matter

    Energy Technology Data Exchange (ETDEWEB)

    Haba, Naoyuki [Graduate School of Science and Engineering, Shimane University, Matsue, Shimane 690-8504 (Japan); Kaneta, Kunio [Kavli IPMU (WPI), The University of Tokyo, Kashiwa, Chiba 277-8568 (Japan); Takahashi, Ryo [Graduate School of Science and Engineering, Shimane University, Matsue, Shimane 690-8504 (Japan)

    2014-04-04

    We investigate Planck scale boundary conditions on the Higgs sector of the standard model with a gauge singlet scalar dark matter. We will find that vanishing self-coupling and Veltman condition at the Planck scale are realized with the 126 GeV Higgs mass and top pole mass, 172 GeV≲M{sub t}≲173.5 GeV, where a correct abundance of scalar dark matter is obtained with mass of 300 GeV≲m{sub S}≲1 TeV. It means that the Higgs potential is flat at the Planck scale, and this situation can not be realized in the standard model with the top pole mass.

  8. Quantum Hall resistance standard in graphene devices under relaxed experimental conditions

    Science.gov (United States)

    Schopfer, F.; Ribeiro-Palau, R.; Lafont, F.; Brun-Picard, J.; Kazazis, D.; Michon, A.; Cheynis, F.; Couturaud, O.; Consejo, C.; Jouault, B.; Poirier, W.

    Large-area and high-quality graphene devices synthesized by CVD on SiC are used to develop reliable electrical resistance standards, based on the quantum Hall effect (QHE), with state-of-the-art accuracy of 1x10-9 and under an extended range of experimental conditions of magnetic field (down to 3.5 T), temperature (up to 10 K) or current (up to 0.5 mA). These conditions are much relaxed as compared to what is required by GaAs/AlGaAs standards and will enable to broaden the use of the primary quantum electrical standards to the benefit of Science and Industry for electrical measurements. Furthermore, by comparison of these graphene devices with GaAs/AlGaAs standards, we demonstrate the universality of the QHE within an ultimate uncertainty of 8.2x10-11. This suggests the exact relation of the quantized Hall resistance with the Planck constant and the electron charge, which is crucial for the new SI to be based on fixing such fundamental constants. These results show that graphene realizes its promises and demonstrates its superiority over other materials for a demanding application. Nature Nanotech. 10, 965-971, 2015, Nature Commun. 6, 6806, 2015

  9. In vitro effects of piracetam on the radiosensitivity of hypoxic cells (adaptation of MTT assay to hypoxic conditions); Effets in vitro du piracetam sur la radiosensibilite des cellules hypoxiques (adapatation du test au MTT aux conditions d`hypoxie)

    Energy Technology Data Exchange (ETDEWEB)

    Gheuens, E.E.O.; Bruijn, E.A. de; Van der Heyden, S.; Van Oosterom, A.T. [Universitaire Instelling Antwerpen, Antwerp (Belgium); Lagarde, P. [Universitaire Instelling Antwerpen, Antwerp (Belgium)]|[Institut Bergonie, 33 - Bordeaux (France); Pooter, C.M.J. de [Universitaire Instelling Antwerpen, Antwerp (Belgium)]|[Hopital de Middelheim, Anvers (Belgium); Chomy, F. [Institut Bergonie, 33 - Bordeaux (France)

    1995-12-31

    This paper describes the adaptation of the MTT assay to hypoxic conditions in order to test the in vitro effect of piracetam on hypoxic cells and particularly on the radiosensitivity of hypoxic cells since this drug has shown clinical effect on acute and chronic hypoxia. The V79 cell line was selected by reference to preliminary hypoxic experiments using clonogenic assay and euoxic experiments using clonogenic and MTT assays. Cell growth and survival in our hypoxic conditions were assessed using MTT assay with an enclosure and special 48-well plates both made of glass. Growth curves on glass plates after 1-hour exposure to nitrogen versus air were comparable, so there is no bias effect due to gas composition. Survival curves using MTT versus reference clonogenic assay were comparable after radiation exposure in eu- and hypoxic conditions, and confirm the validity of our original technique for creating hypoxia. The Oxygen Enhancement Ratio was of about 3 for 1-hour hypoxic exposure. Piracetam gave no cytotoxic effect up to 10 mM of piracetam. Growth curves after continuous drug exposure and 1-hour euoxic versus hypoxic exposure gave no cytotoxic effect up to 10 mM of piracetam. Survival curves after continuous drug exposure to 10 mM of piracetam gave no significant effect on the radiosensitivity of hypoxic V79 cells using MTT or clonogenic assay. (author). 32 refs., 6 figs.

  10. A semi-automated luminescence based standard membrane feeding assay identifies novel small molecules that inhibit transmission of malaria parasites by mosquitoes.

    Science.gov (United States)

    Vos, Martijn W; Stone, Will J R; Koolen, Karin M; van Gemert, Geert-Jan; van Schaijk, Ben; Leroy, Didier; Sauerwein, Robert W; Bousema, Teun; Dechering, Koen J

    2015-12-21

    Current first-line treatments for uncomplicated falciparum malaria rapidly clear the asexual stages of the parasite, but do not fully prevent parasite transmission by mosquitoes. The standard membrane feeding assay (SMFA) is the biological gold standard assessment of transmission reducing activity (TRA), but its throughput is limited by the need to determine mosquito infection status by dissection and microscopy. Here we present a novel dissection-free luminescence based SMFA format using a transgenic Plasmodium falciparum reporter parasite without resistance to known antimalarials and therefore unrestricted in its utility in compound screening. Analyses of sixty-five compounds from the Medicines for Malaria Venture validation and malaria boxes identified 37 compounds with high levels of TRA (>80%); different assay modes allowed discrimination between gametocytocidal and downstream modes of action. Comparison of SMFA data to published assay formats for predicting parasite infectivity indicated that individual in vitro screens show substantial numbers of false negatives. These results highlight the importance of the SMFA in the screening pipeline for transmission reducing compounds and present a rapid and objective method. In addition we present sixteen diverse chemical scaffolds from the malaria box that may serve as a starting point for further discovery and development of malaria transmission blocking drugs.

  11. Radiometric measurements on the fabrication of non-destructive assay standards for WIPP-Performance Demonstration Program

    International Nuclear Information System (INIS)

    Wong, A.S.; Marshall, R.S.

    1997-04-01

    The Inorganic Elemental Analysis Group of LANL has prepared several different sets of working reference materials (WRMs). These WRMs are prepared by blending quantities of nuclear materials (plutonium, americium, and enriched uranium) with diatomaceous earth. The blends are encapsulated in stainless steel cylinders. These WRMs are being measured as blind controls in neutron and gamma based non-destructive assay (NDA) instruments. Radiometric measurements on the blending homogeneity and verification on a set of sixty three plutonium based WRMs are discussed in this paper

  12. Pet Food Palatability Evaluation: A Review of Standard Assay Techniques and Interpretation of Results with a Primary Focus on Limitations.

    Science.gov (United States)

    Aldrich, Gregory C; Koppel, Kadri

    2015-01-16

    The pet food industry continues to grow steadily as a result of new innovative products. Quality control and product development tests for pet foods are typically conducted through palatability testing with dogs and cats. Palatability is the measure of intake of a food that indicates acceptance or the measure of preference of one food over another. Pet food palatability is most commonly measured using a single-bowl or a two-bowl assay. While these tests answer some questions about the animals' perception of the food, there are many limitations as well. This review addresses some of these limitations and indicates opportunities for future research.

  13. Pet Food Palatability Evaluation: A Review of Standard Assay Techniques and Interpretation of Results with a Primary Focus on Limitations

    Directory of Open Access Journals (Sweden)

    Gregory C. Aldrich

    2015-01-01

    Full Text Available The pet food industry continues to grow steadily as a result of new innovative products. Quality control and product development tests for pet foods are typically conducted through palatability testing with dogs and cats. Palatability is the measure of intake of a food that indicates acceptance or the measure of preference of one food over another. Pet food palatability is most commonly measured using a single-bowl or a two-bowl assay. While these tests answer some questions about the animals’ perception of the food, there are many limitations as well. This review addresses some of these limitations and indicates opportunities for future research.

  14. Comparison of isolated cranberry (Vaccinium macrocarpon Ait.) proanthocyanidins to catechin and procyanidins A2 and B2 for use as standards in the 4-(dimethylamino)cinnamaldehyde assay.

    Science.gov (United States)

    Feliciano, Rodrigo P; Shea, Michael P; Shanmuganayagam, Dhanansayan; Krueger, Christian G; Howell, Amy B; Reed, Jess D

    2012-05-09

    The 4-(dimethylamino)cinnamaldehyde (DMAC) assay is currently used to quantify proanthocyanidin (PAC) content in cranberry products. However, this method suffers from issues of accuracy and precision in the analysis and comparison of PAC levels across a broad range of cranberry products. Current use of procyanidin A2 as a standard leads to an underestimation of PACs content in certain cranberry products, especially those containing higher molecular weight PACs. To begin to address the issue of accuracy, a method for the production of a cranberry PAC standard, derived from an extraction of cranberry (c-PAC) press cake, was developed and evaluated. Use of the c-PAC standard to quantify PAC content in cranberry samples resulted in values that were 2.2 times higher than those determined by procyanidin A2. Increased accuracy is critical for estimating PAC content in relationship to research on authenticity, efficacy, and bioactivity, especially in designing clinical trials for determination of putative health benefits.

  15. Facilitated acquisition of standard but not long delay classical eyeblink conditioning in behaviorally inhibited adolescents.

    Science.gov (United States)

    Caulfield, M D; VanMeenen, K M; Servatius, R J

    2015-02-01

    Adolescence is a key age in the development of anxiety disorders. The present study assessed the relationship between behavioral inhibition, a risk factor for anxiety typified by avoidance, and acquisition of the classically conditioned eyeblink response. 168 healthy high school students (mean age 15.7 years, 54% female) were given a battery of self-report measures including the Adult Measure of Behavioural Inhibition (AMBI). The study compared acquisition of three experimental training conditions. Two groups were given paired CS-US training: standard delay of 500-ms or long delay of 1000-ms with CS overlapping and co-terminating with a 50-ms airpuff US. A third group received unpaired training of 1000-ms CS and 50-ms airpuff US. Inhibited individuals showed greater acquisition of the conditioned eyeblink response in the 500-ms CS condition, but not in the paired 1000-ms condition. No differences in spontaneous blinks or reactivity to the stimulus were evident in the 1000-ms unpaired CS condition. Results support a relationship between associative learning and anxiety vulnerability that may be mediated by cerebellar functioning in inhibited individuals. Copyright © 2014 Elsevier B.V. All rights reserved.

  16. [The requirements of standard and conditions of interchangeability of medical articles].

    Science.gov (United States)

    Men'shikov, V V; Lukicheva, T I

    2013-11-01

    The article deals with possibility to apply specific approaches under evaluation of interchangeability of medical articles for laboratory analysis. The development of standardized analytical technologies of laboratory medicine and formulation of requirements of standards addressed to manufacturers of medical articles the clinically validated requirements are to be followed. These requirements include sensitivity and specificity of techniques, accuracy and precision of research results, stability of reagents' quality in particular conditions of their transportation and storage. The validity of requirements formulated in standards and addressed to manufacturers of medical articles can be proved using reference system, which includes master forms and standard samples, reference techniques and reference laboratories. This approach is supported by data of evaluation of testing systems for measurement of level of thyrotrophic hormone, thyroid hormones and glycated hemoglobin HB A1c. The versions of testing systems can be considered as interchangeable only in case of results corresponding to the results of reference technique and comparable with them. In case of absence of functioning reference system the possibilities of the Joined committee of traceability in laboratory medicine make it possible for manufacturers of reagent sets to apply the certified reference materials under development of manufacturing of sets for large listing of analytes.

  17. Synthesis of Ti Oxides at Reducing Conditions: Implications for Beamline Standards and Cosmochemistry

    Science.gov (United States)

    Righter, K.; Pando, K. A.; Butterworth, A. L.; Gainsforth, Z.; Jilly-Rehak, C. E.; Westphal, A. J.

    2017-01-01

    These initial experiments demonstrate the great potential for synthesizing customized compounds for use as standards, or in buffering experiments at reducing conditions. We are also investigating Cr and V oxides, as well as compounds containing these elements such as FeV2O4 and FeCr2O4. Oxygen fugacity exerts a major control on mineral major element chemistry and elemental valence of minerals in any plane-tary compositional system [1]. For Earth, Fe is multivalent ranging from nearly Fe0 at low fO2 in the deep mantle to Fe2+ to Fe3+ at high low fO2. For solar nebular and meteoritic materials fO2 ranges from near IW to 10 log fO2 units below the IW buffer [1]. Phases in CAIs, for example, contain no Fe2+, but may contain Ti4+, Ti3+, or Ti2+, and Cr3+ or Cr2+, and V3+ or V2+ [1,2,3]. De-tailed study of inclusions may reveal important differences in fO2 thus reflecting different environments in the solar nebula [4]. XANES, FEG-SEM, and TEM can reveal such variations in micro and nano samples such as Stardust and cosmic dust particles [5], but successful application to reduced conditions depends upon the availability of well characterized standards. Acquiring appropriate standards for reduced phases that contain Ti3+ or Ti2+, Cr3+ or Cr2+, and V3+ or V2+ can be a challenge. Here we report our preliminary results at synthesizing reduced Ti bearing standards, and focus on the preliminary characterization.

  18. Practical issues for testing thin film PV modules at standard test conditions.

    OpenAIRE

    Marín González, Omar; Raga Arroyo, Manuela Pilar; Alonso Garcia, M. Carmen; Muñoz-García, Miguel Angel

    2013-01-01

    Thin film photovoltaic (TF) modules have gained importance in the photovoltaic (PV) market. New PV plants increasingly use TF technologies. In order to have a reliable sample of a PV module population, a huge number of modules must be measured. There is a big variety of materials used in TF technology. Some of these modules are made of amorphous or microcrystalline silicon. Other are made of CIS or CdTe. Not all these materials respond the same under standard test conditions (STC) of power...

  19. Life quality and living standards in big cities under conditions of high-rise construction development

    Science.gov (United States)

    Avdeeva, Elena; Averina, Tatiana; Kochetova, Larisa

    2018-03-01

    Modern urbanization processes occurring on a global scale inevitably lead to an increase in population density in large cities. People assess the state of life quality and living standards of megalopolises under conditions of high-rise construction development ambiguously. Using SWOT analysis, the authors distinguished positive and negative aspects of high-rise construction, highlighted threats to its development and its opportunities. The article considers the model of development of the city's industry and infrastructure, which enables determining the optimal volume of production by sectors and branches of city economy in order to increase its innovative, production and economic potential and business activity.

  20. A comparison of titers of anti-Brucella antibodies of naturally infected and healthy vaccinated cattle by standard tube agglutination test, microtiter plate agglutination test, indirect hemagglutination assay, and indirect enzyme-linked immunosorbent assay

    Directory of Open Access Journals (Sweden)

    Anju Mohan

    2016-07-01

    Full Text Available Aim: We determined the antibody response in cattle naturally infected with brucellosis and normal healthy adult cattle vaccinated during calf hood with strain 19. Materials and Methods: The antibody titers were measured by standard tube agglutination test (STAT, microtiter plate agglutination test (MAT, indirect hemagglutination assay (IHA, and indirect enzyme-linked immunosorbent assay (iELISA as per standard protocols. Results: The mean STAT titers were 1.963±0.345 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was extremely significant (p<0.0001. The mean MAT titers were 2.244±0.727 in infected cattle and 1.200±0.155 in healthy vaccinated cattle. The difference was very significant (p<0.005. The mean IHA titers in infected cattle were 2.284±0.574, and those in healthy vaccinated cattle were 1.200±0.155. The difference was extremely significant (p=0.0002. However, the difference in mean iELISA titers of infected cattle (1.3678±0.014 and healthy vaccinated cattle (1.367±0.014 was non-significant. The infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals. However, it cannot be ascertained whether these antibodies are due to vaccine or response to infection. Since the infected animals had been vaccinated earlier, the current infection may suggest that vaccination was unable to induce protective levels of antibody. The heightened antibody response after infection may also indicate a secondary immune response to the antigens common to the vaccine strain and wild Brucella organisms. Conclusion: The brucellosis infected animals showed very high titers of agglutinating antibodies compared to the vaccinated animals.

  1. A Comparison of Three Methods for Computing Scale Score Conditional Standard Errors of Measurement. ACT Research Report Series, 2013 (7)

    Science.gov (United States)

    Woodruff, David; Traynor, Anne; Cui, Zhongmin; Fang, Yu

    2013-01-01

    Professional standards for educational testing recommend that both the overall standard error of measurement and the conditional standard error of measurement (CSEM) be computed on the score scale used to report scores to examinees. Several methods have been developed to compute scale score CSEMs. This paper compares three methods, based on…

  2. A scalable assessment of Plasmodium falciparum transmission in the standard membrane-feeding assay, using transgenic parasites expressing green fluorescent protein-luciferase.

    Science.gov (United States)

    Stone, Will J R; Churcher, Thomas S; Graumans, Wouter; van Gemert, Geert-Jan; Vos, Martijn W; Lanke, Kjerstin H W; van de Vegte-Bolmer, Marga G; Siebelink-Stoter, Rianne; Dechering, Koen J; Vaughan, Ashley M; Camargo, Nelly; Kappe, Stefan H I; Sauerwein, Robert W; Bousema, Teun

    2014-11-01

    The development of drugs and vaccines to reduce malaria transmission is an important part of eradication plans. The transmission-reducing activity (TRA) of these agents is currently determined in the standard membrane-feeding assay (SMFA), based on subjective microscopy-based readouts and with limitations in upscaling and throughput. Using a Plasmodium falciparum strain expressing the firefly luciferase protein, we present a luminescence-based approach to SMFA evaluation that eliminates the requirement for mosquito dissections in favor of a simple approach in which whole mosquitoes are homogenized and examined directly for luciferase activity. Analysis of 6860 Anopheles stephensi mosquitoes across 68 experimental feeds shows that the luminescence assay was as sensitive as microscopy for infection detection. The mean luminescence intensity of individual and pooled mosquitoes accurately quantifies mean oocyst intensity and generates comparable TRA estimates. The luminescence assay presented here could increase SMFA throughput so that 10-30 experimental feeds could be evaluated in a single 96-well plate. This new method of assessing Plasmodium infection and transmission intensity could expedite the screening of novel drug compounds, vaccine candidates, and sera from malaria-exposed individuals for TRA. Luminescence-based estimates of oocyst intensity in individual mosquitoes should be interpreted with caution. © The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  3. New normative standards of conditional reasoning and the dual-source model.

    Science.gov (United States)

    Singmann, Henrik; Klauer, Karl Christoph; Over, David

    2014-01-01

    There has been a major shift in research on human reasoning toward Bayesian and probabilistic approaches, which has been called a new paradigm. The new paradigm sees most everyday and scientific reasoning as taking place in a context of uncertainty, and inference is from uncertain beliefs and not from arbitrary assumptions. In this manuscript we present an empirical test of normative standards in the new paradigm using a novel probabilized conditional reasoning task. Our results indicated that for everyday conditional with at least a weak causal connection between antecedent and consequent only the conditional probability of the consequent given antecedent contributes unique variance to predicting the probability of conditional, but not the probability of the conjunction, nor the probability of the material conditional. Regarding normative accounts of reasoning, we found significant evidence that participants' responses were confidence preserving (i.e., p-valid in the sense of Adams, 1998) for MP inferences, but not for MT inferences. Additionally, only for MP inferences and to a lesser degree for DA inferences did the rate of responses inside the coherence intervals defined by mental probability logic (Pfeifer and Kleiter, 2005, 2010) exceed chance levels. In contrast to the normative accounts, the dual-source model (Klauer et al., 2010) is a descriptive model. It posits that participants integrate their background knowledge (i.e., the type of information primary to the normative approaches) and their subjective probability that a conclusion is seen as warranted based on its logical form. Model fits showed that the dual-source model, which employed participants' responses to a deductive task with abstract contents to estimate the form-based component, provided as good an account of the data as a model that solely used data from the probabilized conditional reasoning task.

  4. New Normative Standards of Conditional Reasoning and the Dual-Source Model

    Directory of Open Access Journals (Sweden)

    Henrik eSingmann

    2014-04-01

    Full Text Available There has been a major shift in research on human reasoning towards Bayesian and probabilistic approaches, which has been called a new paradigm. The new paradigm sees most everyday and scientific reasoning as taking place in a context of uncertainty, and inference is from uncertain beliefs and not from arbitrary assumptions. In this manuscript we present an empirical test of normative standards in the new paradigm using a novel probabilized conditional reasoning task. Our results indicated that for everyday conditional with at least a weak causal connection between antecedent and consequent only the conditional probability of the consequent given antecedent contributes unique variance to predicting the probability of conditional, but not the probability of the conjunction, nor the probability of the material conditional. Regarding normative accounts of reasoning, we found significant evidence that participants' responses were confidence preserving (i.e., p-valid in the sense of Adams, 1998 for MP inferences, but not for MT inferences. Additionally, only for MP inferences and to a lesser degree for DA inferences did the rate of responses inside the coherence intervals defined by mental probability logic (Pfeifer & Kleiter, 2005, 2010 exceed chance levels. In contrast to the normative accounts, the dual-source model (Klauer, Beller, & Hütter, 2010 is a descriptive model. It posits that participants integrate their background knowledge (i.e., the type of information primary to the normative approaches and their subjective probability that a conclusion is seen as warranted based on its logical form. Model fits showed that the dual-source model, which employed participants' responses to a deductive task with abstract contents to estimate the form-based component, provided as good an account of the data as a model that solely used data from the probabilized conditional reasoning task.

  5. Electroluminescent TCC, C3dg and fB/Bb epitope assays for profiling complement cascade activation in vitro using an activated complement serum calibration standard.

    Science.gov (United States)

    van Vuuren, B Jansen; Bergseth, G; Mollnes, T E; Shaw, A M

    2014-01-15

    Electroluminescent assays for epitopes on the complement components C3dg, terminal complement complex (TCC) and factor B/Bb (fB/Bb) have been developed with capture and detection antibodies to produce detection limits C3dg=91±9ng/mL, TCC=3±0.1ng/mL and fB=55.7±0.1ng/mL. The assay performance was assessed against a series of zymosan and heat aggregated IgG (HAIgG) in vitro activations of complement using a calibrated activated complement serum (ACS) as calibration standard. The ACS standard was stable within 20% accuracy over a 6-month period with freeze-thaw cycles as required. Differential activation of the complement cascade was observed for TCC showing a pseudo-first order formation half-life of 3.5h after activation with zymosan. The C3dg activation fragment indicates a 10% total activation for both activation agents. The kinetic-epitope analysis for fB indicates that the capture epitope is on the fB/Bb protein fragment which can then become covered by the formation of C3bBb or C3bBbP complexes during the time course of the cascade. Copyright © 2013 Elsevier B.V. All rights reserved.

  6. Standardization of the energy performance of photovoltaic modules in real operating conditions

    Directory of Open Access Journals (Sweden)

    Viganó Davide

    2014-01-01

    Full Text Available The performance of a PV module at STC [1] is a useful indicator for comparing the peak performance of different module types, but on its own is not sufficient to accurately predict how much energy a module will deliver in the field when subjected to a wide range of real operating conditions [2]. An Energy Rating approach has to be preferred for that aim. It is currently under development the standard series IEC 61853 on Energy Rating, for which only part 1 [3] has been issued. It describes methods to characterize the module performance as a function of irradiance and temperature. The reproducibility of the power matrix measurements obtained by the three different methods specified in the standard, namely: under natural sunlight using a tracking system; under natural sunlight without tracker; and a large area pulsed solar simulator of Class AAA were evaluated and discussed [4,5]. The work here presented is focused on the second method listed above, which explores the real working conditions for a PV device and therefore it represents the situation where Energy Rating procedures are expected to give the largest deviations from the STC predictions. The system for continuous monitoring of module performances, already implemented at ESTI, has been recently replaced with a new system having a number of improvements described in the following. The two system results have been compared showing a discrete compatibility. The two power matrices are then merged together using a weighted average and compared to those acquired with the other two remaining “ideal” systems. An interesting tendency seems to come up from this comparison, making the power rating under real operating conditions an essential procedure for energy rating purposes.

  7. Robust Road Condition Detection System Using In-Vehicle Standard Sensors.

    Science.gov (United States)

    Castillo Aguilar, Juan Jesús; Cabrera Carrillo, Juan Antonio; Guerra Fernández, Antonio Jesús; Carabias Acosta, Enrique

    2015-12-19

    The appearance of active safety systems, such as Anti-lock Braking System, Traction Control System, Stability Control System, etc., represents a major evolution in road safety. In the automotive sector, the term vehicle active safety systems refers to those whose goal is to help avoid a crash or to reduce the risk of having an accident. These systems safeguard us, being in continuous evolution and incorporating new capabilities continuously. In order for these systems and vehicles to work adequately, they need to know some fundamental information: the road condition on which the vehicle is circulating. This early road detection is intended to allow vehicle control systems to act faster and more suitably, thus obtaining a substantial advantage. In this work, we try to detect the road condition the vehicle is being driven on, using the standard sensors installed in commercial vehicles. Vehicle models were programmed in on-board systems to perform real-time estimations of the forces of contact between the wheel and road and the speed of the vehicle. Subsequently, a fuzzy logic block is used to obtain an index representing the road condition. Finally, an artificial neural network was used to provide the optimal slip for each surface. Simulations and experiments verified the proposed method.

  8. A Comprehensive Subcellular Proteomic Survey of Salmonella Grown under Phagosome-Mimicking versus Standard Laboratory Conditions

    Energy Technology Data Exchange (ETDEWEB)

    Brown, Roslyn N.; Sanford, James A.; Park, Jea H.; Deatherage, Brooke L.; Champion, Boyd L.; Smith, Richard D.; Heffron, Fred; Adkins, Joshua N.

    2012-06-01

    Towards developing a systems-level pathobiological understanding of Salmonella enterica, we performed a subcellular proteomic analysis of this pathogen grown under standard laboratory and infection-mimicking conditions in vitro. Analysis of proteins from cytoplasmic, inner membrane, periplasmic, and outer membrane fractions yielded coverage of over 30% of the theoretical proteome. Confident subcellular location could be assigned to over 1000 proteins, with good agreement between experimentally observed location and predicted/known protein properties. Comparison of protein location under the different environmental conditions provided insight into dynamic protein localization and possible moonlighting (multiple function) activities. Notable examples of dynamic localization were the response regulators of two-component regulatory systems (e.g., ArcB, PhoQ). The DNA-binding protein Dps that is generally regarded as cytoplasmic was significantly enriched in the outer membrane for all growth conditions examined, suggestive of moonlighting activities. These observations imply the existence of unknown transport mechanisms and novel functions for a subset of Salmonella proteins. Overall, this work provides a catalog of experimentally verified subcellular protein location for Salmonella and a framework for further investigations using computational modeling.

  9. Robust Road Condition Detection System Using In-Vehicle Standard Sensors

    Directory of Open Access Journals (Sweden)

    Juan Jesús Castillo Aguilar

    2015-12-01

    Full Text Available The appearance of active safety systems, such as Anti-lock Braking System, Traction Control System, Stability Control System, etc., represents a major evolution in road safety. In the automotive sector, the term vehicle active safety systems refers to those whose goal is to help avoid a crash or to reduce the risk of having an accident. These systems safeguard us, being in continuous evolution and incorporating new capabilities continuously. In order for these systems and vehicles to work adequately, they need to know some fundamental information: the road condition on which the vehicle is circulating. This early road detection is intended to allow vehicle control systems to act faster and more suitably, thus obtaining a substantial advantage. In this work, we try to detect the road condition the vehicle is being driven on, using the standard sensors installed in commercial vehicles. Vehicle models were programmed in on-board systems to perform real-time estimations of the forces of contact between the wheel and road and the speed of the vehicle. Subsequently, a fuzzy logic block is used to obtain an index representing the road condition. Finally, an artificial neural network was used to provide the optimal slip for each surface. Simulations and experiments verified the proposed method.

  10. Competitive association binding kinetic assays: a new tool to detect two different binding orientations of a ligand to its target protein under distinct conditions?

    Science.gov (United States)

    Wittmann, Hans-Joachim; Strasser, Andrea

    2017-06-01

    Within the last years, for several ligands, binding to G protein-coupled receptors or other target proteins, a binding of the ligand in two different orientations is described. One appropriate experimental technique to detect two different binding orientations is the crystallization of the ligand-protein-complex, but crystallization and subsequent X-ray analysis do not belong to the routine methods. By traditional competitive radioligand equilibrium binding assays, it is not possible to detect or to distinguish between two different binding orientations, but there is a possibility to identify two different binding orientations by performing kinetic competitive radioligand-binding assays. To study the limitations of this new technique, the related differential equations were defined and solved numerically for 8 different sets of rate constants, also considering an experimental error up to ~10%. In principal, the kinetic competitive radioligand binding assay is a suitable technique to detect two different ligand binding orientations. However, the present study shows that this is only possible under distinct conditions: (1) the rate constants of dissociation for both binding orientations of the cold ligand should at least be > 10-fold different to each other and (2) the experimental error should be as small as possible. Although there are some limitations for the experimental usability of this method, it is worthwhile to perform kinetic competitive binding assays, especially if there are hints for two binding orientations of a ligand, e.g. based on molecular modelling studies.

  11. Effect of measurement conditions on three-dimensional roughness values, and development of measurement standard

    International Nuclear Information System (INIS)

    Fabre, A; Brenier, B; Raynaud, S

    2011-01-01

    Friction or corrosion behaviour, fatigue lifetime for mechanical components are influenced by their boundary and subsurface properties. The surface integrity is studied on mechanical component in order to improve the service behaviour of them. Roughness is one of the main geometrical properties, which is to be qualified and quantified. Components can be obtained using a complex process: forming, machining and treatment can be combined to realize parts with complex shape. Then, three-dimensional roughness is needed to characterize these parts with complex shape and textured surface. With contact or non-contact measurements (contact stylus, confocal microprobe, interferometer), three-dimensional roughness is quantified using the calculation of pertinent parameters defined by the international standard PR EN ISO 25178-2:2008. An analysis will identify the influence of measurement conditions on three-dimensional parameters. The purpose of this study is to analyse the variation of roughness results using contact stylus or optical apparatus. The second aim of this work is to develop a measurement standard well adapted to qualify the contact and non-contact apparatus.

  12. Standard test method for damage to contacting solid surfaces under fretting conditions

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2010-01-01

    1.1 This test method covers the studying or ranking the susceptibility of candidate materials to fretting corrosion or fretting wear for the purposes of material selection for applications where fretting corrosion or fretting wear can limit serviceability. 1.2 This test method uses a tribological bench test apparatus with a mechanism or device that will produce the necessary relative motion between a contacting hemispherical rider and a flat counterface. The rider is pressed against the flat counterface with a loading mass. The test method is intended for use in room temperature air, but future editions could include fretting in the presence of lubricants or other environments. 1.3 The purpose of this test method is to rub two solid surfaces together under controlled fretting conditions and to quantify the damage to both surfaces in units of volume loss for the test method. 1.4 The values stated in SI units are to be regarded as standard. No other units of measurement are included in this standard. 1.5...

  13. Effect of measurement conditions on three-dimensional roughness values, and development of measurement standard

    Energy Technology Data Exchange (ETDEWEB)

    Fabre, A; Brenier, B [Arts et Metiers ParisTech, MecaSurf Laboratory, 2, Cours des Arts et Metiers, 13617 Aix-en-Provence (France); Raynaud, S, E-mail: agnes.fabre@ensam.eu [INSA Lyon, MIP2 Laboratory, 27 Avenue Jean Capelle, Bat Jacquard, 69100 Villeurbanne (France)

    2011-08-19

    Friction or corrosion behaviour, fatigue lifetime for mechanical components are influenced by their boundary and subsurface properties. The surface integrity is studied on mechanical component in order to improve the service behaviour of them. Roughness is one of the main geometrical properties, which is to be qualified and quantified. Components can be obtained using a complex process: forming, machining and treatment can be combined to realize parts with complex shape. Then, three-dimensional roughness is needed to characterize these parts with complex shape and textured surface. With contact or non-contact measurements (contact stylus, confocal microprobe, interferometer), three-dimensional roughness is quantified using the calculation of pertinent parameters defined by the international standard PR EN ISO 25178-2:2008. An analysis will identify the influence of measurement conditions on three-dimensional parameters. The purpose of this study is to analyse the variation of roughness results using contact stylus or optical apparatus. The second aim of this work is to develop a measurement standard well adapted to qualify the contact and non-contact apparatus.

  14. The development of standard samples with a defined number of antigen-specific T cells to harmonize T cell assays: a proof-of-principle study.

    Science.gov (United States)

    Singh, Satwinder Kaur; Tummers, Bart; Schumacher, Ton N; Gomez, Raquel; Franken, Kees L M C; Verdegaal, Els M; Laske, Karoline; Gouttefangeas, Cécile; Ottensmeier, Christian; Welters, Marij J P; Britten, Cedrik M; van der Burg, Sjoerd H

    2013-03-01

    The validation of assays that quantify antigen-specific T cell responses is critically dependent on cell samples that contain clearly defined measurable numbers of antigen-specific T cells. An important requirement is that such cell samples are handled and analyzed in a comparable fashion to peripheral blood mononuclear cells (PBMC). We performed a proof-of-principle study to show that retrovirally TCR-transduced T cells spiked at defined numbers in autologous PBMC can be used as standard samples for HLA/peptide multimer staining. NY-ESO-1157-165-specific, TCR-transduced CD8+ T cell batches were successfully generated from PBMC of several HLA-A*0201 healthy donors, purified by magnetic cell sorting on the basis of HLA tetramer (TM) staining and expanded with specific antigen in vitro. When subsequently spiked into autologous PBMC, the detection of these CD3+CD8+TM+ T cells was highly accurate with a mean accuracy of 91.6 %. The standard cells can be preserved for a substantial period of time in liquid nitrogen. Furthermore, TM staining of fresh and cryopreserved standard samples diluted at decreasing concentrations into autologous cryopreserved unspiked PBMC revealed that the spiked CD3+CD8+TM+ T cells could be accurately detected at all dilutions in a linear fashion with a goodness-of-fit of over 0.99 at a frequency of at least 0.02 % among the CD3+CD8+ T cell population. Notably, the CD3+CD8+TM+ cells of the standard samples were located exactly within the gates used to analyze patient samples and displayed a similar scatter pattern. The performance of the cryopreserved standard samples in the hands of 5 external investigators was good with an inter-laboratory variation of 32.9 % and the doubtless identification of one outlier.

  15. Mercury-Cadmium-Telluride Focal Plane Array Performance Under Non-Standard Operating Conditions

    Science.gov (United States)

    Richardson, Brandon S.; Eastwood, Michael L.; Bruce, Carl F.; Green, Robert O.; Coles, J. B.

    2011-01-01

    This paper highlights a new technique that allows the Teledyne Scientific & Imaging LLC TCM6604A Mercury-Cadmium-Telluride (MCT) Focal Plane Array (FPA) to operate at room temperature. The Teledyne MCT FPA has been a standard in Imaging Spectroscopy since its creation in the 1980's. This FPA has been used in applications ranging from space instruments such as CRISM, M3 and ARTEMIS to airborne instruments such as MaRS and the Next Generation AVIRIS Instruments1. Precise focal plane alignment is always a challenge for such instruments. The current FPA alignment process results in multiple cold cycles requiring week-long durations, thereby increasing the risk and cost of a project. These alignment cycles are necessary because optimal alignment is approached incrementally and can only be measured with the FPA and Optics at standard operating conditions, requiring a cold instrument. Instruments using this FPA are normally cooled to temperatures below 150K for the MCT FPA to properly function. When the FPA is run at higher temperatures the dark current increases saturating the output. This paper covers the prospect of warm MCT FPA operation from a theoretical and experimental perspective. We discuss the empirical models and physical laws that govern MCT material properties and predict the optimal settings that will result in the best MCT PA performance at 300K. Theoretical results are then calculated for the proposed settings. We finally present the images and data obtained using the actual system with the warm MCT FPA settings. The paper concludes by emphasizing the strong positive correlation between the measured values and the theoretical results.

  16. A seismic analysis of Korean standard PWR fuels under transition core conditions

    International Nuclear Information System (INIS)

    Kim, Hyeong Koo; Park, Nam Kyu; Jang, Young Ki; Kim, Jae Ik; Kim, Kyu Tae

    2005-01-01

    The PLUS7 fuel is developed to achieve higher thermal performance, burnup and more safety margin than the conventional fuel used in the Korean Standard Nuclear Plants (KSNPs) and to sustain structural integrity under increased seismic requirement in Korea. In this study, a series of seismic analysis have been performed in order to evaluate the structural integrity of fuel assemblies associated with seismic loads in the KSNPs under transition core conditions replacing the Guardian fuel, which is a resident fuel in the KSNP reactors, with the PLUS7 fuel. For the analysis, transition core seismic models have been developed, based on the possible fuel loading patterns. And the maximum impact forces on the spacer grid and various stresses acting on the fuel components have been evaluated and compared with the through-grid strength of spacer grids and the stress criteria specified in the ASME code for each fuel component, respectively. Then three noticeable parameters regarding as important parameters governing fuel assembly dynamic behavior are evaluated to clarify their effects on the fuel impact and stress response. As a result of the study, it has been confirmed that both the PLUS7 and the Guardian fuel sustain their structural integrity under the transition core condition. And when the damping ratio is constant, increasing the natural frequency of fuel assembly results in a decrease in impact force. The fuel assembly flexural stiffness has an effect increasing the stress of fuel assembly, but not the impact force. And the spacer grid stiffness is directly related with the impact force response. (author)

  17. Activity of Medicinal Plant Extracts on Multiplication of Mycobacterium tuberculosis under Reduced Oxygen Conditions Using Intracellular and Axenic Assays.

    Science.gov (United States)

    Bhatter, Purva D; Gupta, Pooja D; Birdi, Tannaz J

    2016-01-01

    Aim. Test the activity of selected medicinal plant extracts on multiplication of Mycobacterium tuberculosis under reduced oxygen concentration which represents nonreplicating conditions. Material and Methods. Acetone, ethanol and aqueous extracts of the plants Acorus calamus L. (rhizome), Ocimum sanctum L. (leaf), Piper nigrum L. (seed), and Pueraria tuberosa DC. (tuber) were tested on Mycobacterium tuberculosis H37Rv intracellularly using an epithelial cell (A549) infection model. The extracts found to be active intracellularly were further studied axenically under reducing oxygen concentrations. Results and Conclusions. Intracellular multiplication was inhibited ≥60% by five of the twelve extracts. Amongst these 5 extracts, in axenic culture, P. nigrum (acetone) was active under aerobic, microaerophilic, and anaerobic conditions indicating presence of multiple components acting at different levels and P. tuberosa (aqueous) showed bactericidal activity under microaerophilic and anaerobic conditions implying the influence of anaerobiosis on its efficacy. P. nigrum (aqueous) and A. calamus (aqueous and ethanol) extracts were not active under axenic conditions but only inhibited intracellular growth of Mycobacterium tuberculosis, suggesting activation of host defense mechanisms to mediate bacterial killing rather than direct bactericidal activity.

  18. Experimental study of the performance of intumescent coatings exposed to standard and non-standard fire conditions

    DEFF Research Database (Denmark)

    Lucherini, Andrea; Giuliani, Luisa; Jomaas, Grunde

    2018-01-01

    Three different experimental setups corresponding to three different fire scenarios were used to investigate how different heating conditions and heating rates affect the behaviour of two different thin intumescent coatings (a solvent-based and a water-based paint). Coated steel samples were...... to four critical points: activation, end of reaction, binder exhaustion and steel austenitization point. The results also showed that the water-based paint performed better at low heating rates, while the tested solvent-based paint performed better at high heating rates and did not activate or provide...

  19. Radioreceptor assays

    International Nuclear Information System (INIS)

    Lapka, R.

    1985-01-01

    Radioreceptor assay (RRA) is an analytical method using the specific interaction of some pharmaceuticals and endogenic substances (ligands) with specific receptors present in certin tissues of living organisms. RRA uses the principle of isotope dilution. The method is described in detail of the preparation of receptors, samples and radioligands, conditions of incubation, the separation of free and bound radioligand, and the mathematical evaluation of RRA. The sensitivity of RRA is measured in units to tens of pg. The specificity of RRA relates to a group of substances with similar pharmacological effect. RRA may be used for identifying neuroleptics, antidepressants, anxiolytics, ergot alkaloids, beta blockers, anticholinergic drugs, certain hormones and neuropeptides. (M.D.)

  20. Carbon dioxide and ethanol release from champagne glasses, under standard tasting conditions.

    Science.gov (United States)

    Liger-Belair, Gérard; Beaumont, Fabien; Bourget, Marielle; Pron, Hervé; Parvitte, Bertrand; Zéninari, Virginie; Polidori, Guillaume; Cilindre, Clara

    2012-01-01

    A simple glass of champagne or sparkling wine may seem like the acme of frivolity to most people, but in fact, it may rather be considered as a fantastic playground for any fluid physicist or physicochemist. In this chapter, results obtained concerning various steps where the CO₂ molecule plays a role (from its ingestion in the liquid phase during the fermentation process to its progressive release in the headspace above the tasting glass) are gathered and synthesized to propose a self-consistent and global overview of how gaseous and dissolved CO₂ impact champagne and sparkling wine science. Some recent investigations, conducted through laser tomography techniques, on ascending bubbles and ascending-bubble-driven flow patterns found in champagne glasses are reported, which illustrate the fine interplay between ascending bubbles and the fluid around under standard tasting conditions. The simultaneous monitoring of gaseous CO₂ and ethanol in the headspace of both a flute and a coupe filled with champagne was reported, depending on whether or not the glass shows effervescence. Both gaseous CO₂ and ethanol were found to be enhanced by the presence of ascending bubbles, thus confirming the close link between ascending bubbles, ascending-bubble-driven flow patterns, and the release of gaseous CO₂ and volatile organic compounds. Copyright © 2012 Elsevier Inc. All rights reserved.

  1. A new reliable, transposable and cost-effective assay for absolute quantification of total plasmatic bevacizumab by LC-MS/MS in human plasma comparing two internal standard calibration approaches.

    Science.gov (United States)

    Legeron, Rachel; Xuereb, Fabien; Chaignepain, Stephane; Gadeau, Alain-Pierre; Claverol, Stephane; Dupuy, Jean-William; Djabarouti, Sarah; Couffinhal, Thierry; Schmitter, Jean-Marie; Breilh, Dominique

    2017-12-01

    The quantification of monoclonal antibodies (mAbs) such as bevacizumab, a recombinant humanized immunoglobulin G1 (hIgG1), in biological fluids, is an essential prerequisite to any pharmacokinetic preclinical and clinical study. To date, reference techniques used to quantify mAbs rely on enzyme-linked immunosorbent assay (ELISA) lacking specificity. Furthermore, the commercially available ELISA kit to quantify bevacizumab in human plasma only assesses the free fraction of the drug. However, the conditions of storage and analysis of plasma samples could alter the physiological equilibrium between the free, bound and partially bound forms of bevacizumab and this could result in over- or underestimation of drug concentration. We developed a new assay for absolute quantification of total fraction of bevacizumab by liquid chromatography tandem mass spectrometry (LC-MS/MS) basing identification and quantification of bevacizumab on two specific peptides. In this report we compare our assay with two internal standard (IS) calibration approaches: one using a different human mAb (Trastuzumab) and the other using a stable isotope labeled specific peptide. After enrichment by affinity chromatography on protein A and concentration by ultrafiltration, human plasma samples were proteolyzed by trypsin. Linearity was established from 12.5 to 500μg/mL with an interday accuracy ranging from 101.7 to 110.6% and precision from 7.0% to 9.9%. This study demonstrates the importance of the choice of the IS in quantifying bevacizumab in human plasma and highlights the difficulty of reaching a reliable proteolysis with a sufficient recovery. We developed a reliable and cost-effective LC-MS/MS method to quantify total plasmatic fraction of bevacizumab in human plasma. Through our development we proposed a generic methodology easily transposable to quantify all IgG1 subclass very useful for clinical pharmacokinetics studies. Copyright © 2017 Elsevier B.V. All rights reserved.

  2. Compensation for the Effects of Ambient Conditions on the Calibration of Multi-Capillary Pressure Drop Standards

    Directory of Open Access Journals (Sweden)

    Colard S

    2014-12-01

    Full Text Available Cigarette draw resistance and filter pressure drop (PD are both major physical parameters for the tobacco industry. Therefore these parameters must be measured reliably. For these measurements, specific equipment calibrated with PD transfer standards is used. Each transfer standard must have a known and stable PD value, such standards usually being composed of several capillary tubes associated in parallel. However, PD values are modified by ambient conditions during calibration of such standards, i.e. by temperature and relative humidity (RH of air, and atmospheric pressure. In order to reduce the influence of these ambient factors, a simplified model was developed for compensating the effects of ambient conditions on the calibration of multi-capillary PD standards.

  3. A Biosensor-Based Leaf Punch Assay for Glutamine Correlates to Symbiotic Nitrogen Fixation Measurements in Legumes to Permit Rapid Screening of Rhizobia Inoculants under Controlled Conditions.

    Science.gov (United States)

    Thilakarathna, Malinda S; Moroz, Nicholas; Raizada, Manish N

    2017-01-01

    Legumes are protein sources for billions of humans and livestock. These traits are enabled by symbiotic nitrogen fixation (SNF), whereby root nodule-inhabiting rhizobia bacteria convert atmospheric nitrogen (N) into usable N. Unfortunately, SNF rates in legume crops suffer from undiagnosed incompatible/suboptimal interactions between crop varieties and rhizobia strains. There are opportunities to test much large numbers of rhizobia strains if cost/labor-effective diagnostic tests become available which may especially benefit researchers in developing countries. Inside root nodules, fixed N from rhizobia is assimilated into amino acids including glutamine (Gln) for export to shoots as the major fraction (amide-exporting legumes) or as the minor fraction (ureide-exporting legumes). Here, we have developed a new leaf punch based technique to screen rhizobia inoculants for SNF activity following inoculation of both amide exporting and ureide exporting legumes. The assay is based on measuring Gln output using the GlnLux biosensor, which consists of Escherichia coli cells auxotrophic for Gln and expressing a constitutive lux operon. Subsistence farmer varieties of an amide exporter (lentil) and two ureide exporters (cowpea and soybean) were inoculated with different strains of rhizobia under controlled conditions, then extracts of single leaf punches were incubated with GlnLux cells, and light-output was measured using a 96-well luminometer. In the absence of external N and under controlled conditions, the results from the leaf punch assay correlated with 15 N-based measurements, shoot N percentage, and shoot total fixed N in all three crops. The technology is rapid, inexpensive, high-throughput, requires minimum technical expertise and very little tissue, and hence is relatively non-destructive. We compared and contrasted the benefits and limitations of this novel diagnostic assay to methods.

  4. A Biosensor-Based Leaf Punch Assay for Glutamine Correlates to Symbiotic Nitrogen Fixation Measurements in Legumes to Permit Rapid Screening of Rhizobia Inoculants under Controlled Conditions

    Directory of Open Access Journals (Sweden)

    Malinda S. Thilakarathna

    2017-10-01

    Full Text Available Legumes are protein sources for billions of humans and livestock. These traits are enabled by symbiotic nitrogen fixation (SNF, whereby root nodule-inhabiting rhizobia bacteria convert atmospheric nitrogen (N into usable N. Unfortunately, SNF rates in legume crops suffer from undiagnosed incompatible/suboptimal interactions between crop varieties and rhizobia strains. There are opportunities to test much large numbers of rhizobia strains if cost/labor-effective diagnostic tests become available which may especially benefit researchers in developing countries. Inside root nodules, fixed N from rhizobia is assimilated into amino acids including glutamine (Gln for export to shoots as the major fraction (amide-exporting legumes or as the minor fraction (ureide-exporting legumes. Here, we have developed a new leaf punch based technique to screen rhizobia inoculants for SNF activity following inoculation of both amide exporting and ureide exporting legumes. The assay is based on measuring Gln output using the GlnLux biosensor, which consists of Escherichia coli cells auxotrophic for Gln and expressing a constitutive lux operon. Subsistence farmer varieties of an amide exporter (lentil and two ureide exporters (cowpea and soybean were inoculated with different strains of rhizobia under controlled conditions, then extracts of single leaf punches were incubated with GlnLux cells, and light-output was measured using a 96-well luminometer. In the absence of external N and under controlled conditions, the results from the leaf punch assay correlated with 15N-based measurements, shoot N percentage, and shoot total fixed N in all three crops. The technology is rapid, inexpensive, high-throughput, requires minimum technical expertise and very little tissue, and hence is relatively non-destructive. We compared and contrasted the benefits and limitations of this novel diagnostic assay to methods.

  5. Injury Rates in Age-Only Versus Age-and-Weight Playing Standard Conditions in American Youth Football

    Science.gov (United States)

    Kerr, Zachary Y.; Marshall, Stephen W.; Simon, Janet E.; Hayden, Ross; Snook, Erin M.; Dodge, Thomas; Gallo, Joseph A.; Valovich McLeod, Tamara C.; Mensch, James; Murphy, Joseph M.; Nittoli, Vincent C.; Dompier, Thomas P.; Ragan, Brian; Yeargin, Susan W.; Parsons, John T.

    2015-01-01

    Background: American youth football leagues are typically structured using either age-only (AO) or age-and-weight (AW) playing standard conditions. These playing standard conditions group players by age in the former condition and by a combination of age and weight in the latter condition. However, no study has systematically compared injury risk between these 2 playing standards. Purpose: To compare injury rates between youth tackle football players in the AO and AW playing standard conditions. Study Design: Cohort study; Level of evidence, 2. Methods: Athletic trainers evaluated and recorded injuries at each practice and game during the 2012 and 2013 football seasons. Players (age, 5-14 years) were drawn from 13 recreational leagues across 6 states. The sample included 4092 athlete-seasons (AW, 2065; AO, 2027) from 210 teams (AW, 106; O, 104). Injury rate ratios (RRs) with 95% CIs were used to compare the playing standard conditions. Multivariate Poisson regression was used to estimate RRs adjusted for residual effects of age and clustering by team and league. There were 4 endpoints of interest: (1) any injury, (2) non–time loss (NTL) injuries only, (3) time loss (TL) injuries only, and (4) concussions only. Results: Over 2 seasons, the cohort accumulated 1475 injuries and 142,536 athlete-exposures (AEs). The most common injuries were contusions (34.4%), ligament sprains (16.3%), concussions (9.6%), and muscle strains (7.8%). The overall injury rate for both playing standard conditions combined was 10.3 per 1000 AEs (95% CI, 9.8-10.9). The TL injury, NTL injury, and concussion rates in both playing standard conditions combined were 3.1, 7.2, and 1.0 per 1000 AEs, respectively. In multivariate Poisson regression models controlling for age, team, and league, no differences were found between playing standard conditions in the overall injury rate (RRoverall, 1.1; 95% CI, 0.4-2.6). Rates for the other 3 endpoints were also similar (RRNTL, 1.1 [95% CI, 0

  6. 78 FR 73112 - Monitoring System Conditions-Transmission Operations Reliability Standards; Interconnection...

    Science.gov (United States)

    2013-12-05

    ... complement the TOP Standards, have the goal of ensuring that the bulk electric system is planned and operated... pertain to the coordinated efforts to operate the bulk electric system in a reliable manner during real... System Operating Limits (SOLs).\\5\\ The provisions in the proposed TOP Reliability Standards that require...

  7. Autofluorescence of atmospheric bioaerosols - Biological standard particles and the influence of environmental conditions

    Science.gov (United States)

    Pöhlker, Christopher; Huffman, J. Alex; Förster, Jan-David; Pöschl, Ulrich

    2013-04-01

    standard bioparticles (pollen, fungal spores, and bacteria) as well as atmospherically relevant chemical substances. We addressed the sensitivity and selectivity of autofluorescence based online techniques. Moreover, we investigated the influence of environmental conditions, such as relative humidity and oxidizing agents in the atmosphere, on the autofluorescence signature of standard bioparticles. Our results will support the molecular understanding and quantitative interpretation of data obtained by real-time FBAP instrumentation [5,6]. [1] Elbert, W., Taylor, P. E., Andreae, M. O., & Pöschl, U. (2007). Atmos. Chem. Phys., 7, 4569-4588. [2] Huffman, J. A., Treutlein, B., & Pöschl, U. (2010). Atmos. Chem. Phys., 10, 3215-3233. [3] Pöschl, U., et al. (2010). Science, 329, 1513-1516. [4] Lakowicz, J., Principles of fluorescence spectroscopy, Plenum publishers, New York, 1999. [5] Pöhlker, C., Huffman, J. A., & Pöschl, U., (2012). Atmos. Meas. Tech., 5, 37-71. [6] Pöhlker, C., Huffman, J. A., Förster J.-D., & Pöschl, U., (2012) in preparation.

  8. Standard nomenclature and methods for describing the condition of pavements draft TRH 6

    CSIR Research Space (South Africa)

    Curtayne, PC

    2009-01-26

    Full Text Available The need for describing the condition of pavements occurs frequently in highway engineering. Accurate descriptions are a prerequisite for establishing procedures with which to evaluate the various aspects of the pavement condition. A variety...

  9. Quantitative monitoring of HCMV DNAlactia in human milk by real time PCR assay: Implementation of internal control contributes to standardization and quality control.

    Science.gov (United States)

    Hartleif, Steffen; Göhring, Katharina; Goelz, Rangmar; Jahn, Gerhard; Hamprecht, Klaus

    2016-11-01

    For cytomegalovirus screening of breastfeeding mothers of preterm infants under risk, we present a rapid, quantitative real-time PCR protocol using the hybridization format of the viral gB target region. For quantification, we used an external gB fragment cloned into a vector system. For standardization, we created an internal control-plasmid by site-directed mutagenesis with an exchange of 9 nucleotides. Spiked with internal control, patient wildtype amplicons could be discriminated from internal controls by hybridization probes using two-channel fluorescence detection. Potential bias of formerly reported false nucleotide sequence data of gB-hybridization probes was excluded. Using this approach, we could demonstrate excellent analytical performance and high reproducibility of HCMV detection during lactation. This assay shows very good correlation with a commercial quantitative HCMV DNA PCR and may help to identify rapidly HCMV shedding mothers of very low birth weight preterm infants to prevent HCMV transmission. On the other hand, negative DNA amplification results allow feeding of milk samples of seropositive mothers to their preterm infants under risk (<30 weeks of gestational age, <1000g birth weight) during the onset and late stage of HCMV shedding during lactation. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Different Roles for Honey Bee Mushroom Bodies and Central Complex in Visual Learning of Colored Lights in an Aversive Conditioning Assay.

    Science.gov (United States)

    Plath, Jenny A; Entler, Brian V; Kirkerud, Nicholas H; Schlegel, Ulrike; Galizia, C Giovanni; Barron, Andrew B

    2017-01-01

    The honey bee is an excellent visual learner, but we know little about how and why it performs so well, or how visual information is learned by the bee brain. Here we examined the different roles of two key integrative regions of the brain in visual learning: the mushroom bodies and the central complex. We tested bees' learning performance in a new assay of color learning that used electric shock as punishment. In this assay a light field was paired with electric shock. The other half of the conditioning chamber was illuminated with light of a different wavelength and not paired with shocks. The unrestrained bee could run away from the light stimulus and thereby associate one wavelength with punishment, and the other with safety. We compared learning performance of bees in which either the central complex or mushroom bodies had been transiently inactivated by microinjection of the reversible anesthetic procaine. Control bees learned to escape the shock-paired light field and to spend more time in the safe light field after a few trials. When ventral lobe neurons of the mushroom bodies were silenced, bees were no longer able to associate one light field with shock. By contrast, silencing of one collar region of the mushroom body calyx did not alter behavior in the learning assay in comparison to control treatment. Bees with silenced central complex neurons did not leave the shock-paired light field in the middle trials of training, even after a few seconds of being shocked. We discussed how mushroom bodies and the central complex both contribute to aversive visual learning with an operant component.

  11. Different Roles for Honey Bee Mushroom Bodies and Central Complex in Visual Learning of Colored Lights in an Aversive Conditioning Assay

    Directory of Open Access Journals (Sweden)

    Jenny A. Plath

    2017-05-01

    Full Text Available The honey bee is an excellent visual learner, but we know little about how and why it performs so well, or how visual information is learned by the bee brain. Here we examined the different roles of two key integrative regions of the brain in visual learning: the mushroom bodies and the central complex. We tested bees' learning performance in a new assay of color learning that used electric shock as punishment. In this assay a light field was paired with electric shock. The other half of the conditioning chamber was illuminated with light of a different wavelength and not paired with shocks. The unrestrained bee could run away from the light stimulus and thereby associate one wavelength with punishment, and the other with safety. We compared learning performance of bees in which either the central complex or mushroom bodies had been transiently inactivated by microinjection of the reversible anesthetic procaine. Control bees learned to escape the shock-paired light field and to spend more time in the safe light field after a few trials. When ventral lobe neurons of the mushroom bodies were silenced, bees were no longer able to associate one light field with shock. By contrast, silencing of one collar region of the mushroom body calyx did not alter behavior in the learning assay in comparison to control treatment. Bees with silenced central complex neurons did not leave the shock-paired light field in the middle trials of training, even after a few seconds of being shocked. We discussed how mushroom bodies and the central complex both contribute to aversive visual learning with an operant component.

  12. Assay of anti-cancer drugs in tissue culture: conditions affecting their ability to incorporate 3H-leucine after drug treatment.

    Science.gov (United States)

    Freshney, R I; Paul, J; Kane, I M

    1975-01-01

    An attempt has been made to construct an assay potentially suitable for use with primary cultures of human tumours to measure the survival of exponentially growing monolayer cultures after exposure to anti-neoplastic drugs. Cell survival was assessed using their protein synthetic capacity after removal of drugs. HeLa cells were employed to avoid the ingerent variability and heterogeneity of primary cultures from human tumours, and an assay has been constructed using microtitration trays to provide large numbers of replicate cultures without the requirement of a large number cells. An increase in the duration of the exposure to drug increased sensitivity in nearly all cases examined. Similarly, an increase in the period of culture following drug removal produced increased sensitivity to alkylating agents but allowed recovery from exposure to certain cycle-dependent drugs. Some of the drugs used were shown to be unstable under culture conditions and vinblastine was actively metabolized, although this instability was not necessarily reflected in the time course of the drug's effect. Mustine sensitivity was shown to be reduced by an increase in cell density at a level where density limitation of 3H-thymidine incorporation becomes apparent. These variations and possible methods of minimizing their effects are discussed.

  13. Standardization and validation of real time PCR assays for the diagnosis of histoplasmosis using three molecular targets in an animal model.

    Directory of Open Access Journals (Sweden)

    Luisa F López

    Full Text Available Histoplasmosis is considered one of the most important endemic and systemic mycoses worldwide. Until now few molecular techniques have been developed for its diagnosis. The aim of this study was to develop and evaluate three real time PCR (qPCR protocols for different protein-coding genes (100-kDa, H and M antigens using an animal model. Fresh and formalin-fixed and paraffin-embedded (FFPE lung tissues from BALB/c mice inoculated i.n. with 2.5x106 Histoplasma capsulatum yeast or PBS were obtained at 1, 2, 3, 4, 8, 12 and 16 weeks post-infection. A collection of DNA from cultures representing different clades of H. capsulatum (30 strains and other medically relevant pathogens (36 strains of related fungi and Mycobacterium tuberculosis were used to analyze sensitivity and specificity. Analytical sensitivity and specificity were 100% when DNAs from the different strains were tested. The highest fungal burden occurred at first week post-infection and complete fungal clearance was observed after the third week; similar results were obtained when the presence of H. capsulatum yeast cells was demonstrated in histopathological analysis. In the first week post-infection, all fresh and FFPE lung tissues from H. capsulatum-infected animals were positive for the qPCR protocols tested except for the M antigen protocol, which gave variable results when fresh lung tissue samples were analyzed. In the second week, all qPCR protocols showed variable results for both fresh and FFPE tissues. Samples from the infected mice at the remaining times post-infection and uninfected mice (controls were negative for all protocols. Good agreement was observed between CFUs, histopathological analysis and qPCR results for the 100-kDa and H antigen protocols. We successfully standardized and validated three qPCR assays for detecting H. capsulatum DNA in fresh and FFPE tissues, and conclude that the 100-kDa and H antigen molecular assays are promising tests for diagnosing this

  14. Standardization and validation of real time PCR assays for the diagnosis of histoplasmosis using three molecular targets in an animal model.

    Science.gov (United States)

    López, Luisa F; Muñoz, César O; Cáceres, Diego H; Tobón, Ángela M; Loparev, Vladimir; Clay, Oliver; Chiller, Tom; Litvintseva, Anastasia; Gade, Lalitha; González, Ángel; Gómez, Beatriz L

    2017-01-01

    Histoplasmosis is considered one of the most important endemic and systemic mycoses worldwide. Until now few molecular techniques have been developed for its diagnosis. The aim of this study was to develop and evaluate three real time PCR (qPCR) protocols for different protein-coding genes (100-kDa, H and M antigens) using an animal model. Fresh and formalin-fixed and paraffin-embedded (FFPE) lung tissues from BALB/c mice inoculated i.n. with 2.5x106 Histoplasma capsulatum yeast or PBS were obtained at 1, 2, 3, 4, 8, 12 and 16 weeks post-infection. A collection of DNA from cultures representing different clades of H. capsulatum (30 strains) and other medically relevant pathogens (36 strains of related fungi and Mycobacterium tuberculosis) were used to analyze sensitivity and specificity. Analytical sensitivity and specificity were 100% when DNAs from the different strains were tested. The highest fungal burden occurred at first week post-infection and complete fungal clearance was observed after the third week; similar results were obtained when the presence of H. capsulatum yeast cells was demonstrated in histopathological analysis. In the first week post-infection, all fresh and FFPE lung tissues from H. capsulatum-infected animals were positive for the qPCR protocols tested except for the M antigen protocol, which gave variable results when fresh lung tissue samples were analyzed. In the second week, all qPCR protocols showed variable results for both fresh and FFPE tissues. Samples from the infected mice at the remaining times post-infection and uninfected mice (controls) were negative for all protocols. Good agreement was observed between CFUs, histopathological analysis and qPCR results for the 100-kDa and H antigen protocols. We successfully standardized and validated three qPCR assays for detecting H. capsulatum DNA in fresh and FFPE tissues, and conclude that the 100-kDa and H antigen molecular assays are promising tests for diagnosing this mycosis.

  15. Conditional conservatism and value relevance of financial reporting: A study in view of converging accounting standards

    NARCIS (Netherlands)

    Thijssen, Maximiliaan Willem Pierre; Iatridis, George Emmanuel

    2016-01-01

    This study examines the relationship between conditional conservatism and value relevance in the EU and US. Specifically, it investigates whether this relationship differs under US GAAP and IFRS compliance. In addition, this study examines the trend in value relevance, conditional conservatism and

  16. [Prediction of the total Japanese cedar pollen counts based on male flower-setting conditions of standard trees].

    Science.gov (United States)

    Yuta, Atsushi; Ukai, Kotaro; Sakakura, Yasuo; Tani, Hideshi; Matsuda, Fukiko; Yang, Tian-qun; Majima, Yuichi

    2002-07-01

    We made a prediction of the Japanese cedar (Cryptomeria japonica) pollen counts at Tsu city based on male flower-setting conditions of standard trees. The 69 standard trees from 23 kinds of clones, planted at Mie Prefecture Science and Technology Promotion Center (Hakusan, Mie) in 1964, were selected. Male flower-setting conditions for 276 faces (69 trees x 4 points of the compass) were scored from 0 to 3. The average of scores and total pollen counts from 1988 to 2000 was analyzed. As the results, the average scores from standard trees and total pollen counts except two mass pollen-scattered years in 1995 and 2000 had a positive correlation (r = 0.914) by linear function. On the mass pollen-scattered years, pollen counts were influenced from the previous year. Therefore, the score of the present year minus that of the previous year were used for analysis. The average scores from male flower-setting conditions and pollen counts had a strong positive correlation (r = 0.994) when positive scores by taking account of the previous year were analyzed. We conclude that prediction of pollen counts are possible based on the male flower-setting conditions of standard trees.

  17. 42 CFR 433.123 - Notification of changes in system requirements, performance standards or other conditions for...

    Science.gov (United States)

    2010-10-01

    ... 42 Public Health 4 2010-10-01 2010-10-01 false Notification of changes in system requirements, performance standards or other conditions for approval or reapproval. 433.123 Section 433.123 Public Health... ASSISTANCE PROGRAMS STATE FISCAL ADMINISTRATION Mechanized Claims Processing and Information Retrieval...

  18. A Guide for Developing Standard Operating Job Procedures for the Sludge Conditioning & Dewatering Process Wastewater Treatment Facility. SOJP No. 11.

    Science.gov (United States)

    Schwing, Carl M.

    This guide describes standard operating job procedures for the sludge conditioning and dewatering process of wastewater treatment facilities. In this process, sludge is treated with chemicals to make the sludge coagulate and give up its water more easily. The treated sludge is then dewatered using a vacuum filter. The guide gives step-by-step…

  19. Conditions for vacuum stability in an S{sub 3} extension of the Standard Model

    Energy Technology Data Exchange (ETDEWEB)

    Beltran, O Felix [Fac. de Cs. de la Electronica, BUAP, Apdo. Postal 542, Puebla, Pue. 72570 (Mexico); Mondragon, M [Instituto de Fisica, Universidad Nacional Autonoma de Mexico, Apdo. Postal 20-364, Mexico, D.F. 01000 (Mexico); RodrIguez-Jauregui, E, E-mail: ezequiel.rodriguez@correo.fisica.uson.m [Departamento de Fisica, UNISON, Apdo. Postal 1626, Hermosillo, Son. 83000 (Mexico)

    2009-06-01

    In this work we study the Higgs sector in the minimal S{sub 3} extension of the Standard Model. The S{sub 3} extended Standard Model, which has three Higgs doublets fields that belong to the three-dimensional reducible representation of the permutation group S{sub 3}, has naturally new phenomena: there are several Higgs bosons, charged, neutral and pseuodscalar ones, and more than one potential minimum. We analyzed the stability of the minimal S3 invariant extension of the Higgs potential and show that at tree-level, the potential minimum preserving electric charge and CP symmetries, when it exists, is the global one.

  20. Enzyme assays.

    Science.gov (United States)

    Brodelius, P E

    1991-02-01

    The past year or so has seen the development of new enzyme assays, as well as the improvement of existing ones. Assays are becoming more rapid and sensitive as a result of modifications such as amplification of the enzyme product(s). Recombinant DNA technology is now being recognized as a particularly useful tool in the search for improved assay systems.

  1. 24 CFR 886.307 - Physical condition standards; physical inspection requirements.

    Science.gov (United States)

    2010-04-01

    ..., subpart G. (b) Space and security. In addition to the standards in 24 CFR part 5, subpart G, the dwelling unit must have a living room, a kitchen area, and a bathroom. The dwelling unit must have at least one...) The unit shall contain suitable space to store, prepare and serve foods in a sanitary manner. A...

  2. Radioreceptor opioid assay

    International Nuclear Information System (INIS)

    Miller, R.J.; Chang, K.-J.

    1981-01-01

    A radioreceptor assay is described for assaying opioid drugs in biological fluids. The method enables the assay of total opioid activity, being specific for opioids as a class but lacking specificity within the class. A radio-iodinated opioid and the liquid test sample are incubated with an opiate receptor material. The percentage inhibition of the binding of the radio-iodinated compound to the opiate receptor is calculated and the opioid activity of the test liquid determined from a standard curve. Examples of preparing radio-iodinated opioids and assaying opioid activity are given. A test kit for the assay is described. Compared to other methods, this assay is cheap, easy and rapid. (U.K.)

  3. Wine yeast phenomics: A standardized fermentation method for assessing quantitative traits of Saccharomyces cerevisiae strains in enological conditions

    Science.gov (United States)

    Bernard, Margaux; Trujillo, Marine; Prodhomme, Duyên; Barbe, Jean-Christophe; Gibon, Yves; Marullo, Philippe

    2018-01-01

    This work describes the set up of a small scale fermentation methodology for measuring quantitative traits of hundreds of samples in an enological context. By using standardized screw cap vessels, the alcoholic fermentation kinetics of Saccharomyces cerevisiae strains were measured by following their weight loss over the time. This dispositive was coupled with robotized enzymatic assays for measuring metabolites of enological interest in natural grape juices. Despite the small volume used, kinetic parameters and fermentation end products measured are similar with those observed in larger scale vats. The vessel used also offers the possibility to assay 32 volatiles compounds using a headspace solid-phase micro-extraction coupled to gas chromatography and mass spectrometry. The vessel shaking applied strongly impacted most of the phenotypes investigated due to oxygen transfer occuring in the first hours of the alcoholic fermentation. The impact of grape must and micro-oxygenation was investigated illustrating some relevant genetic x environmental interactions. By phenotyping a wide panel of commercial wine starters in five grape juices, broad phenotypic correlations between kinetics and metabolic end products were evidentiated. Moreover, a multivariate analysis illustrates that some grape musts are more able than others to discriminate commercial strains since some are less robust to environmental changes. PMID:29351285

  4. Wine yeast phenomics: A standardized fermentation method for assessing quantitative traits of Saccharomyces cerevisiae strains in enological conditions.

    Science.gov (United States)

    Peltier, Emilien; Bernard, Margaux; Trujillo, Marine; Prodhomme, Duyên; Barbe, Jean-Christophe; Gibon, Yves; Marullo, Philippe

    2018-01-01

    This work describes the set up of a small scale fermentation methodology for measuring quantitative traits of hundreds of samples in an enological context. By using standardized screw cap vessels, the alcoholic fermentation kinetics of Saccharomyces cerevisiae strains were measured by following their weight loss over the time. This dispositive was coupled with robotized enzymatic assays for measuring metabolites of enological interest in natural grape juices. Despite the small volume used, kinetic parameters and fermentation end products measured are similar with those observed in larger scale vats. The vessel used also offers the possibility to assay 32 volatiles compounds using a headspace solid-phase micro-extraction coupled to gas chromatography and mass spectrometry. The vessel shaking applied strongly impacted most of the phenotypes investigated due to oxygen transfer occuring in the first hours of the alcoholic fermentation. The impact of grape must and micro-oxygenation was investigated illustrating some relevant genetic x environmental interactions. By phenotyping a wide panel of commercial wine starters in five grape juices, broad phenotypic correlations between kinetics and metabolic end products were evidentiated. Moreover, a multivariate analysis illustrates that some grape musts are more able than others to discriminate commercial strains since some are less robust to environmental changes.

  5. Research on a Valuation Standard and the Actual Condition About Security Management in PACS

    International Nuclear Information System (INIS)

    Jeong, Jae Ho; Son, Gi Gyeong; Kang, Hee Doo; Dong, Kyung Rae; Kweon, Dae Cheol; Kim, Hyun Soo

    2008-01-01

    This study is to prepare an evaluation standard about personal information protection and security management of a medical institution and to build up a grade standard of evaluation in PACS environment. We built up evaluation index based on 10 detailed items in four big categories (political security, technical security, data management security and physical security) by referring to ISO17799 (BS 7799), HIPPA (Health Insurance and Portability and Accountability Act of 1996) and domestic medical law. We have investigated at the thirty places where medical facility with the extracted security criteria and security evaluation index. Average score of physical security list, one of the big categories, was 18.5/20 (93%) at all medical institutions. Political security score was 18.5/30 (62%), data management security score was 12/20 (60%) and technical security score was 17.5/30 (58%). Therefore, security evaluation score was average 67 in 30 general hospitals, which was 4th level. The results showed that it is necessary to establish evaluation and management standard about personal information protection and security consciousness which are weak in PACS environment.

  6. Comparison of ESD and major organ absorbed doses of 5 year old standard guidekines and clinical exposure conditions

    Energy Technology Data Exchange (ETDEWEB)

    Kang, A Ram; Ahn, Sung Min [Dept. of Radiological Science, The Graduate School, Gachon University, Incheon (Korea, Republic of); Lee, In Ja [Dept. of Radiologic technology, Dongnam health University, Suwon (Korea, Republic of)

    2017-09-15

    Pediatrics are more sensibility to radiation than adults and because they are organs that are not completely grown, they have a life expectancy that can be adversely affected by exposure. Therefore, the management of exposure dose is more important than the case of adult. The purpose of this study was to determine the suitability of the 10 year old phantom for the 5 year old pediatric's recommendation and the incident surface dose, and to measure the organ absorbed dose. This study is compared the organ absorbed dose and the entrance surface dose in the clinical conditions at 5 and 10 years old pediatric. Clinical 5 year old condition was slightly higher than recommendation condition and 10 year old condition was very high. In addition, recommendation condition ESD was found to be 43% higher than the ESD of the 5 year old group and the ESD of the 10 year old group was 126% higher than that of the 5 year old group. The recommended ESD at 5 years old and the ESD according to clinical imaging conditions were 31.6%. There was no significant difference between the 5 year old recommended exposure conditions and the organ absorbed dose due to clinical exposure conditions, but there was a large difference between the Chest and Pelvic. However, it was found that there was a remarkable difference when comparing the organ absorbed dose by 10 year clinical exposure conditions. Therefore, more detailed standard exposure dose for the recommended dose of pediatric should be studied.

  7. Mathematical analysis of the Navier-Stokes equations with non standard boundary conditions

    Science.gov (United States)

    Tidriri, M. D.

    1995-01-01

    One of the major applications of the domain decomposition time marching algorithm is the coupling of the Navier-Stokes systems with Boltzmann equations in order to compute transitional flows. Another important application is the coupling of a global Navier-Stokes problem with a local one in order to use different modelizations and/or discretizations. Both of these applications involve a global Navier-Stokes system with nonstandard boundary conditions. The purpose of this work is to prove, using the classical Leray-Schauder theory, that these boundary conditions are admissible and lead to a well posed problem.

  8. Standard methods for maintaining adult Apis mellifera in cages under in vitro laboratory conditions

    NARCIS (Netherlands)

    Williams, G.R.; Alaux, C.; Costa, C.; Csaki, C.; Steen, van der J.J.M.

    2013-01-01

    Adult honey bees are maintained in vitro in laboratory cages for a variety of purposes. For example, researchers may wish to perform experiments on honey bees caged individually or in groups to study aspects of parasitology, toxicology, or physiology under highly controlled conditions, or they may

  9. 18 CFR 4.94 - Standard terms and conditions of exemption.

    Science.gov (United States)

    2010-04-01

    ... conditions of exemption. 4.94 Section 4.94 Conservation of Power and Water Resources FEDERAL ENERGY REGULATORY COMMISSION, DEPARTMENT OF ENERGY REGULATIONS UNDER THE FEDERAL POWER ACT LICENSES, PERMITS, EXEMPTIONS, AND DETERMINATION OF PROJECT COSTS Exemption of Small Conduit Hydroelectric Facilities § 4.94...

  10. Comparison study of judged clinical skills competence from standard setting ratings generated under different administration conditions.

    Science.gov (United States)

    Roberts, William L; Boulet, John; Sandella, Jeanne

    2017-12-01

    When the safety of the public is at stake, it is particularly relevant for licensing and credentialing exam agencies to use defensible standard setting methods to categorize candidates into competence categories (e.g., pass/fail). The aim of this study was to gather evidence to support change to the Comprehensive Osteopathic Medical Licensing-USA Level 2-Performance Evaluation standard setting design and administrative process. Twenty-two video recordings of candidates assessed for clinical competence were randomly selected from the 2014-2015 Humanistic domain test score distribution ranging from the highest to lowest quintile of performance. Nineteen panelists convened at the same site to receive training and practice prior to generating judgments of qualified or not qualified performance to each of the twenty videos. At the end of training, one panel remained onsite to complete their judgments and the second panel was released and given 1 week to observe the same twenty videos and complete their judgments offsite. The two one-sided test procedure established equivalence between panel group means at the 0.05 confidence level, controlling for rater errors within each panel group. From a practical cost-effective and administrative resource perspective, results from this study suggest it is possible to diverge from typical panel groups, who are sequestered the entire time onsite, to larger numbers of panelists who can make their judgments offsite with little impact on judged samples of qualified performance. Standard setting designs having panelists train together and then allowing those to provide judgments yields equivalent ratings and, ultimately, similar cut scores.

  11. Standard test method for determination of resistance to stable crack extension under low-constraint conditions

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2006-01-01

    1.1 This standard covers the determination of the resistance to stable crack extension in metallic materials in terms of the critical crack-tip-opening angle (CTOAc), ψc and/or the crack-opening displacement (COD), δ5 resistance curve (1). This method applies specifically to fatigue pre-cracked specimens that exhibit low constraint (crack-length-to-thickness and un-cracked ligament-to-thickness ratios greater than or equal to 4) and that are tested under slowly increasing remote applied displacement. The recommended specimens are the compact-tension, C(T), and middle-crack-tension, M(T), specimens. The fracture resistance determined in accordance with this standard is measured as ψc (critical CTOA value) and/or δ5 (critical COD resistance curve) as a function of crack extension. Both fracture resistance parameters are characterized using either a single-specimen or multiple-specimen procedures. These fracture quantities are determined under the opening mode (Mode I) of loading. Influences of environment a...

  12. Standard methods for maintaining adult Apis mellifera in cages under in vitro laboratory conditions

    OpenAIRE

    Williams, Geoffrey R.; Alaux, Cedric; Costa, Cecilia; Csaki, Tamas; Doublet, Vincent; Eisenhardt, Dorothea; Fries, Ingemar; Kuhn, Rolf; McMahon, Dino P.; Medrzycki, Piotr; Murray, Tomas E.; Natsopoulou, Myrsini E.; Neumann, Peter; Oliver, Randy; Paxton, Robert J.

    2013-01-01

    Adult honey bees are maintained in vitro in laboratory cages for a variety of purposes. For example, researchers may wish to perform experiments on honey bees caged individually or in groups to study aspects of parasitology, toxicology, or physiology under highly controlled conditions, or they may cage whole frames to obtain newly emerged workers of known age cohorts. Regardless of purpose, researchers must manage a number of variables, ranging from selection of study subjects (e.g. honey bee...

  13. Standard methods for maintaining adult Apis mellifera in cages under in vitro laboratory conditions

    OpenAIRE

    Williams, G.R.; Alaux, C.; Costa, C.; Csaki, C.; Steen, van der, J.J.M.

    2013-01-01

    Adult honey bees are maintained in vitro in laboratory cages for a variety of purposes. For example, researchers may wish to perform experiments on honey bees caged individually or in groups to study aspects of parasitology, toxicology, or physiology under highly controlled conditions, or they may cage whole frames to obtain freshly emerged workers of known age cohorts. Regardless of purpose, researchers must manage a number of variables, ranging from selection of study subjects (e.g. honey b...

  14. Standard practice for measurement of time-of-wetness on surfaces exposed to wetting conditions as in atmospheric corrosion testing

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    1989-01-01

    1.1 This practice covers a technique for monitoring time-of-wetness (TOW) on surfaces exposed to cyclic atmospheric conditions which produce depositions of moisture. 1.2 The practice is also applicable for detecting and monitoring condensation within a wall or roof assembly and in test apparatus. 1.3 Exposure site calibration or characterization can be significantly enhanced if TOW is measured for comparison with other sites, particularly if this data is used in conjunction with other site-specific instrumentation techniques. 1.4 The values stated in SI units are to be regarded as the standard. This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

  15. Standard Practice for Exposure of Cover Materials for Solar Collectors to Natural Weathering Under Conditions Simulating Operational Mode

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    1995-01-01

    1.1 This practice provides a procedure for the exposure of cover materials for flat-plate solar collectors to the natural weather environment at temperatures that are elevated to approximate operating conditions. 1.2 This practice is suitable for exposure of both glass and plastic solar collector cover materials. Provisions are made for exposure of single and double cover assemblies to accommodate the need for exposure of both inner and outer solar collector cover materials. 1.3 This practice does not apply to cover materials for evacuated collectors or photovoltaics. 1.4 The values stated in SI units are to be regarded as the standard. 1.5 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard to establish appropriate safety and health practices and determine the applicability of regulatory limitations prior to use.

  16. Revisiting olfactory classical conditioning of the proboscis extension response in honey bees: a step toward standardized procedures.

    Science.gov (United States)

    Matsumoto, Yukihisa; Menzel, Randolf; Sandoz, Jean-Christophe; Giurfa, Martin

    2012-10-15

    The honey bee Apis mellifera has emerged as a robust and influential model for the study of classical conditioning thanks to the existence of a powerful Pavlovian conditioning protocol, the olfactory conditioning of the proboscis extension response (PER). In 2011, the olfactory PER conditioning protocol celebrated its 50 years since it was first introduced by Kimihisa Takeda in 1961. In this protocol, individually harnessed honey bees are trained to associate an odor with sucrose solution. The resulting olfactory learning is fast and induces robust olfactory memories that have been characterized at the behavioral, neuronal and molecular levels. Despite the success of this protocol for studying the bases of learning and memory at these different levels, innumerable procedural variants have arisen throughout the years, which render comparative analyses of behavioral performances difficult. Moreover, because even slight variations in conditioning procedures may introduce significant differences in acquisition and retention performances, we revisit olfactory PER conditioning and define here a standardized framework for experiments using this behavioral protocol. To this end, we present and discuss all the methodological steps and details necessary for successful implementation of olfactory PER conditioning. Copyright © 2012 Elsevier B.V. All rights reserved.

  17. Effectiveness of the GAEC cross-compliance standard Ploughing in good soil moisture conditions in soil structure protection

    Directory of Open Access Journals (Sweden)

    Maria Teresa Dell'Abate

    2011-08-01

    Full Text Available Researches have been carried out within the framework on the EFFICOND Project, focused at evaluating the effectiveness of the standards of Good Agricultural and Environmental Conditions (GAECs established for Cross Compliance implementation under EC Regulation 1782/2003. In particular the standard 3.1b deals with soil structure protection through appropriate machinery use, with particular reference to ploughing in good soil moisture conditions. The study deals with the evaluation of soil structure after tillage in tilth and no-tilth conditions at soil moisture contents other than the optimum water content for tillage. The Mean Weight Diameter (MWD of water stable aggregates was used as an indicator of tillage effectiveness. The study was carried out in the period 2008-2009 at six experimental farms belonging to Research Centres and Units of the Italian Agricultural Research Council (CRA with different pedo-climatic and cropping conditions. Farm management and data collection in the different sites were carried out by the local CRA researchers and technicians. The comparison of MWD values in tilth and no tilth theses showed statistically significant differences in most cases, depending on topsoil texture. On clay, clay loam, silty clay, and silty clay loam topsoils a general and significant increase of MWD values under no tilth conditions were observed. No significant differences were observed in silt loam and sandy loam textures, probably due to the weak soil structure of the topsoils. Moreover, ploughing in good soil moisture condition determined higher crop production and less weed development than ploughing in high soil moisture conditions.

  18. Development of a set of multiplex standard polymerase chain reaction assays for the identification of infectious agents from aborted bovine clinical samples.

    Science.gov (United States)

    Tramuta, Clara; Lacerenza, Daniela; Zoppi, Simona; Goria, Mariella; Dondo, Alessandro; Ferroglio, Ezio; Nebbia, Patrizia; Rosati, Sergio

    2011-07-01

    The current study describes the development of a set of 5 multiplex polymerase chain reaction (mPCR) assays for the simultaneous detection of abortive infection agents in bovine fetal tissues, including Brucella spp., Leptospira spp., and Campylobacter fetus (mPCR1); Hammondia heydorni, Neospora caninum, and Toxoplasma gondii (mPCR2); Coxiella burnetii and Chlamydophila psittaci (mPCR3); Mycoplasma bovis, Mycoplasma bovigenitalium, and Ureaplasma diversum (mPCR4); and Bovine viral diarrhea virus (BVDV) and Bovine herpesvirus-1 (BoHV-1; mPCR5). The protocol was tested on different tissue samples collected from 50 aborted bovine fetuses, and it showed that out of the 50 fetuses, 7 (14%, mPCR2) were PCR-positive for N. caninum, 4 (8%, mPCR5) were PCR-positive for BVDV, and 2 (4%, mPCR4) were PCR-positive for U. diversum. The results obtained by using each multiplex PCR were 100% concordant with those obtained by using the respective PCR assays targeting single genes on the same specimens. Moreover, all multiplex PCR assays on clinical samples were compared with reference methods, obtaining a perfect accordance in all samples and confirming the validity of the set of multiplex PCR assays. The proposed set of multiplex PCR assays is, therefore, suitable for the simultaneous detection of the main infectious agents responsible for bovine abortion.

  19. Environmental release of engineered nanomaterials from commercial tiles under standardized abrasion conditions.

    Science.gov (United States)

    Bressot, Christophe; Manier, Nicolas; Pagnoux, Cécile; Aguerre-Chariol, Olivier; Morgeneyer, Martin

    2017-01-15

    The study presented here focuses on commercial antibacterial tiles whose emissivity of (nano) particles due to abrasion has yet barely been investigated. The tiles have been characterized regarding their surface properties and composition throughout their chain-of-use, i.e. from their state of commercialization until the experimental end-of-service life. In contrast to plane standard tiles, their surfaces form hilly surfaces. In the depressions, titanium dioxide is found at the surface, thus theoretically protected by the hilly areas against abrasion on the tile's surface. Furthermore, a deposition technique has been put in place by producers allowing for coating the before mentioned commercial tiles with titanium dioxide, thus being similar to those commercially available. It consists in depositing titanium dioxide on the surface, latter one allowing fixing the first. This development allows for better understanding the future options for product formulation and thus improvement with respect to particle release. The tests reveal the aerosolization from commercial antibacterial tiles of micronic and submicronic particles in the inhalable region or particles that can subjected to be released in the environment (tiles was found to be significantly higher compared to the non coated tiles. Copyright © 2016 Elsevier B.V. All rights reserved.

  20. [Requirement of standardizing anti-HBs assay methods in Japan for HBV infection-preventing strategy--discrepancy of anti-HBs measurements among three different kits widely used in Japan].

    Science.gov (United States)

    Ogata, Norio

    2006-09-01

    The strategy to eliminate hepatitis B virus (HBV) infection by administrating an HB vaccine is changing worldwide; however, this is not the case in Japan. An important concern about the HBV infection-preventing strategy in Japan may be that the assay methods for the antibody to hepatitis B surface antigen (anti-HBs) are not standardized. The minimum protective anti-HBs titer against HBV infection has been established as 10 mIU/ml by World Health Organization (WHO) -standardized assay methods worldwide, but that is still determined as a "positive" test result by the passive hemagglutination (PHA) method in Japan. We compared anti-HBs measurements in given samples among PHA(Mycell II, Institute of Immunology), chemiluminescent enzyme immunoassay (CLEIA) (Lumipulse, Fujirebio), and chemiluminescent immunoassay (CLIA) (Architect, Abbott), all of which are currently in wide use in Japan. First, anti-HBs measurements in serum from individuals who received a yeast-derived recombinant HB vaccine composed of the major surface protein of either subtype adr or subtype ayw were compared. The results clearly showed that in subtype adr-vaccinees CLIA underestimated the anti-HBs amount compared with CLEIA and PHA, but in ayw-vaccinees, the discordance in the measurements among the three kits was not prominent. Second, anti-HBs measurements in standard or calibration solutions of each assay kit were compared. Surprisingly, CLEIA showed higher measurements in all three kit-associated standard or calibration solutions than CLIA. Thus, the anti-HBs titer of 10 mIU/ml is difficult to introduce in Japan as the minimum protective level against HBV infection. Efforts to standardize anti-HBs assay methods are expected to share international evidence about the HBV infection-preventing strategy.

  1. Field Performance versus Standard Test Condition Efficiency of Tandem Solar Cells and the Specific Case of Perovskites/Silicon Devices

    KAUST Repository

    Dupre, Olivier

    2018-01-05

    Multijunction cells may offer a cost-effective route to boost the efficiency of industrial photovoltaics. For any technology to be deployed in the field, its performance under actual operating conditions is extremely important. In this perspective, we evaluate the impact of spectrum, light intensity, and module temperature variations on the efficiency of tandem devices with crystalline silicon bottom cells with a particular focus on perovskite top cells. We consider devices with different efficiencies and calculate their energy yields using field data from Denver. We find that annual losses due to differences between operating conditions and standard test conditions are similar for single-junction and four-terminal tandem devices. The additional loss for the two-terminal tandem configuration caused by current mismatch reduces its performance ratio by only 1.7% when an optimal top cell bandgap is used. Additionally, the unusual bandgap temperature dependence of perovskites is shown to have a positive, compensating effect on current mismatch.

  2. Stability of left ventricular longitudinal and circumferential deformation over time and standard loading conditions.

    Science.gov (United States)

    Kosmala, Wojciech; Przewlocka-Kosmala, Monika; Sharman, James E; Schultz, Martin G; Marwick, Thomas H

    2017-09-01

    Load dependence is an important source of variation in left ventricular (LV) deformation. This impacts on the precision of information obtained from serial measurements. However, it is clinically important to distinguish actual myocardial dysfunction from changes associated with altered loading conditions. We sought to investigate the association of changes of loading parameters with changes in LV longitudinal (GLS) and circumferential (GCS) strains. Baseline and a 12-month follow-up 2D echocardiograms were performed in 191 Stage A heart failure patients with uncomplicated hypertension. These patients underwent simultaneous measurement of conventional and central blood pressures (BPs) and haemodynamic measurements by applanation tonometry. Significant, but weak correlations (r = 0.15-0.28) of LV strain parameters and their changes over the follow-up period were shown for the majority of LV afterload-associated variables, including central and brachial systolic, diastolic, and mean BPs; 24-h systolic and diastolic BPs; peak reservoir and excess pressures; central augmented pressure (CAP) and pulse pressure; augmentation index; and arterial elastance index (EaI). Central mean BP, EaI, and changes in CAP and EaI over follow-up were independent contributors to LV deformation in multivariable analysis. No improvement in the Bland-Altman 95% limits of agreement and correlation coefficients was seen with LV afterload correction of GLS and GCS using central BP indices. LV longitudinal and circumferential strains in a population without apparent heart disease is relatively insusceptible to changes in LV afterload within physiological range, which, therefore, seem unlikely to be a significant confounder in repeated GLS or GCS observations. Published on behalf of the European Society of Cardiology. All rights reserved. © The Author 2016. For permissions please email: journals.permissions@oup.com.

  3. Test of mutagenicity of an irradiated standard diet for laboratory animals in the host-mediated assay with salmonella typhimurium TA 1530

    International Nuclear Information System (INIS)

    Muenzner, R.; Renner, H.W.

    1976-01-01

    Feed irradiated at a dose of 3 Mrad was tested for mutagenic activity in the host-mediated assay with the mouse as host and Salmonella typhimurium TA 1530 as indicator organism. In the in vivo and in the in vitro comparative test the irradiated feed showed no mutagenic effect. (orig.) [de

  4. On the Implementation of the IEC 61850 Standard: Will Different Manufacturer Devices Behave Similarly under Identical Conditions?

    Directory of Open Access Journals (Sweden)

    Mohamad El Hariri

    2016-12-01

    Full Text Available Standardization in smart grid communications is necessary to facilitate complex operations of modern power system functions. However, the strong coupling between the cyber and physical domains of the contemporary grid exposes the system to vulnerabilities and thus places more burden on standards’ developers. As such, standards need to be continuously assessed for reliability and are expected to be implemented properly on field devices. However, the actual implementation of common standards varies between vendors, which may lead to different behaviors of the devices even if present under similar conditions. The work in this paper tested the implementation of the International Electro-technical Commission’s Generic Object Oriented Substation Event GOOSE (IEC 61850 GOOSE messaging protocol on commercial Intelligent Electronic Devices (IEDs and the open source libiec61850 library—also used in commercial devices—which showed different behaviors in identical situations. Based on the test results and analysis of some features of the IEC 61850 GOOSE protocol itself, this paper proposes guidelines and recommendations for proper implementation of the standard functionalities.

  5. A Two-Sinker Densimeter for Accurate Measurements of the Density of Natural Gases at Standard Conditions

    Science.gov (United States)

    Richter, Markus; Kleinrahm, Reiner; Glos, Stefan; Wagner, Wolfgang; Span, Roland; Schley, Peter; Uhrig, Martin

    2010-05-01

    A special reference densimeter has been developed for accurate measurements of densities of natural gases and multicomponent gas mixtures at standard conditions of temperature and pressure ( T s = 273.15 K and p s = 0.101325 MPa). The densimeter covers the range from 0.7 kg · m-3 to 1.3 kg · m-3; the total measurement uncertainty in density is 0.020 % (95 % level of confidence). The measurement principle used is the two-sinker method, which is based on the Archimedes buoyancy principle. The certified calibration laboratory of E.ON Ruhrgas AG, Germany, uses this densimeter to verify the standard densities of certified calibration gases (binary and multicomponent gas mixtures). Moreover, the densimeter is used to determine the compositions of commercially available binary gas mixtures with a small uncertainty of (0.01-0.03) mol%.

  6. ARE METHODS USED TO INTEGRATE STANDARDIZED MANAGEMENT SYSTEMS A CONDITIONING FACTOR OF THE LEVEL OF INTEGRATION? AN EMPIRICAL STUDY

    Directory of Open Access Journals (Sweden)

    Merce Bernardo

    2011-09-01

    Full Text Available Organizations are increasingly implementing multiple Management System Standards (M SSs and considering managing the related Management Systems (MSs as a single system.The aim of this paper is to analyze if methods us ed to integrate standardized MSs condition the level of integration of those MSs. A descriptive methodology has been applied to 343 Spanish organizations registered to, at least, ISO 9001 and ISO 14001. Seven groups of these organizations using different combinations of methods have been analyzed Results show that these organizations have a high level of integration of their MSs. The most common method used, was the process map. Organizations using a combination of different methods achieve higher levels of integration than those using a single method. However, no evidence has been found to confirm the relationship between the method used and the integration level achieved.

  7. Assessing behind armor blunt trauma (BABT) under NIJ standard-0101.04 conditions using human torso models.

    Science.gov (United States)

    Merkle, Andrew C; Ward, Emily E; O'Connor, James V; Roberts, Jack C

    2008-06-01

    Although soft armor vests serve to prevent penetrating wounds and dissipate impact energy, the potential of nonpenetrating injury to the thorax, termed behind armor blunt trauma, does exist. Currently, the ballistic resistance of personal body armor is determined by impacting a soft armor vest over a clay backing and measuring the resulting clay deformation as specified in National Institute of Justice (NIJ) Standard-0101.04. This research effort evaluated the efficacy of a physical Human Surrogate Torso Model (HSTM) as a device for determining thoracic response when exposed to impact conditions specified in the NIJ Standard. The HSTM was subjected to a series of ballistic impacts over the sternum and stomach. The pressure waves propagating through the torso were measured with sensors installed in the organs. A previously developed Human Torso Finite Element Model (HTFEM) was used to analyze the amount of tissue displacement during impact and compared with the amount of clay deformation predicted by a validated finite element model. All experiments and simulations were conducted at NIJ Standard test conditions. When normalized by the response at the lowest threat level (Level I), the clay deformations for the higher levels are relatively constant and range from 2.3 to 2.7 times that of the base threat level. However, the pressures in the HSTM increase with each test level and range from three to seven times greater than Level I depending on the organ. The results demonstrate the abilities of the HSTM to discriminate between threat levels, impact conditions, and impact locations. The HTFEM and HSTM are capable of realizing pressure and displacement differences because of the level of protection, surrounding tissue, and proximity to the impact point. The results of this research provide insight into the transfer of energy and pressure wave propagation during ballistic impacts using a physical surrogate and computational model of the human torso.

  8. Standard test method for non-destructive assay of nuclear material in waste by passive and active neutron counting using a differential Die-away system

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2009-01-01

    1.1 This test method covers a system that performs nondestructive assay (NDA) of uranium or plutonium, or both, using the active, differential die-away technique (DDT), and passive neutron coincidence counting. Results from the active and passive measurements are combined to determine the total amount of fissile and spontaneously-fissioning material in drums of scrap or waste. Corrections are made to the measurements for the effects of neutron moderation and absorption, assuming that the effects are averaged over the volume of the drum and that no significant lumps of nuclear material are present. These systems are most widely used to assay low-level and transuranic waste, but may also be used for the measurement of scrap materials. The examples given within this test method are specific to the second-generation Los Alamos National Laboratory (LANL) passive-active neutron assay system. 1.1.1 In the active mode, the system measures fissile isotopes such as 235U and 239Pu. The neutrons from a pulsed, 14-MeV ne...

  9. Controlling variation in the comet assay

    Directory of Open Access Journals (Sweden)

    Andrew Richard Collins

    2014-10-01

    Full Text Available Variability of the comet assay is a serious issue, whether it occurs from experiment to experiment in the same laboratory, or between different laboratories analysing identical samples. Do we have to live with high variability, just because the comet assay is a biological assay rather than analytical chemistry? Numerous attempts have been made to limit variability by standardising the assay protocol, and the critical steps in the assay have been identified; agarose concentration, duration of alkaline incubation, and electrophoresis conditions (time, temperature and voltage gradient are particularly important. Even when these are controlled, variation seems to be inevitable. It is helpful to include in experiments reference standards, i.e. cells with a known amount of specific damage to the DNA. They can be aliquots frozen from a single large batch of cells, either untreated (negative controls or treated with, for example, H2O2 or X-rays to induce strand breaks (positive control for the basic assay, or photosensitiser plus light to oxidise guanine (positive control for Fpg- or OGG1-sensitive sites. Reference standards are especially valuable when performing a series of experiments over a long period - for example, analysing samples of white blood cells from a large human biomonitoring trial - to check that the assay is performing consistently, and to identify anomalous results necessitating a repeat experiment. The reference values of tail intensity can also be used to iron out small variations occurring from day to day. We present examples of the use of reference standards in human trials, both within one laboratory and between different laboratories, and describe procedures that can be used to control variation.

  10. Protein folding: Defining a standard set of experimental conditions and a preliminary kinetic data set of two-state proteins

    DEFF Research Database (Denmark)

    Maxwell, Karen L.; Wildes, D.; Zarrine-Afsar, A.

    2005-01-01

    Recent years have seen the publication of both empirical and theoretical relationships predicting the rates with which proteins fold. Our ability to test and refine these relationships has been limited, however, by a variety of difficulties associated with the comparison of folding and unfolding ...... efforts is to set uniform standards for the experimental community and to initiate an accumulating, self-consistent data set that will aid ongoing efforts to understand the folding process....... constructs. The lack of a single approach to data analysis and error estimation, or even of a common set of units and reporting standards, further hinders comparative studies of folding. In an effort to overcome these problems, we define here a consensus set of experimental conditions (25°C at pH 7.0, 50 m...... rates, thermodynamics, and structure across diverse sets of proteins. These difficulties include the wide, potentially confounding range of experimental conditions and methods employed to date and the difficulty of obtaining correct and complete sequence and structural details for the characterized...

  11. THE BECOMING OF INFORMATION CULTURE IN THE CONDITIONS OF THE FEDERAL STATE EDUCATIONAL STANDARD OF VOCATIONAL EDUCATION’S IMPLEMENTATION

    Directory of Open Access Journals (Sweden)

    Lapina Svetlana Nikolaevna

    2013-05-01

    Full Text Available This article examines the approaches to the definition of “information culture”, its components, the system of personal values needed to succeed in the information and professional activities, the problem of students’ information culture formation in the modern information society. The analysis of the implementation of the Federal state educational standard of vocational education in "teaching in primary schools" is held. The variable part cycles of the basic professional educational programs is distributed on the base of the local professional community’s research and additional competencies. Such subjects as “Russian language and Speech”, “The cultural world of students”, “Ethics in business communication” are introduced through the variable part of the educational standard. The general amount of hours for such subject as «Computer science, information and communication technology in the professional activity" is increased. The results of the special study reveal the level of information culture of the future primary school teachers. According to the results it can be concluded that insufficient level of information culture’s development is impossible for a successful career and self-fulfillment in the present conditions. The article proposes the directions for the formation of future primary school teachers’ information culture in the implementation of the federal state educational standard of vocational education. According to the results of this research it is possible to tell about the effectiveness of these directions’ implementation.

  12. THE BECOMING OF INFORMATION CULTURE IN THE CONDITIONS OF THE FEDERAL STATE EDUCATIONAL STANDARD OF VOCATIONAL EDUCATION’S IMPLEMENTATION

    Directory of Open Access Journals (Sweden)

    Светлана Николаевна Лапина

    2013-07-01

    Full Text Available This article examines the approaches to the definition of “information culture”, its components, the system of personal values needed to succeed in the information and professional activities, the problem of students’ information culture formation in the modern information society. The analysis of the implementation of the Federal state educational standard of vocational education in "teaching in primary schools" is held. The variable part cycles of the basic professional educational programs is distributed on the base of the local professional community’s research and additional competencies. Such subjects as “Russian language and Speech”, “The cultural world of students”, “Ethics in business communication” are introduced through the variable part of the educational standard. The general amount of hours for such subject as «Computer science, information and communication technology in the professional activity" is increased. The results of the special study reveal the level of information culture of the future primary school teachers. According to the results it can be concluded that insufficient level of information culture’s development is impossible for a successful career and self-fulfillment in the present conditions. The article proposes the directions for the formation of future primary school teachers’ information culture in the implementation of the federal state educational standard of vocational education. According to the results of this research it is possible to tell about the effectiveness of these directions’ implementation.DOI: http://dx.doi.org/10.12731/2218-7405-2013-5-31

  13. conditions

    Directory of Open Access Journals (Sweden)

    M. Venkatesulu

    1996-01-01

    Full Text Available Solutions of initial value problems associated with a pair of ordinary differential systems (L1,L2 defined on two adjacent intervals I1 and I2 and satisfying certain interface-spatial conditions at the common end (interface point are studied.

  14. Growth curves and the international standard: How children's growth reflects challenging conditions in rural Timor-Leste.

    Science.gov (United States)

    Spencer, Phoebe R; Sanders, Katherine A; Judge, Debra S

    2018-02-01

    Population-specific growth references are important in understanding local growth variation, especially in developing countries where child growth is poor and the need for effective health interventions is high. In this article, we use mixed longitudinal data to calculate the first growth curves for rural East Timorese children to identify where, during development, deviation from the international standards occurs. Over an eight-year period, 1,245 children from two ecologically distinct rural areas of Timor-Leste were measured a total of 4,904 times. We compared growth to the World Health Organization (WHO) standards using z-scores, and modeled height and weight velocity using the SuperImposition by Translation And Rotation (SITAR) method. Using the Generalized Additive Model for Location, Scale and Shape (GAMLSS) method, we created the first growth curves for rural Timorese children for height, weight and body mass index (BMI). Relative to the WHO standards, children show early-life growth faltering, and stunting throughout childhood and adolescence. The median height and weight for this population tracks below the WHO fifth centile. Males have poorer growth than females in both z-BMI (p = .001) and z-height-for-age (p = .018) and, unlike females, continue to grow into adulthood. This is the most comprehensive investigation to date of rural Timorese children's growth, and the growth curves created may potentially be used to identify future secular trends in growth as the country develops. We show significant deviation from the international standard that becomes most pronounced at adolescence, similar to the growth of other Asian populations. Males and females show different growth responses to challenging conditions in this population. © 2017 Wiley Periodicals, Inc.

  15. Standards of Conditions During Preparations for the Summer Paralympic Games Between 2004 and 2012 Assessed by Polish Athletes

    Directory of Open Access Journals (Sweden)

    Sobiecka Joanna

    2015-12-01

    Full Text Available The quality of training conditions affects sporting success, injuries and health. The aim of the work was to present the conditions during the preparations of Polish athletes for the Summer Paralympic Games 2004-2012. The study encompassed 271 paralympians: Athens (91, Beijing (89 and London (91, competing in 13 disciplines. The research was based on a two-part questionnaire by Kłodecka-Różalska adjusted for disabled sports, and was conducted one month before each PG. Part 1 contained 20 closed-ended questions regarding conditions during preparations, while Part 2 concerned socio-demographic and sports-related data. Three levels of conditions: good, satisfactory and poor, were identified. The analysis showed that while the relationships between the athletes were good in all the preparatory periods, the co-operation with the paralympic coaches worsened. The standards of accommodation, food and sports facilities lowered. Personal orthopaedic supply was satisfactory in London; personal sporting equipment was good at all PG. The quality of medical care was the highest in London. The co-operation with physicians, physiotherapists and massage therapists was satisfactory. Consultations with the dietician were sporadic and assessed as poor. Psychological consultations were rare but satisfactory in Beijing and London. Contacts with the mass media were poor at all PG. Although combining private life, work, and education with sport was satisfactory, it was increasingly difficult to manage, particularly before London. The conditions during preparations for the PG 2004-2012 varied. Improvement was noticed only in the quality of medical care and personal orthopaedic supply.

  16. Standards of Conditions During Preparations for the Summer Paralympic Games Between 2004 and 2012 Assessed by Polish Athletes.

    Science.gov (United States)

    Sobiecka, Joanna; Gawroński, Wojciech; Kądziołka, Marta; Kruszelnicki, Paweł; Kłodecka-Różalska, Jadwiga; Plinta, Ryszard

    2015-11-22

    The quality of training conditions affects sporting success, injuries and health. The aim of the work was to present the conditions during the preparations of Polish athletes for the Summer Paralympic Games 2004-2012. The study encompassed 271 paralympians: Athens (91), Beijing (89) and London (91), competing in 13 disciplines. The research was based on a two-part questionnaire by Kłodecka-Różalska adjusted for disabled sports, and was conducted one month before each PG. Part 1 contained 20 closed-ended questions regarding conditions during preparations, while Part 2 concerned socio-demographic and sports-related data. Three levels of conditions: good, satisfactory and poor, were identified. The analysis showed that while the relationships between the athletes were good in all the preparatory periods, the co-operation with the paralympic coaches worsened. The standards of accommodation, food and sports facilities lowered. Personal orthopaedic supply was satisfactory in London; personal sporting equipment was good at all PG. The quality of medical care was the highest in London. The co-operation with physicians, physiotherapists and massage therapists was satisfactory. Consultations with the dietician were sporadic and assessed as poor. Psychological consultations were rare but satisfactory in Beijing and London. Contacts with the mass media were poor at all PG. Although combining private life, work, and education with sport was satisfactory, it was increasingly difficult to manage, particularly before London. The conditions during preparations for the PG 2004-2012 varied. Improvement was noticed only in the quality of medical care and personal orthopaedic supply.

  17. Hyaluronic Acid Assays

    DEFF Research Database (Denmark)

    Itenov, Theis Skovsgaard; Kirkby, Nikolai S; Bestle, Morten H

    2016-01-01

    BACKGROUD: Hyaluronic acid (HA) is proposed as a marker of functional liver capacity. The aim of the present study was to compare a new turbidimetric assay for measuring HA with the current standard method. METHODS: HA was measured by a particle-enhanced turbidimetric immunoassay (PETIA) and enzyme...

  18. FLUIDICS DEVICE FOR ASSAY

    DEFF Research Database (Denmark)

    2007-01-01

    The present invention relates to a device for use in performing assays on standard laboratory solid supports whereon chemical entities are attached. The invention furthermore relates to the use of such a device and a kit comprising such a device. The device according to the present invention is a...

  19. A standardized approach for estimating the permeability of plastic films to soil fumigants under various field and environmental conditions.

    Science.gov (United States)

    Papiernik, Sharon K; Yates, Scott R; Chellemi, Daniel O

    2011-01-01

    Minimizing atmospheric emissions of soil fumigants is critical for protecting human and environmental health. Covering the soil surface with a plastic tarp is a common approach to restrict fumigant emissions. The mass transfer of the fumigant vapors through the tarp is often the rate-limiting factor in fumigant emissions. An approach for standardizing measurements of film permeability is proposed that is based on determining the resistance (R) of films to diffusion of fumigants. Using this approach, values were determined for more than 200 film-chemical combinations under a range of temperature, relative humidity, and film handling conditions. Resistance to diffusion was specific for each fumigant/film combination, with the largest range of values observed for the fumigant chloropicrin. For each fumigant, decreased with increasing temperature. Changes in film permeability due to increases in temperature or field installation were generally less than a factor of five. For one film, values determined under conditions of very high relative humidity (approximately 100%) were at least 100 times lower than when humidity was very low (approximately 2%). This approach simplifies the selection of appropriate films for soil fumigation by providing rapid, reproducible, and precise measurements of their permeability to specific fumigants and application conditions. by the American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America, Inc.

  20. Standard test method for nondestructive assay of nuclear material in scrap and waste by passive-Active neutron counting using 252Cf shuffler

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2008-01-01

    1.1 This test method covers the nondestructive assay of scrap and waste items for U, Pu, or both, using a 252Cf shuffler. Shuffler measurements have been applied to a variety of matrix materials in containers of up to several 100 L. Corrections are made for the effects of matrix material. Applications of this test method include measurements for safeguards, accountability, TRU, and U waste segregation, disposal, and process control purposes (1, 2, 3). 1.1.1 This test method uses passive neutron coincidence counting (4) to measure the 240Pu-effective mass. It has been used to assay items with total Pu contents between 0.03 g and 1000 g. It could be used to measure other spontaneously fissioning isotopes such as Cm and Cf. It specifically describes the approach used with shift register electronics; however, it can be adapted to other electronics. 1.1.2 This test method uses neutron irradiation with a moveable Cf source and counting of the delayed neutrons from the induced fissions to measure the 235U equiva...

  1. Hormone assay

    International Nuclear Information System (INIS)

    Eisentraut, A.M.

    1977-01-01

    An improved radioimmunoassay is described for measuring total triiodothyronine or total thyroxine levels in a sample of serum containing free endogenous thyroid hormone and endogenous thyroid hormone bound to thyroid hormone binding protein. The thyroid hormone is released from the protein by adding hydrochloric acid to the serum. The pH of the separated thyroid hormone and thyroid hormone binding protein is raised in the absence of a blocking agent without interference from the endogenous protein. 125 I-labelled thyroid hormone and thyroid hormone antibodies are added to the mixture, allowing the labelled and unlabelled thyroid hormone and the thyroid hormone antibody to bind competitively. This results in free thyroid hormone being separated from antibody bound thyroid hormone and thus the unknown quantity of thyroid hormone may be determined. A thyroid hormone test assay kit is described for this radioimmunoassay. It provides a 'single tube' assay which does not require blocking agents for endogenous protein interference nor an external solid phase sorption step for the separation of bound and free hormone after the competitive binding step; it also requires a minimum number of manipulative steps. Examples of the assay are given to illustrate the reproducibility, linearity and specificity of the assay. (UK)

  2. Impact of Assay conditions on activity estimate and kinetics comparison of Aspergillus niger PhyA and Escherichia coli AppA2 phytases

    Science.gov (United States)

    This study was to compare three phytase activity assays and kinetics of Aspergillus niger PhyA and Escherichia coli AppA2 phytases expressed in Pichia pastoris at the observed stomach pH of 3.5. In Experiment 1, equivalent phytase activities in the crude preparations of PhyA and AppA2 were tested ...

  3. The Disposition of Water Supply and Demand in Cameroon: What Potential for what Standard of Living Conditions?

    Directory of Open Access Journals (Sweden)

    Oumar Saidou Baba

    2017-04-01

    Full Text Available Aim/purpose - This paper attempts to appraise the potential of water resources for Cameroon and the standard of living conditions confronting people in the country. Design/methodology/approach - A simple descriptive method of data analysis is adopted using analytical tools such as percentages, tables, and means to achieve the objectives of the inquiry. Data for the study were generated from personal observations in one hand and collected from water resources literature, on the other hand. Findings - With the help of the data gathered, the paper establishes that despite the existence of abundant water resources in Cameroon the standard of living conditions of people with respect to basic needs of survival such as drinking water, improved sanitation services, and electricity supply is far below expectation. Research implications/limitations - The main implication of the study is that in spite of the surplus volume of water resources (325.96 km3 or 95.12% of annual total water resources endowment in Cameroon, the population benefits marginally from it due to the mismanagement of resources and misplacement of priorities as obtained in most sub-Saharan African countries. One limitation of this study is that the use of limited primary data in the investigation offers no room toward establishing the extent of water resources allocation to the various users of water in the country. Originality/value/contribution - The paper suggests that the government of Cameroon should encourage the population to run community basic social services projects and subsidize the activities of such ventures in kind through technical assistance or in cash.

  4. Dynamic behavior structural response and capacity evaluation of the standardized WWER-1000 nuclear power plants subjected to severe loading conditions

    International Nuclear Information System (INIS)

    Ambriashvili, Y.K.; Krutzik, N.J.

    1993-01-01

    In order to verify the structural capacity of standardized WWER-1000 MW nuclear power plants, comprehensive static and dynamic analyses were performed in cooperation between Siemens and Atomenergoprojekt. The main goal of these investigations was to perform of a number of seismic analyses of standardized WWER-1000 reactor buildings on the basis of 13 given seismological inputs, taking into account the local soil conditions at 17 different sites defined by in-situ investigations. The analyses were based on appropriate mathematical models (equivalent beam models as well as detailed spatial surface element models) of the coupled vibrating structures (base structure, outer structure, containment, inner structure) and of the layered soil. The analyses were mainly performed using the indirect method (substructure method). Based on the results of the seismic analysis as well as the results of static analysis (pressure and temperature due to LOCA, dead weight, prestressing) an assessment was made of the seismic safety of the containment and the reactor building. Using a complex 3-dimensional model of the structure and the soil, the influence of the flexibility of the basement structure on the structural response was also studied. The structural analyses of the WWER-1000 reactor building led to the conclusion that its design accounts well for the main factors governing the dynamic behavior of the building. The assessment of the forces acting in the structures shows that the bearing capacity of the analyzed building structure corresponds to an earthquake intensity of about 0.2 g to 0.25 g

  5. Standard Practice for Exposure of Solar Collector Cover Materials to Natural Weathering Under Conditions Simulating Stagnation Mode

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    1992-01-01

    1.1 This practice covers a procedure for the exposure of solar collector cover materials to the natural weather environment at elevated temperatures that approximate stagnation conditions in solar collectors having a combined back and edge loss coefficient of less than 1.5 W/(m2 · °C). 1.2 This practice is suitable for exposure of both glass and plastic solar collector cover materials. Provisions are made for exposure of single and double cover assemblies to accommodate the need for exposure of both inner and outer solar collector cover materials. 1.3 This practice does not apply to cover materials for evacuated collectors, photovoltaic cells, flat-plate collectors having a combined back and edge loss coefficient greater than 1.5 W/(m2 ·° C), or flat-plate collectors whose design incorporates means for limiting temperatures during stagnation. 1.4 This standard does not purport to address all of the safety concerns, if any, associated with its use. It is the responsibility of the user of this standard t...

  6. TRAK App Suite: A Web-Based Intervention for Delivering Standard Care for the Rehabilitation of Knee Conditions.

    Science.gov (United States)

    Spasić, Irena; Button, Kate; Divoli, Anna; Gupta, Satyam; Pataky, Tamas; Pizzocaro, Diego; Preece, Alun; van Deursen, Robert; Wilson, Chris

    2015-10-16

    Standard care for the rehabilitation of knee conditions involves exercise programs and information provision. Current methods of rehabilitation delivery struggle to keep up with large volumes of patients and the length of treatment required to maximize the recovery. Therefore, the development of novel interventions to support self-management is strongly recommended. Such interventions need to include information provision, goal setting, monitoring, feedback, and support groups, but the most effective methods of their delivery are poorly understood. The Internet provides a medium for intervention delivery with considerable potential for meeting these needs. The objective of this study was to demonstrate the feasibility of a Web-based app and to conduct a preliminary review of its practicability as part of a complex medical intervention in the rehabilitation of knee disorders. This paper describes the development, implementation, and usability of such an app. An interdisciplinary team of health care professionals and researchers, computer scientists, and app developers developed the TRAK app suite. The key functionality of the app includes information provision, a three-step exercise program based on a standard care for the rehabilitation of knee conditions, self-monitoring with visual feedback, and a virtual support group. There were two types of stakeholders (patients and physiotherapists) that were recruited for the usability study. The usability questionnaire was used to collect both qualitative and quantitative information on computer and Internet usage, task completion, and subjective user preferences. A total of 16 patients and 15 physiotherapists participated in the usability study. Based on the System Usability Scale, the TRAK app has higher perceived usability than 70% of systems. Both patients and physiotherapists agreed that the given Web-based approach would facilitate communication, provide information, help recall information, improve understanding

  7. Standard test method for nondestructive assay of special nuclear material in low density scrap and waste by segmented passive gamma-Ray scanning

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2010-01-01

    1.1 This test method covers the transmission-corrected nondestructive assay (NDA) of gamma-ray emitting special nuclear materials (SNMs), most commonly 235U, 239Pu, and 241Am, in low-density scrap or waste, packaged in cylindrical containers. The method can also be applied to NDA of other gamma-emitting nuclides including fission products. High-resolution gamma-ray spectroscopy is used to detect and measure the nuclides of interest and to measure and correct for gamma-ray attenuation in a series of horizontal segments (collimated gamma detector views) of the container. Corrections are also made for counting losses occasioned by signal processing limitations (1-3). 1.2 There are currently several systems in use or under development for determining the attenuation corrections for NDA of radioisotopic materials (4-8). A related technique, tomographic gamma-ray scanning (TGS), is not included in this test method (9, 10, 11). 1.2.1 This test method will cover two implementations of the Segmented Gamma Scanning ...

  8. 40 CFR 80.527 - Under what conditions may motor vehicle diesel fuel subject to the 15 ppm sulfur standard be...

    Science.gov (United States)

    2010-07-01

    ... vehicle diesel fuel subject to the 15 ppm sulfur standard be downgraded to motor vehicle diesel fuel... Diesel Fuel; Nonroad, Locomotive, and Marine Diesel Fuel; and ECA Marine Fuel Motor Vehicle Diesel Fuel Standards and Requirements § 80.527 Under what conditions may motor vehicle diesel fuel subject to the 15...

  9. Evaluation of the Ross fast solution of Richards' equation in unfavourable conditions for standard finite element methods

    International Nuclear Information System (INIS)

    Crevoisier, D.; Voltz, M.; Chanzy, A.

    2009-01-01

    Ross [Ross PJ. Modeling soil water and solute transport - fast, simplified numerical solutions. Agron J 2003;95:1352-61] developed a fast, simplified method for solving Richards' equation. This non-iterative 1D approach, using Brooks and Corey [Brooks RH, Corey AT. Hydraulic properties of porous media. Hydrol. papers, Colorado St. Univ., Fort Collins: 1964] hydraulic functions, allows a significant reduction in computing time while maintaining the accuracy of the results. The first aim of this work is to confirm these results in a more extensive set of problems, including those that would lead to serious numerical difficulties for the standard numerical method. The second aim is to validate a generalisation of the Ross method to other mathematical representations of hydraulic functions. The Ross method is compared with the standard finite element model, Hydrus-1D [Simunek J, Sejna M, Van Genuchten MTh. The HYDRUS-1D and HYDRUS-2D codes for estimating unsaturated soil hydraulic and solutes transport parameters. Agron Abstr 357; 1999]. Computing time, accuracy of results and robustness of numerical schemes are monitored in 1D simulations involving different types of homogeneous soils, grids and hydrological conditions. The Ross method associated with modified Van Genuchten hydraulic functions [Vogel T, Cislerova M. On the reliability of unsaturated hydraulic conductivity calculated from the moisture retention curve. Transport Porous Media 1988:3:1-15] proves in every tested scenario to be more robust numerically, and the compromise of computing time/accuracy is seen to be particularly improved on coarse grids. Ross method run from 1.25 to 14 times faster than Hydrus-1D. (authors)

  10. Rover waste assay system

    International Nuclear Information System (INIS)

    Akers, D.W.; Stoots, C.M.; Kraft, N.C.; Marts, D.J.

    1997-01-01

    The Rover Waste Assay System (RWAS) is a nondestructive assay system designed for the rapid assay of highly-enriched 235 U contaminated piping, tank sections, and debris from the Rover nuclear rocket fuel processing facility at the Idaho Chemical Processing Plant. A scanning system translates a NaI(Tl) detector/collimator system over the structural components where both relative and calibrated measurements for 137 Cs are made. Uranium-235 concentrations are in operation and is sufficiently automated that most functions are performed by the computer system. These functions include system calibration, problem identification, collimator control, data analysis, and reporting. Calibration of the system was done through a combination of measurements on calibration standards and benchmarked modeling. A description of the system is presented along with the methods and uncertainties associated with the calibration and analysis of the system for components from the Rover facility. 4 refs., 2 figs., 4 tabs

  11. Detection of Salmonella enterica serovar Enteritidis using real time PCR, immunocapture assay, PNA FISH and standard culture methods in different types of food samples.

    Science.gov (United States)

    Almeida, C; Cerqueira, L; Azevedo, N F; Vieira, M J

    2013-01-15

    Several methods for the rapid and specific detection of Salmonella in food samples have been described. Here, we compare 4 of those methods in terms of assay time, procedure complexity, detection limit, sensitivity, specificity and accuracy. Milk, eggs and mayonnaise samples were artificially contaminated with Salmonella enterica serovar Enteritidis cell concentrations ranging from 1×10(-2) to 1×10(2) CFU per 25 g or ml of food. Samples were then pre-enriched and analyzed by either: i) real-time PCR, using the iQ-Check Salmonella kit; ii) immunocapture, using the RapidChek SELECT Salmonella; iii) a peptide nucleic acid fluorescence in situ hybridization (PNA FISH) method and iv) the traditional bacteriological method ISO 6579:2002. All methods were able to detect Salmonella in the different types of food matrixes and presented a similar detection level of 1CFU per 25 g or ml of food sample. The immunocapture and the PNA FISH methods proved to be very reliable, as their results were 100% in agreement with the ISO method. However, real-time PCR presented a significant number of false positives, which resulted in a specificity of 55.6% (CI 95%, 31.3-77.6) and an accuracy of 82.2% (CI 95%, 63.2-91.4) for this method. Sensitivity was 100% since no false negative results were observed. In conclusion, the implementation of these molecular techniques, mainly the immunocapture and PNA-FISH methods, provides a reliable and less time-consuming alternative for the detection of Salmonella spp. in food samples. Copyright © 2012 Elsevier B.V. All rights reserved.

  12. Standardize or Diversify Experimental Conditions in Ecotoxicology? A Case Study on Herbicide Toxicity to Larvae of Two Anuran Amphibians.

    Science.gov (United States)

    Mikó, Zsanett; Ujszegi, János; Gál, Zoltán; Hettyey, Attila

    2017-11-01

    Despite a steeply increasing number of ecotoxicological studies on the effects of pesticides on nontarget organisms, studies assessing the adequacy and reliability of different experimental approaches have remained scarce. We scrutinized effects of a glyphosate-based herbicide on larvae of two European anuran amphibians by estimating species-specific LC50 values, assessing how an additional stress factor may influence outcomes, and investigating whether replicate experiments yielded qualitatively the same results. We exposed Rana dalmatina and Bufo bufo tadpoles to two predator treatments (no predator vs. predator chemical cues) combined with varying herbicide concentrations, repeated the experiment with a subset of the experimental treatments and partly with slight modifications 1 week later and assessed survival. Our results indicated that the herbicide was moderately toxic to tadpoles. The presence of predator chemical cues did not affect the lethality of the herbicide in either species. The estimated sensitivity of R. dalmatina tadpoles varied considerably across experiments, whereas in case of B. bufo LC50 values remained very similar. Our results suggest that differences in the experimental setup may often have no influence on the measured effects of pesticides, whereas replicated experiments can deliver widely differing results in other cases, perhaps depending on the studied species, the population origin of the tested individuals, or the test conditions. This draws attention to the suggestion that strict standardization may not deliver widely applicable insights into the toxicity of contaminants and, instead, intentionally introducing variation into the design of ecotoxicological experiments and replicating entire experiments may prove highly beneficial.

  13. Zoobenthic Indicators of Environmental Condition At Great Lakes Coastal Margins Derived From Standardized Multivariate Analyses Across Anthropogenic Stress Gradients.

    Science.gov (United States)

    Ciborowski, J. J.; Johnson, L. B.; Gathman, J. P.; Brady, V. J.; Holland, J.; Hollenhorst, T.; Schuldt, J. A.; Host, G. E.; Richards, C.

    2005-05-01

    We used multivariate methods to derive and map multiple indices of zoobenthic community composition across anthropogenic stress gradients at Great Lakes coastal margins (shorelines - 10 m depth). We collected up to 24 benthic samples and measured environmental characteristics in each of 150 "segment sheds", ranging from minimally disturbed (=reference) to greatly modified. "K" distinct suites of co-occurring biota from minimally stressed locations were identified using cluster analysis. Discriminant function analysis identified the environmental attributes of sample locations best distinguishing each of the suites (=habitat types). Benthic community data of all samples from each habitat type (ranging across the entire stress gradient) were individually ordinated. The benthic taxa best characterizing the ends of each stress gradient were used to create a gradient-specific biotic index with a standardized scale (0=minimum, 100=maximum stress) for each habitat type, providing a stress-specific score for each sample. Contour surface analysis of mapped sample-specific scores shows the spatial pattern and limits of the overall biotic response to stress, independently of habitat type. The spatial concordance between biotic index scores and environmental stress can indicate the degree to which equivalent-to-reference conditions exist in a particular area.

  14. Endoproteolytic activity assay in malting barley

    Directory of Open Access Journals (Sweden)

    Blanca Gómez Guerrero

    2013-12-01

    Full Text Available Hydrolysis of barley proteins into peptides and amino acids is one of the most important processes during barley germination.The degradation of the endosperm stored proteins facilitates water and enzyme movements, enhances modification, liberates starch granules and increases soluble amino nitrogen. Protease activity is the result of the activities of a mixture of exo- and endo-proteases. The barley proteins are initially solubilized by endo-proteases and the further by exo-proteases. Four classes of endo-proteases have been described: serine-proteases, cysteine-proteases, aspartic-proteases and metallo-proteases. The objective of this work was to develop a rapid and colorimetric enzymatic assay to determine the endo-proteolytic activity of the four endo-protease classes using two different substrates: azo-gelatin and azo-casein. Optimum conditions for the assays such as: pH,reaction time and temperature and absorbance scale were determined. Azo-gelatin presented several difficulties in standardizing an “in solution” assay. On the other hand, azo-casein allowed standardization of the assay for the four enzyme classes to produce consistent results. The endo-proteoteolytic method developed was applied to determine the endo-protease activity in barley, malt and wort.

  15. Development of 23 individual TaqMan® real-time PCR assays for identifying common foodborne pathogens using a single set of amplification conditions.

    Science.gov (United States)

    Cremonesi, Paola; Pisani, Laura Francesca; Lecchi, Cristina; Ceciliani, Fabrizio; Martino, Pieranna; Bonastre, Armand Sanchez; Karus, Avo; Balzaretti, Claudia; Castiglioni, Bianca

    2014-10-01

    Most of the acute intestinal diseases are caused by foodborne pathogens with infants and elderly people being at major risk. The aim of this study was to develop a procedure to simultaneously detect 20 foodborne pathogens in complex alimentary matrices such as milk, cheese and meat. The list of targets include, among the others, Listeria spp., Salmonella spp., Shigella spp., Escherichia coli spp., Campylobacter spp., Clostridium spp. and Staphylococcus aureus. The accuracy of detection was determined by using ATCC strains as positive and negative controls. The achieved sensitivity of each of assays was 1 pg of genomic DNA, which was equivalent to ∼1 cfu. The working ranges of the TaqMan(®) Real-time PCR assays, when used quantitatively on cheese and meat samples inoculated with serial dilution of Listeria spp., Listeria monocytogenes, S. aureus, Salmonella enterica, Shigella boydii, E. coli O157:H7, Bacillus cereus, Campylobacter coli, Yersinia enterocolitica, Enterobacter sakazakii and Pseudomonas aeruginosa was 10(8) cfu/g to 10(4) cfu/g. No matrix interferences were observed. Copyright © 2014 Elsevier Ltd. All rights reserved.

  16. Assessment of Humidity Conditions and Trends Based on Standardized Precipitation Evapotranspiration Index (SEPI Over Different Climatic Regions of Iran

    Directory of Open Access Journals (Sweden)

    Mohammad Ghabaei S

    2017-01-01

    Full Text Available Introduction: Drought is a recurrent feature of climate that caused by deficiency of precipitation over time. Due to the rise in water demand and alarming climate change, recent year’s observer much focus on drought and drought conditions. A multiple types of deficits and relevant temporal scales can be achieved through the construction of a joint indicator that draws on information from multiple sources and will therefore enable better assessment of drought characteristics including return period, persistent and severity. The Standardized Precipitation Evapotranspiration Index (SPEI combines information from precipitation and temperature in the form of water surplus or deficit according to Standardized Precipitation Index (SPI. Rainfall over some regions of Iran during some resent year was below average while mean and maximum temperatures were very high during this period, as was evaporation. This would suggest that drought conditions were worse than in previous recent periods with similarly low rainfall. The main objective of this study is to assess the influences of humidity on the SPEI index and investigate its relation with SPI and Reconnaissance Drought Index (RDI over six different climatic regions in Iran. Materials and Methods: Iran has different climatic conditions which vary from desert in central part to costal wet near the Caspian Sea. In this study the selection of stations was done based on Alijani et al (2008 climatic classification. We chose 11 synoptic stations from six different climatic classes including costal wet (Rasht and Babolsar, semi mountains (Mashhad and Tabriz, mountains (Shiraz and Khoram Abad, semi-arid (Tehran and Semnan, arid (Kerman and Yazd and costal desert (Bandar Abas. The Meteorological datasets for the aforementioned stations were obtained from the Iran Meteorological Organization (IRIMO for the period 1960-2010. The compiled data included average monthly values of precipitation, minimum and maximum air

  17. PROFESSIONAL TEACHERS’ DEVELOPMENT OF EDUCATIONAL COMPLEXES IN THE CONDITIONS OF INTRODUCTION OF THE FEDERAL STATE EDUCATIONAL STANDARD

    Directory of Open Access Journals (Sweden)

    Elena A. Sidenko

    2016-01-01

    Full Text Available RETRACTED ARTICLEThe aim of this publication is to familiarize readers with the particularities of training of school teams for the introduction of the Federal State Educational Standard (hereinafter – FSES in the initial stage and considering the features of creating a component of model, namely, program of management training to the introduction of the (FSES. The purpose of the study is to developmethodological bases of construction models of implementation of the Federal state educational standard as a system innovation.Methods. The theoretical and methodological basis of the investigation involves: concepts of motivation of work and reflexive-humanistic psychology; ideas of contextual and projective training, and also acmeological approach to continuous education of adults.Results and scientific novelty. The specifics of the program of professional development of shots for education corresponding to modern realities are described. This program assumes preparation of the school teams consisting of heads of the educational organizations, their deputies, methodologists and teachers. Adequate forms of carrying out internal and correspondence occupations with the organization of productive educational activity of listeners are selected; the complex of tasks for course and intersession occupations is developed. Training of listeners according to the developed program is based as the organizational and pedagogical cascade and cluster model combining resident instruction of the managerial personnel in the organizations of system of professional development with their innovative activity in the educational organizations. The course includes some levels: the basic, technological and organizational-activity-based. The basic level assumes development of the maintenance of FSES and development of innovative potential of listeners; technological level – technologies of conducting group work on development of FSES are mastered

  18. Inter-experiment variation and dependence on culture conditions in assaying the chemosensitivity of human small cell lung cancer cell lines

    DEFF Research Database (Denmark)

    Roed, H; Christensen, I B; Vindeløv, L L

    1987-01-01

    Sensitivity of five human small cell lung cancer cell lines to doxorubicin was assessed by a double layer agar technique using two different bottom-layers. Neither of the bottom-layers provided proportionality between numbers of cells plated and numbers of colonies, but they were correlated...... by a logarithmic function. Even after correction for lack of proportionality the two assay systems provided significantly different dose-response curves. The stability of the chemosensitivity was tested after 25-30 weeks continuous in vitro culture or prolonged storage in liquid nitrogen. One cell line underwent...... significant changes after continuous in vitro culture whereas the cell lines tested after prolonged storage in liquid nitrogen showed only minor changes. It is concluded that instead of considering the concentration necessary to achieve a certain degree of cell kill (e.g. ID50) in one experiment on one cell...

  19. Distribution of hydrogen within the HDR-containment under severe accident conditions. OECD standard problem. Final comparison report

    International Nuclear Information System (INIS)

    Karwat, H.

    1992-08-01

    The present report summarizes the results of the International Standard Problem Exercise ISP-29, based on the HDR Hydrogen Distribution Experiment E11.2. Post-test analyses are compared to experimentally measured parameters, well-known to the analysis. This report has been prepared by the Institute for Reactor Dynamics and Reactor Safety of the Technical University Munich under contract with the Gesellschaft fuer Anlagen- und Reaktorsicherheit (GRS) which received funding for this activity from the German Ministry for Research and Technology (BMFT) under the research contract RS 792. The HDR experiment E11.2 has been performed by the Kernforschungszentrum Karlsruhe (KfK) in the frame of the project 'Projekt HDR-Sicherheitsprogramm' sponsored by the BMFT. Ten institutions from eight countries participated in the post-test analysis exercise which was focussing on the long-lasting gas distribution processes expected inside a PWR containment under severe accident conditions. The gas release experiment was coupled to a long-lasting steam release into the containment typical for an unmitigated small break loss-of-coolant accident. In lieu of pure hydrogen a gas mixture consisting of 15% hydrogen and 85% helium has been applied in order to avoid reaching flammability during the experiment. Of central importance are common overlay plots comparing calculated transients with measurements of the global pressure, the local temperature-, steam- and gas concentration distributions throughout the entire HDR containment. The comparisons indicate relatively large margins between most calculations and the experiment. Having in mind that this exercise was specified as an 'open post-test' analysis of well-known measured data the reasons for discrepancies between measurements and simulations were extensively discussed during a final workshop. It was concluded that analytical shortcomings as well as some uncertainties of experimental boundary conditions may be responsible for deviations

  20. Analytical method for the identification and assay of 12 phthalates in cosmetic products: application of the ISO 12787 international standard "Cosmetics-Analytical methods-Validation criteria for analytical results using chromatographic techniques".

    Science.gov (United States)

    Gimeno, Pascal; Maggio, Annie-Françoise; Bousquet, Claudine; Quoirez, Audrey; Civade, Corinne; Bonnet, Pierre-Antoine

    2012-08-31

    Esters of phthalic acid, more commonly named phthalates, may be present in cosmetic products as ingredients or contaminants. Their presence as contaminant can be due to the manufacturing process, to raw materials used or to the migration of phthalates from packaging when plastic (polyvinyl chloride--PVC) is used. 8 phthalates (DBP, DEHP, BBP, DMEP, DnPP, DiPP, DPP, and DiBP), classified H360 or H361, are forbidden in cosmetics according to the European regulation on cosmetics 1223/2009. A GC/MS method was developed for the assay of 12 phthalates in cosmetics, including the 8 phthalates regulated. Analyses are carried out on a GC/MS system with electron impact ionization mode (EI). The separation of phthalates is obtained on a cross-linked 5%-phenyl/95%-dimethylpolysiloxane capillary column 30 m × 0.25 mm (i.d.) × 0.25 mm film thickness using a temperature gradient. Phthalate quantification is performed by external calibration using an internal standard. Validation elements obtained on standard solutions, highlight a satisfactory system conformity (resolution>1.5), a common quantification limit at 0.25 ng injected, an acceptable linearity between 0.5 μg mL⁻¹ and 5.0 μg mL⁻¹ as well as a precision and an accuracy in agreement with in-house specifications. Cosmetic samples ready for analytical injection are analyzed after a dilution in ethanol whereas more complex cosmetic matrices, like milks and creams, are assayed after a liquid/liquid extraction using ter-butyl methyl ether (TBME). Depending on the type of cosmetics analyzed, the common limits of quantification for the 12 phthalates were set at 0.5 or 2.5 μg g⁻¹. All samples were assayed using the analytical approach described in the ISO 12787 international standard "Cosmetics-Analytical methods-Validation criteria for analytical results using chromatographic techniques". This analytical protocol is particularly adapted when it is not possible to make reconstituted sample matrices. Copyright © 2012

  1. Mathematical models of cytotoxic effects in endpoint tumor cell line assays: critical assessment of the application of a single parametric value as a standard criterion to quantify the dose-response effects and new unexplored proposal formats.

    Science.gov (United States)

    Calhelha, Ricardo C; Martínez, Mireia A; Prieto, M A; Ferreira, Isabel C F R

    2017-10-23

    The development of convenient tools for describing and quantifying the effects of standard and novel therapeutic agents is essential for the research community, to perform more precise evaluations. Although mathematical models and quantification criteria have been exchanged in the last decade between different fields of study, there are relevant methodologies that lack proper mathematical descriptions and standard criteria to quantify their responses. Therefore, part of the relevant information that can be drawn from the experimental results obtained and the quantification of its statistical reliability are lost. Despite its relevance, there is not a standard form for the in vitro endpoint tumor cell lines' assays (TCLA) that enables the evaluation of the cytotoxic dose-response effects of anti-tumor drugs. The analysis of all the specific problems associated with the diverse nature of the available TCLA used is unfeasible. However, since most TCLA share the main objectives and similar operative requirements, we have chosen the sulforhodamine B (SRB) colorimetric assay for cytotoxicity screening of tumor cell lines as an experimental case study. In this work, the common biological and practical non-linear dose-response mathematical models are tested against experimental data and, following several statistical analyses, the model based on the Weibull distribution was confirmed as the convenient approximation to test the cytotoxic effectiveness of anti-tumor compounds. Then, the advantages and disadvantages of all the different parametric criteria derived from the model, which enable the quantification of the dose-response drug-effects, are extensively discussed. Therefore, model and standard criteria for easily performing the comparisons between different compounds are established. The advantages include a simple application, provision of parametric estimations that characterize the response as standard criteria, economization of experimental effort and enabling

  2. New approaches for the standardization and validation of a real-time qPCR assay using TaqMan probes for quantification of yellow fever virus on clinical samples with high quality parameters.

    Science.gov (United States)

    Fernandes-Monteiro, Alice G; Trindade, Gisela F; Yamamura, Anna M Y; Moreira, Otacilio C; de Paula, Vanessa S; Duarte, Ana Cláudia M; Britto, Constança; Lima, Sheila Maria B

    2015-01-01

    The development and production of viral vaccines, in general, involve several steps that need the monitoring of viral load throughout the entire process. Applying a 2-step quantitative reverse transcription real time PCR assay (RT-qPCR), viral load can be measured and monitored in a few hours. In this context, the development, standardization and validation of a RT-qPCR test to quickly and efficiently quantify yellow fever virus (YFV) in all stages of vaccine production are extremely important. To serve this purpose we used a plasmid construction containing the NS5 region from 17DD YFV to generate the standard curve and to evaluate parameters such as linearity, precision and specificity against other flavivirus. Furthermore, we defined the limits of detection as 25 copies/reaction, and quantification as 100 copies/reaction for the test. To ensure the quality of the method, reference controls were established in order to avoid false negative results. The qRT-PCR technique based on the use of TaqMan probes herein standardized proved to be effective for determining yellow fever viral load both in vivo and in vitro, thus becoming a very important tool to assure the quality control for vaccine production and evaluation of viremia after vaccination or YF disease.

  3. Nanoparticle-assay marker interaction: effects on nanotoxicity assessment

    International Nuclear Information System (INIS)

    Zhao, Xinxin; Xiong, Sijing; Huang, Liwen Charlotte; Ng, Kee Woei; Loo, Say Chye Joachim

    2015-01-01

    Protein-based cytotoxicity assays such as lactate dehydrogenase (LDH) and tumor necrosis factor-alpha (TNF-α) are commonly used in cytotoxic evaluation of nanoparticles (NPs) despite numerous reports on possible interactions with protein markers in these assays that can confound the results obtained. In this study, conventional cytotoxicity assays where assay markers may (LDH and TNF- α) or may not (PicoGreen and WST-8) come into contact with NPs were used to evaluate the cytotoxicity of NPs. The findings revealed selective interactions between negatively charged protein assay markers (LDH and TNF- α) and positively charged ZnO NPs under abiotic conditions. The adsorption and interaction with these protein assay markers were strongly influenced by surface charge, concentration, and specific surface area of the NPs, thereby resulting in less than accurate cytotoxic measurements, as observed from actual cell viability measurements. An improved protocol for LDH assay was, therefore, proposed and validated by eliminating any effects associated with protein–particle interactions. In view of this, additional measures and precautions should be taken when evaluating cytotoxicity of NPs with standard protein-based assays, particularly when they are of opposite charges

  4. Nanoparticle-assay marker interaction: effects on nanotoxicity assessment

    Science.gov (United States)

    Zhao, Xinxin; Xiong, Sijing; Huang, Liwen Charlotte; Ng, Kee Woei; Loo, Say Chye Joachim

    2015-01-01

    Protein-based cytotoxicity assays such as lactate dehydrogenase (LDH) and tumor necrosis factor-alpha (TNF-α) are commonly used in cytotoxic evaluation of nanoparticles (NPs) despite numerous reports on possible interactions with protein markers in these assays that can confound the results obtained. In this study, conventional cytotoxicity assays where assay markers may (LDH and TNF- α) or may not (PicoGreen and WST-8) come into contact with NPs were used to evaluate the cytotoxicity of NPs. The findings revealed selective interactions between negatively charged protein assay markers (LDH and TNF- α) and positively charged ZnO NPs under abiotic conditions. The adsorption and interaction with these protein assay markers were strongly influenced by surface charge, concentration, and specific surface area of the NPs, thereby resulting in less than accurate cytotoxic measurements, as observed from actual cell viability measurements. An improved protocol for LDH assay was, therefore, proposed and validated by eliminating any effects associated with protein-particle interactions. In view of this, additional measures and precautions should be taken when evaluating cytotoxicity of NPs with standard protein-based assays, particularly when they are of opposite charges.

  5. Nanoparticle-assay marker interaction: effects on nanotoxicity assessment

    Energy Technology Data Exchange (ETDEWEB)

    Zhao, Xinxin; Xiong, Sijing; Huang, Liwen Charlotte; Ng, Kee Woei, E-mail: kwng@ntu.edu.sg; Loo, Say Chye Joachim, E-mail: joachimloo@ntu.edu.sg [Nanyang Technological University, School of Materials Science and Engineering (Singapore)

    2015-01-15

    Protein-based cytotoxicity assays such as lactate dehydrogenase (LDH) and tumor necrosis factor-alpha (TNF-α) are commonly used in cytotoxic evaluation of nanoparticles (NPs) despite numerous reports on possible interactions with protein markers in these assays that can confound the results obtained. In this study, conventional cytotoxicity assays where assay markers may (LDH and TNF- α) or may not (PicoGreen and WST-8) come into contact with NPs were used to evaluate the cytotoxicity of NPs. The findings revealed selective interactions between negatively charged protein assay markers (LDH and TNF- α) and positively charged ZnO NPs under abiotic conditions. The adsorption and interaction with these protein assay markers were strongly influenced by surface charge, concentration, and specific surface area of the NPs, thereby resulting in less than accurate cytotoxic measurements, as observed from actual cell viability measurements. An improved protocol for LDH assay was, therefore, proposed and validated by eliminating any effects associated with protein–particle interactions. In view of this, additional measures and precautions should be taken when evaluating cytotoxicity of NPs with standard protein-based assays, particularly when they are of opposite charges.

  6. 20 CFR 667.274 - What health and safety standards apply to the working conditions of participants in activities...

    Science.gov (United States)

    2010-04-01

    ... WIA? (a) Health and safety standards established under Federal and State law otherwise applicable to... 20 Employees' Benefits 3 2010-04-01 2010-04-01 false What health and safety standards apply to the... I OF THE WORKFORCE INVESTMENT ACT Administrative Rules, Costs and Limitations § 667.274 What health...

  7. Condition Assessment Survey (CAS) Program. Deficiency standards and inspections methods manual: Volume 8, 0.08 Mechanical, Book 1

    Energy Technology Data Exchange (ETDEWEB)

    1993-05-01

    System information is presented for asset determinant factor/CAS repair codes/CAS cost factors; guide sheet too & material listing; testing methods; inspection frequency; standard system design life tables; and system work breakdown structure. Deficiency standards are given for plumbing, fire protection, heating, cooling, and special (drinking water cooling systems).

  8. The Impact of Statistical Adjustment on Conditional Standard Errors of Measurement in the Assessment of Physician Communication Skills

    Science.gov (United States)

    Raymond, Mark R.; Clauser, Brian E.; Furman, Gail E.

    2010-01-01

    The use of standardized patients to assess communication skills is now an essential part of assessing a physician's readiness for practice. To improve the reliability of communication scores, it has become increasingly common in recent years to use statistical models to adjust ratings provided by standardized patients. This study employed ordinary…

  9. Condition Assessment Survey (CAS) Program. Deficiency standards and inspections methods manual: Volume 7, 0.07 Conveying

    Energy Technology Data Exchange (ETDEWEB)

    1993-05-01

    System information is given for asset determinant factor/CAS repair codes/CAS cost factors; guide sheet tool & material listing; testing methods; inspection frequency; standard system design life tables; and system work breakdown structure. Deficiency standards and inspection methods are presented for elevators and special conveyors.

  10. Condition Assessment Survey (CAS) Program. Deficiency standards and inspections methods manual: Volume 3, 0.03 Superstructure

    Energy Technology Data Exchange (ETDEWEB)

    1993-05-01

    General information is presented on asset determinant factor/CAS profile codes/CAS cost process; guide sheet tool & material listing; testing methods; inspection frequency; standard system design life tables; system work breakdown structure; and general system/material data. Deficiency standards and inspection methods are presented for beams; pre-engineered building systems; floors; roof structure; stairs; and fireproofing.

  11. Electromagnetic Scattering Analysis of Coated Conductors With Edges Using the Method of Auxiliary Sources (MAS) in Conjunction With the Standard Impedance Boundary Condition (SIBC)

    DEFF Research Database (Denmark)

    Anastassiu, H.T.; D.I.Kaklamani, H.T.; Economou, D.P.

    2002-01-01

    A novel combination of the method of auxiliary sources (MAS) and the standard impedance boundary condition (SIBC) is employed in the analysis of transverse magnetic (TM) plane wave scattering from infinite, coated, perfectly conducting cylinders with square cross sections. The scatterer is initia......A novel combination of the method of auxiliary sources (MAS) and the standard impedance boundary condition (SIBC) is employed in the analysis of transverse magnetic (TM) plane wave scattering from infinite, coated, perfectly conducting cylinders with square cross sections. The scatterer...... efficient than the MoM/SIBC method, proving that the proposed novel combination is a powerful and advantageous computational tool....

  12. Analysis of blood coagulation in mice: pre-analytical conditions and evaluation of a home-made assay for thrombin-antithrombin complexes

    Directory of Open Access Journals (Sweden)

    Meijers Joost CM

    2005-08-01

    Full Text Available Abstract Background The use of mouse models for the study of thrombotic disorders has gained increasing importance. Methods for measurement of coagulation activation in mice are, however, scarce. The primary aim of this study was to develop a specific mouse thrombin-antithrombin (TAT ELISA for measurement of coagulation activation and to compare it with two commercially available assays for human TAT complexes. In addition, we aimed to improve methods for mouse plasma anticoagulation and preparation. Methods and results First, for the measurement of TAT-complexes in plasma a mouse specific TAT-ELISA was developed using rabbit polyclonal antibodies raised against mouse thrombin and rat antithrombin, respectively. This ELISA detected an increase in TAT levels in a mouse model of endotoxemia. Two commercial human TAT ELISAs appeared to be less specific for mouse thrombin-rat antithrombin complexes. Second, to prevent clotting of mouse blood sodium citrate was either mixed with blood during collection in a syringe or was injected intravenously immediately prior to blood collection. Intravenous sodium citrate completely inhibited blood coagulation resulting in plasma with consistently low TAT levels. Sodium citrate mixed with blood during collection resulted in increased TAT levels in 4 out of 16 plasma samples. Third, heparinase was added to plasma samples after in vivo injection of different heparin doses to test its neutralizing effect. Heparinase neutralized up to a 20 U of heparin/mouse and resulted in accurate APTT and factor VIII determinations. Conclusion These procedures and reagents for plasma preparation and coagulation testing will improve studies on thrombotic disorders in mice.

  13. Condition Assessment Survey (CAS) Program. Deficiency standards and inspections methods manual: Volume 1, 0.01 Foundations and footings

    Energy Technology Data Exchange (ETDEWEB)

    1993-05-01

    General information is presented for asset determinant factor/CAS repair codes/CAS cost factors; guide sheet tool & material listing; testing methods; inspection frequency; standard system design life tables; system work breakdown structure; and general system/material data. Deficiency standards and inspection methods are given for footings - spread/strip/grade beams; foundation walls; foundation dampproofing/waterproofing; excavation/backfill/ and piles & caissons.

  14. Condition Assessment Survey (CAS) Program. Deficiency standards and inspections methods manual: Volume 5, 0.05 Roofing

    Energy Technology Data Exchange (ETDEWEB)

    1993-05-01

    General information is presented for asset determinant factor/CAS repair codes/CAS cost factors; guide sheet tool & material listing; testing methods; inspection frequency; standard system design life tables; and system work breakdown structure. Deficiency standards and inspection methods are presented for built-up membrane; single- ply membrane; metal roofing systems; coated foam membrane; shingles; tiles; parapets; roof drainage system; roof specialties; and skylights.

  15. Condition Assessment Survey (CAS) Program. Deficiency standards and inspections methods manual: Volume 11, 0.11 Specialty systems

    Energy Technology Data Exchange (ETDEWEB)

    1993-05-01

    General information is presented for asset determinant factor/CAS repair codes/CAS cost factors; guide sheet tool & material listing; testing methods; inspection frequency; standard system design life tables; system work breakdown structure; and general system/material data. Deficiency standards and inspection methods are presented for canopies; loading dock systems; tanks; domes (bulk storage, metal framing); louvers & vents; access floors; integrated ceilings; and mezzanine structures.

  16. Condition Assessment Survey (CAS) Program. Deficiency standards and inspections methods manual: Volume 6, 0.06 Interior construction

    Energy Technology Data Exchange (ETDEWEB)

    1993-05-01

    General information is presented for asset determinant factor/CAS repair codes/CAS cost factors; guide sheet tool & material listing; testing methods; inspection frequency; standard system design life tables; system work breakdown structure; and general system/material data. Deficiency standards and inspection methods are presented for conventional and specialty partitions, toilet partitions & accessories, interior doors, paint finishes/coatings/ wall covering systems; floor finishing systems; and ceiling systems.

  17. Condition Assessment Survey (CAS) Program. Deficiency standards and inspections methods manual: Volume 12, 0.12 Sitework

    Energy Technology Data Exchange (ETDEWEB)

    1993-05-01

    General information is presented for asset determinant factor/CAS repair codes/CAS cost factors; guide sheet tool & material listing; testing methods; inspection frequency; standard system design life tables; system work breakdown structure; and general system/material data. Deficiency standards and inspection methods are given for utility distribution systems, central heating, central cooling, electrical, utility support structures, paving roadways/walkways, and tunnels.

  18. The effect of milk components and storage conditions on the virulence of Listeria monocytogenes as determined by a Caco-2 cell assay.

    Science.gov (United States)

    Pricope-Ciolacu, Luminita; Nicolau, Anca Ioana; Wagner, Martin; Rychli, Kathrin

    2013-08-16

    Nearly all cases of human listeriosis have been associated with consumption of contaminated food, therefore the investigation of the virulence of Listeria (L.) monocytogenes after exposure to environmental conditions in food matrices is critical in order to understand and control its impact on public health. As milk and dairy products have been implicated in more than half of the listeriosis outbreaks, we investigated the in vitro virulence of L. monocytogenes incubated in different milk types at various storage conditions. Incubation in pasteurized milk at refrigeration conditions (4°C) revealed a higher invasion and intracellular proliferation of four different L. monocytogenes strains compared to raw milk using human intestinal epithelial Caco-2 cells. Furthermore the period of storage, which increased L. monocytogenes cell numbers, decreased in vitro virulence. However, L. monocytogenes stored for 3weeks at 4°C in milk are still able to invade and proliferate into the host cell. Interestingly abused storage temperatures (25°C and 30°C) for a short time period (2h) revealed an attenuated impact on the in vitro virulence of L. monocytogenes compared to the storage temperature of 4°C. Regarding the major milk compounds, the level of milk fat significantly affected the in vitro virulence of L. monocytogenes. Pre-incubation in milk with high fat content (3.6%) resulted in a lower invasion capability compared to milk with low fat content. In contrast casein and lactose did not influence the invasiveness of L. monocytogenes into the host cell. In conclusion our study shows that the milk environment and different storage conditions influence the in vitro virulence of L. monocytogenes, both of which have to be considered in the risk assessment of contaminated food. Copyright © 2013 Elsevier B.V. All rights reserved.

  19. Development and evaluation of a loop-mediated isothermal amplification assay for the rapid detection of porcine cytomegalovirus under field conditions

    Directory of Open Access Journals (Sweden)

    Yang Jin-Long

    2012-12-01

    Full Text Available Abstract Background Porcine cytomegalovirus (PCMV induces silent infection in adult pigs but more frequently causes fatal, generalized infection in newborn piglets. This study aimed to develop a new loop-mediated isothermal amplification (LAMP method for the sensitive, rapid, and inexpensive detection of PCMV under field conditions. Methods Tissue obtained from nine-week-old PCMV-free Landrace pigs or pig samples from postmortem examinations were analyzed. The samples were found to have clinical signs and lesions consistent with inclusion body rhinitis. Six specific primers were designed by targeting the PCMV DNA polymerase (DPOL DNA. The LAMP reaction was optimized in a water bath. The sensitivity and specificity of LAMP and polymerase chain reaction (PCR were compared. Results PCMV DNA was amplified at 65°C, and the result could be detected as early as 30 min into the reaction. Positive reactions could be visualized by the naked eye as a color change brought on by the addition of SYBR Green. The sensitivity and specificity of LAMP were found to be similar to those of the PCR. Conclusions LAMP is a high-throughput technique for the detection of PCMV and has a high specificity, sensitivity and simplicity; these factors make it suitable for detection of PCMV under field conditions.

  20. Evaluation of environmental and economic effectiveness of the Cross Compliance 4.3 Standards "Maintenance of olive groves and vineyards in good vegetative conditions"

    Directory of Open Access Journals (Sweden)

    Luigi Sansone

    2015-11-01

    Full Text Available This paper reports the first observations made in three farms of the Council for Agricultural Research and Economics (CREA relating to the environmental monitoring of the standard 4.3 maintenance of olive groves and vineyards in good vegetative conditions and analysis of differential of competitiveness  for both crops.

  1. ASME N511-19XX, Standard for periodic in-service testing of nuclear air treatment, heating, ventilating and air conditioning systems

    International Nuclear Information System (INIS)

    1997-01-01

    A draft version of the Standard is presented in this document. The Standard covers the requirements for periodic in-service testing of nuclear safety-related air treatment, heating, ventilating, and air conditioning systems in nuclear facilities. The Standard provides a basis for the development of test programs and does not include acceptance criteria, except in cases where the results of one test influence the performance of other tests. The Standard covers general inspection and test requirements, reference values, inspection and test requirements, generic tests, acceptance criteria, in-service test requirements, testing following an abnormal incident, corrective action requirements, and quality assurance. Mandatory appendices provide a visual inspection checklist and four test procedures. Non-mandatory appendices provide additional information and guidance on mounting frame pressure leak test procedure, corrective action, challenge gas substitute selection criteria, and test program development. 8 refs., 10 tabs

  2. 40 CFR 80.553 - Under what conditions may the small refiner gasoline sulfur standards be extended for a small...

    Science.gov (United States)

    2010-07-01

    ... gasoline produced by the refinery must meet the gasoline sulfur standards under subpart H of this Part as... all succeeding compliance periods and all gasoline produced by the refinery must meet the gasoline... applicable). Upon such effective date, all gasoline produced by the refiner must meet the gasoline sulfur...

  3. 13 CFR 107.700 - Compliance with size standards in part 121 of this chapter as a condition of Assistance.

    Science.gov (United States)

    2010-01-01

    ... Assistance SMALL BUSINESS ADMINISTRATION SMALL BUSINESS INVESTMENT COMPANIES Financing of Small Businesses by... assistance and management services only to a Small Business. To determine whether an applicant is a Small... 13 Business Credit and Assistance 1 2010-01-01 2010-01-01 false Compliance with size standards in...

  4. Estimation of Sensitivity and Specificity of Three Conditionally Dependent Diagnostic Tests in the Absence of a Gold Standard

    NARCIS (Netherlands)

    Engel, B.; Swildens, B.; Stegeman, J.A.; Buist, W.G.; Jong, de M.

    2006-01-01

    This article presents a model to evaluate the accuracy of diagnostic tests. Data from three tests for the detection of EF-positive Streptococcus suis serotype 2 strains in sows were analyzed. The data were collected in a field study in the absence of a gold standard, that is, the true disease status

  5. Individual condition, standard metabolic rate, and rearing temperature influence steelhead and rainbow trout (Oncorhynchus mykiss) life histories

    Science.gov (United States)

    Matthew R. Sloat; Gordon H. Reeves

    2014-01-01

    We reared juvenile Oncorhychus mykiss with low and high standard metabolic rates (SMR) under alternative thermal regimes to determine how these proximate factors influence life histories in a partially migratory salmonid fish. High SMR significantly decreased rates of freshwater maturation and increased rates of smoltification in females, but not...

  6. Condition Assessment Survey (CAS) Program. Deficiency standards and inspections methods manual: Volume 2, 0.02 Substructure

    Energy Technology Data Exchange (ETDEWEB)

    1993-05-01

    System information is given for asset determinant factor/CAS repair codes/CAS cost factors; guide sheet tool & material listing; testing methods; inspection frequency; standard system design life tables; system work breakdown structure; and general system/material data. System assembly/component deficiencies and inspection methods are given for slabs-on-grade, columns, and column fireproofing.

  7. Different Assay Conditions for Detecting the Production and Release of Heat-Labile and Heat-Stable Toxins in Enterotoxigenic Escherichia coli Isolates

    Directory of Open Access Journals (Sweden)

    Letícia B. Rocha

    2013-12-01

    Full Text Available Enterotoxigenic Escherichia coli (ETEC produce heat-labile (LT and/or heat-stable enterotoxins (ST. Despite that, the mechanism of action of both toxins are well known, there is great controversy in the literature concerning the in vitro production and release of LT and, for ST, no major concerns have been discussed. Furthermore, the majority of published papers describe the use of only one or a few ETEC isolates to define the production and release of these toxins, which hinders the detection of ETEC by phenotypic approaches. Thus, the present study was undertaken to obtain a better understanding of ST and LT toxin production and release under laboratory conditions. Accordingly, a collection of 90 LT-, ST-, and ST/LT-producing ETEC isolates was used to determine a protocol for toxin production and release aimed at ETEC detection. For this, we used previously raised anti-LT antibodies and the anti-ST monoclonal and polyclonal antibodies described herein. The presence of bile salts and the use of certain antibiotics improved ETEC toxin production/release. Triton X-100, as chemical treatment, proved to be an alternative method for toxin release. Consequently, a common protocol that can increase the production and release of LT and ST toxins could facilitate and enhance the sensitivity of diagnostic tests for ETEC using the raised and described antibodies in the present work.

  8. Aid conditionalities, international Good Manufacturing Practice standards and local production rights: a case study of local production in Nepal.

    Science.gov (United States)

    Brhlikova, Petra; Harper, Ian; Subedi, Madhusudan; Bhattarai, Samita; Rawal, Nabin; Pollock, Allyson M

    2015-06-14

    Local pharmaceutical production has been endorsed by the WHO as a means of addressing health priorities of developing countries. However, local producers of essential medicines must comply with international pharmaceutical standards in order to be eligible to compete in donor tenders. These standards determine production rights for on-patent and off-patent medicines, and guide international procurement of medicines. We reviewed the literature on the impact of Good Manufacturing Practice (GMP) on local production; a gap analysis from the literature review indicated a need for further research. Over sixty interviews were conducted with people involved in the Nepali pharmaceutical production and distribution chain from 2006 to 2009 on the GMP areas of relevance: regulatory capacity, staffing, funding and training, resourcing of GMP, inspectors' interpretation of the rules and compliance. Although Nepal producers have increased their overall share of the domestic market, only the public manufacturer, Royal Drugs, focuses on medicines for public health programmes; private producers engage mainly in brand competition for private markets, not essential medicines. Nepali regulators and producers state that implementation of GMP standards is hindered by low regulatory capacity, insufficient training of staff in the industry, financial constraints and lack of investment for upgrading capital. The transition period to mandatory compliance with WHO GMP rules is lengthy. Less than half of private producers had WHO GMP in 2013. Producers are not directly affected by international harmonisation of standards as they do not export medicines and the Nepali regulator does not enforce the WHO standards strictly. Without an international GMP certificate they cannot tender for donor dependent health programmes. In Nepal, local private manufacturers focus mainly on brand competition for private consumption not essential medicines, the government preferentially procures essential

  9. Influence of biological assay conditions on stability assessment of radiometal-labelled peptides exemplified using a {sup 177}Lu-DOTA-minigastrin derivative

    Energy Technology Data Exchange (ETDEWEB)

    Ocak, Meltem [Clinical Department of Nuclear Medicine, Medical University Innsbruck, A-6020, Innsbruck (Austria); Department of Pharmaceutical Technology, Pharmacy Faculty, Istanbul University, 34116, Istanbul (Turkey); Helbok, Anna; Guggenberg, Elisabeth von [Clinical Department of Nuclear Medicine, Medical University Innsbruck, A-6020, Innsbruck (Austria); Ozsoy, Y. [Department of Pharmaceutical Technology, Pharmacy Faculty, Istanbul University, 34116, Istanbul (Turkey); Kabasakal, Levent [Department of Nuclear Medicine, Cerrahpasa Medical Faculty, 34098, Istanbul (Turkey); Kremser, Leopold [Division of Clinical Biochemistry, Protein Micro-Analysis Facility, Biocenter, Medical University Innsbruck, A-6020, Innsbruck (Austria); Decristoforo, Clemens, E-mail: clemens.decristoforo@uki.a [Clinical Department of Nuclear Medicine, Medical University Innsbruck, A-6020, Innsbruck (Austria)

    2011-02-15

    Introduction: Lack of correlation between in vitro and in vivo stability is a general problem for the development of radiopeptides especially in the case of minigastrin derivatives for therapeutic applications. In this study, we compared the influence of experimental conditions on radiopeptide stability results in vitro using a model Minigastrin (MG) analogue labelled with Lu-177. Additionally, we attempted to characterize the main serum enzymatic cleavage sites by matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) analysis. Methods: In vitro stability of a DOTA-minigastrin derivative ({sup 177}Lu-DOTA-His-His-Glu-Ala-Tyr-Gly-Trp-NIe-Asp-Phe-NH{sub 2}) was tested in serum, rat tissue homogenates and two different standardised enzymatic mixtures. Quantification of the metabolised radiopeptides at different time intervals was performed using reversed-phase high-performance liquid chromatography (RP-HPLC). Metabolites were characterised by MALDI-TOF-MS. Urine was collected after 15 min p.i. into the mice and compared with in vitro metabolites by RP-HPLC. Results: Faster degradation of the radiopeptide was found in blood in comparison with plasma and serum incubation and in components from rats faster than from human origin. Fast degradation was observed in kidney and liver homogenates as well as in standardised enzymatic mixtures, also revealing variations in the metabolic profile. In urine, no intact peptide was detected already 5 min post injection. MALDI-TOF-MS revealed major cleavage sites at the carboxy terminus of the peptide. Conclusion: Very variable results may be found when different kind of incubation media for testing radiopeptide stabilities is used. Serum incubation studies may overestimate stability; therefore, results should be interpreted with care and combined with alternative in vitro and in vivo investigations.

  10. Assay for applying super absorbent polymer in a low input corn (Zea mays L. production system aimed to reduce drought stress under Mashhad conditions

    Directory of Open Access Journals (Sweden)

    M. Jahan

    2016-05-01

    Full Text Available In order to investigate the effects of super absorbent polymer application on reduction of drought stress to corn, a split plot arrangement based on randomized complete block design with three replications was conducted at Research Field of Agriculture Faculty of Ferdowsi University of Mashhad during growing season of 2010-11. The main plot treatments were 1 application of 40 kg.ha-1 super absorbent, 2 application of 80 kg.ha-1 super absorbent and 3 no application of super absorbent polymer. Three irrigation intervals (7, 10 and 14 days assigned to sub plots. The results showed that super absorbent application affected plant height (H, and dry matter production (DM as the highest of these traits resulted from level 2 of super absorbent application (140.5 cm, and 144.5 g.m-2, respectively. H, DM, canopy temperature (CT, cob number (N, fresh yield (FY, economic yield (EY and 100-seed weight affected by irrigation intervals. There was no significant difference between 10 and 14 days irrigation interval as H, DM, CT, harvest Index (HI and 100-seed weight, these results could be important concerning to reduce used water to irrigate corn. As experimental treatments did not have any effect on Leaf Area Index (LAI, and HI, it seems the positive effects of treatments revealed due to improved soil water holding capacity, soil physical properties improvement and reduction of drought stress. Interaction between super absorbent and irrigation intervals indicates that by level 2 super absorbent applications there are no significant differences between 14 and 10 days irrigation intervals, considering all traits. The same interaction just as before happened for 7 and 14 days irrigation intervals, except of EY and DM. In the other hand, by increasing application of super absorbent it could be possible to increase corn irrigation intervals from 7 to 14 days in Mashhad conditions without any reduction in yield and yield components. In general, these results

  11. Optimization of invertase assay conditions in rubber tree plants (Hevea brasiliensis Muell. Arg. Otimização das condiçõess do ensaio da invertase em seringueira (Hevea brasiliensis Muell. Arg.

    Directory of Open Access Journals (Sweden)

    Daria Pimenta de Oliveira

    2006-10-01

    Full Text Available The objective of this work was to define the optimal conditions for invertase assay, seeking to determine the ideal parameters for the different isoenzymes of leaf and bark tissues in adult rubber trees. Assays of varying pH, sucrose concentration and temperature of the reaction medium were conducted for the two investigated isoenzymes. The results pointed out the existence of two different pH related isoforms for the two analyzed tissues, with an isoenzyme being more active at pH 5,5 and the other at neutral/alkaline pH. Leaf blade isoenzymes presented similar values for substrate concentration, whereas the bark isoenzyme presented maximum values below those previously reported. The assays at different temperatures presented similar values for leaf isoenzymes, though they have differed significantly among the obtained values.O objetivo deste trabalho foi definir as condições ótimas para a realização do ensaio enzimático da invertase, procurando-se determinar os parâmetros ideais para as diferentes isoenzimas de tecidos foliares e da casca de plantas adultas de seringueira. Foram realizados ensaios variando-se o pH, a concentração da sacarose e a temperatura do meio de reação para as duas isoenzimas estudadas. Os resultados indicaram a existência de duas isoformas diferentes em relação ao pH nos dois tecidos analisados, sendo uma isoenzima mais ativa a pH 5,5 e outra em pH neutro/alcalino. Com relação à concentração do substrato, as isoenzimas da lâmina foliar apresentaram valores semelhantes, enquanto a isoenzima da casca, valores máximos inferiores aos observados anteriormente. Os ensaios conduzidos em diferentes temperaturas tiveram valores semelhantes nas isoenzimas da folha, embora tenham diferido significativamente entre dos valores obtidos.

  12. Salmonella detection in poultry meat and meat products by the Vitek immunodiagnostic assay system easy Salmonella method, a LightCycler polymerase chain reaction system, and the International Organization for Standardization method 6579.

    Science.gov (United States)

    Temelli, S; Eyigor, A; Carli, K T

    2012-03-01

    This study was conducted to evaluate the capability of the Vitek immunodiagnostic assay system easy Salmonella (VIDAS ESLM) method and a specific real-time PCR system (LightCycler, LCPCR) to complement the International Organization for Standardization Method 6579 (ISO) in detecting Salmonella from a total of 105 naturally contaminated samples comprised of poultry meat and poultry meat products. The detection limit of ISO and LCPCR was 9 cfu/mL for both poultry meat and poultry meat products, whereas that of VIDAS ESLM with both sample types was determined to be 90 cfu/mL. Twelve (33.33%), 11 (30.55%), and 18 (50.00%) out of 36 poultry meat samples were positive for Salmonella by ISO, VIDAS ESLM, and LCPCR, respectively. Salmonella detection rates from poultry meat products were 5.80% for ISO and 8.69% for LCPCR, whereas none of these products tested positive by VIDAS ESLM. In poultry meat samples, VIDAS ESLM and LCPCR detection results were in substantial agreement with ISO, with the relative accuracy, sensitivity, and specificity rates of 97.2, 91.7, and 100%, respectively, for VIDAS ESLM and 83.3, 100, and 75%, respectively, for LCPCR. This is the first report on the evaluation of both VIDAS ESLM and LCPCR to complement ISO for the rapid detection of Salmonella in poultry meat and meat products. We determined that both VIDAS ESLM and LCPCR have the potential to complement the ISO standard culture method in the rapid screening of Salmonella from naturally contaminated poultry meats. For the poultry meat products, VIDAS ESLM and LCPCR can be used for rapid primary screening, and they should be complemented absolutely by ISO. Although LCPCR can preferentially be used for initial screening poultry meat products, the results should definitely be confirmed by ISO. Also, the VIDAS ESLM did not seem to be a suitable method for detecting Salmonella in poultry meat products.

  13. Equipment qualification NPP in harsh environmental conditions occurring during a design basis accident (DBA), beyond design basis accident (BDBA) in accordance with international standards

    International Nuclear Information System (INIS)

    Aparkin, F.M.; )

    2015-01-01

    The purpose of Equipment Qualification NPP (EQ), safety-related electrical and mechanical equipment is: reducing the probability of equipment failure common cause in connection with the harsh environmental conditions at the design and beyond design basis accidents; to demonstrate that the safety related electrical and mechanical equipment can perform their specific functions related safety in harsh environments in design and beyond design basis accidents. Presentation defines methodological bases of practical EQ in accordance with international standards used for PWR, for demonstration the performance of its safety functions, which subjected to abnormal and accident conditions, including loss of ventilation systems, breaks feed waterline, steam and cooling water main system and seismic events [ru

  14. A fluorimetric assay for cortisol.

    Science.gov (United States)

    Appel, Daniel; Schmid, Rolf D; Dragan, Calin-Aurel; Bureik, Matthias; Urlacher, Vlada B

    2005-09-01

    A simple, rapid and sensitive fluorimetric assay for the quantitative determination of cortisol is reported. The assay is based on the formation of a fluorescent dye when cortisol is incubated with a mixture of sulfuric acid and acetic acid. The fluorescence spectrum recorded for the resulting dye shows a maximum extinction at 475 nm and a maximum emission at 525 nm. The solvent 2-methyl-4-pentanone was used for extraction and was found to act as a fluorescence amplifier. A limit of detection of 2.7 muM was achieved, making it possible to forego solvent evaporation. The assay suffers minor interference from 11-deoxycortisol which exhibits low fluorescence at lambda (ex): 460 nm; lambda (em): 505 nm. Typical standard deviations were below 4%. We validated the assay using a biotransformation with recombinant Schizosaccharomyces pombe which regioselectively hydroxylates 11-deoxycortisol to cortisol. The method described herein is suitable for preliminary screening of microorganisms capable of steroid hydroxylation.

  15. 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.

  16. Comparison of EHR-based diagnosis documentation locations to a gold standard for risk stratification in patients with multiple chronic conditions.

    Science.gov (United States)

    Martin, Shelby; Wagner, Jesse; Lupulescu-Mann, Nicoleta; Ramsey, Katrina; Cohen, Aaron; Graven, Peter; Weiskopf, Nicole G; Dorr, David A

    2017-08-02

    To measure variation among four different Electronic Health Record (EHR) system documentation locations versus 'gold standard' manual chart review for risk stratification in patients with multiple chronic illnesses. Adults seen in primary care with EHR evidence of at least one of 13 conditions were included. EHRs were manually reviewed to determine presence of active diagnoses, and risk scores were calculated using three different methodologies and five EHR documentation locations. Claims data were used to assess cost and utilization for the following year. Descriptive and diagnostic statistics were calculated for each EHR location. Criterion validity testing compared the gold standard verified diagnoses versus other EHR locations and risk scores in predicting future cost and utilization. Nine hundred patients had 2,179 probable diagnoses. About 70% of the diagnoses from the EHR were verified by gold standard. For a subset of patients having baseline and prediction year data (n=750), modeling showed that the gold standard was the best predictor of outcomes on average for a subset of patients that had these data. However, combining all data sources together had nearly equivalent performance for prediction as the gold standard. EHR data locations were inaccurate 30% of the time, leading to improvement in overall modeling from a gold standard from chart review for individual diagnoses. However, the impact on identification of the highest risk patients was minor, and combining data from different EHR locations was equivalent to gold standard performance. The reviewer's ability to identify a diagnosis as correct was influenced by a variety of factors, including completeness, temporality, and perceived accuracy of chart data.

  17. Determination of Ergot Alkaloids: Purity and Stability Assessment of Standards and Optimization of Extraction Conditions for Cereal Samples

    DEFF Research Database (Denmark)

    Krska, R.; Berthiller, F.; Schuhmacher, R.

    2008-01-01

    considerably above 98% apart from ergocristinine (94%), ergosine (96%), and ergosinine (95%). Also discussed is the optimization of extraction conditions presented in a recently published method for the quantitation of ergot alkaloids in food samples using solid-phase extraction with primary secondary amine...... (PSA) before LC/MS/MS. Based on the results obtained from these optimization studies, a mixture of acetonitrile with ammonium carbonate buffer was used as extraction solvent, as recoveries for all analyzed ergot alkaloids were significantly higher than those with the other solvents. Different sample......-solvent ratios and extraction times showed just minor influences in extraction efficacy. Finally, the stability of the ergot alkaloids in both raw cereals and cereal-based processed food extracts was studied. According to these studies, extracts should be prepared and analyzed the same day or stored below...

  18. Harmonization of radiobiological assays: why and how?

    International Nuclear Information System (INIS)

    Prasanna, Pataje G.

    2014-01-01

    The International Atomic Energy Agency has made available a technical manual for cytogenetic biodosimetry assays (dicentric chromosome aberration (DCA) and cytokinesis-block micronucleus (CBMN) assays) used for radiation dose assessment in radiation accidents. The International Standardization Organization, which develops standards and guidelines, also provides an avenue for laboratory accreditation, has developed guidelines and recommendations for performing cytogenetic biodosimetry assays. Harmonization of DCA and CBMN assays, has improved their accuracy. Double-blinded inter-laboratory comparison studies involving several networks have further validated DCA and CBMN assays and improved the confidence in their potential use for radiation dose assessment in mass casualties. This kind of international harmonization is lacking for pre-clinical radiobiology assays. The widely used pre-clinical assays that are relatively important to set stage for clinical trials include clonogenic assays, flow-cytometry assays, apoptotic assays, and tumor regression and growth delay assays. However, significant inter-laboratory variations occur with respect to data among laboratories. This raises concerns on the reliability and reproducibility of preclinical data that drives further development and translation. Lack of reproducibility may stem from a variety of factors such as poor scientist training, less than optimal experimental design, inadequate description of methodology, and impulse to publish only the positive data etc. Availability of technical manuals, standard operating procedures, accreditation avenues for laboratories performing such assays, inter-laboratory comparisons, and use of standardized protocols are necessary to enhance reliability and reproducibility. Thus, it is important that radiobiological assays are harmonized for laboratory protocols to ensure successful translation of pre-clinical research on radiation effect modulators to help design clinic trials with

  19. Determination of Nitrate Reductase Assay Depending on the Microbial Growth

    International Nuclear Information System (INIS)

    El-Kabbany, H.M.

    2012-01-01

    A rapid micro-dilution assay for determination of the antimicrobial susceptibility of different bacterial isolates was developed. This assay is based on the ability of the most of viable organisms to reduce nitrates. The MIC or MBC could be determined by nitrate reductase (NR) only after 30 to 90 min of incubation depending on the behaviour of microbial growth. Bacterial viability is detected by a positive nitrite reduction rather than visible turbidity. The nitrate reduction assay was compared with standard micro-assay using 250 isolates of different taxa against 10 antibiotics belonging to different classes. An excellent agreement of 82.5 % was found between the two methods and only 17.5 % of 1794 trials showed difference in the determined MIC by tow-dilution interval above or below the MIC determined by the turbidimetric method under the same test conditions. However, the nitrate reduction assay was more rapid and sensitive in detecting viable bacteria and so, established an accurate estimate of the minimal inhibitory concentration (MIC) or the minimal bacterial concentration (MBC). The nitrate reduction assay offers the additional advantage that it could be used to determine the MBC without having to subculture the broth. 232 cases of resistance were detected by NR and 4 different media were tested for susceptibility test. The bacterial isolates were exposed to ultra violet (UV) light for different period

  20. THE FORMS OF EDUCATIONAL COOPERATION IN SCHOOL UNDER THE CONDITIONS OF IMPLEMENTATION OF THE FEDERAL STATE EDUCATIONAL STANDARDS

    Directory of Open Access Journals (Sweden)

    L. G. Puzep

    2016-01-01

    Full Text Available The aim of the publication is to show possibilities of implementation of requirements of the federal state educational standard (FSES for formati on of the universal educational activities (UEA during group work at lessons at comprehensive school.Methods. Theoretical methods involve analysis, comparison and generalizationmethods; empirical method of interview.Results. The main components of learning technology in cooperation and stages of technological process of group work are presented. The features of the formation of small groups are described due to the criteria of duration of a group work and the age of pupils. The possible options for the role structure of discussion groups and ways of distribution of social roles in groups are found out. The authors show the main steps of the process of group work process and organization techniques of group work of students at different stages of the lesson that allow purposefully organize the situation of communication and reflective activities. Scientific novelty. The essence of an educational cooperation as forms of group interaction of the teacher and pupils at a lesson is considered in the context of FSES of the last generation. The emergent problems in the course of the group activities organization at school are disclosed. The authors propose their own opinion on the implementation of the basic stages of group work, which are defined as a technological lesson plan. In addition, the components of a technological lesson plan include the universal learning activities forming during groupactivities which are important for the implementation of FSES requirements.Practical significance. The proposed techniques for organizing group work of pupils at different stages of the lesson and a technological lesson plan describing the universal learning activities allow teachers to effectively use technology of cooperation in education, aimed at the formation of pupils’ communicative learning activities.

  1. Isotope tracing enhancement of chemiluminescence assays for nitric oxide research.

    Science.gov (United States)

    Cornelius, Julia; Tran, Tuan; Turner, Nicole; Piazza, Abigail; Mills, Lauren; Slack, Ryan; Hauser, Sean; Alexander, J Steven; Grisham, Matthew B; Feelisch, Martin; Rodriguez, Juan

    2009-02-01

    Chemiluminescence assays are used widely for the detection of nitric oxide (NO)-derived species in biological fluids and tissues. Here, we demonstrate that these assays can be interfaced with mass-sensitive detectors for parallel determination of isotopic abundance. Results obtained with tri-iodide and ascorbic acid-based reductive assays indicate that mass spectrometric detection enables NO isotope-tracing experiments to be carried out to a limit of detectability of a few picomoles, a sensitivity similar to that of standard gas phase chemiluminescence methods. The advantage afforded by mass spectrometric detection is demonstrated using the murine macrophage cell line J774, which is shown here to reduce 15NO3- to 15NO2- under anoxic conditions. The particular combination of an analytical and cellular system described here may hold promise for future characterization of the enzymatic pathways contributing to mammalian nitrate reductase activity, without background interference from 14NO2- derived from other sources.

  2. Standard test method for conducting friction tests of piston ring and cylinder liner materials under lubricated conditions

    CERN Document Server

    American Society for Testing and Materials. Philadelphia

    2011-01-01

    1.1 This test method covers procedures for conducting laboratory bench-scale friction tests of materials, coatings, and surface treatments intended for use in piston rings and cylinder liners in diesel or spark-ignition engines. The goal of this procedure is to provide a means for preliminary, cost-effective screening or evaluation of candidate ring and liner materials. A reciprocating sliding arrangement is used to simulate the contact that occurs between a piston ring and its mating liner near the top-dead-center position in the cylinder where liquid lubrication is least effective, and most wear is known to occur. Special attention is paid to specimen alignment, running-in, and lubricant condition. 1.2 This test method does not purport to simulate all aspects of a fired engine’s operating environment, but is intended to serve as a means for preliminary screening for assessing the frictional characteristics of candidate piston ring and liner material combinations in the presence of fluids that behave as u...

  3. Bifacial solar cell measurements under standard test conditions and the impact on cell-to-module loss analysis

    Science.gov (United States)

    Singh, Jai Prakash; Chai, Jing; Hsian Saw, Min; Khoo, Yong Sheng

    2017-08-01

    Bifacial cells are conventionally measured using gold-plated chuck, which is conductive and reflective. This measurement setup does not portray the actual operating conditions of the bifacial cells in a module. The reflective chuck causes an overestimation of the current due to the cell transmittance for the infrared light. The conductive chuck creates a shorter current flow path in the rear side of the cell and causes an over inflation of the fill factor measurement. In this study, we characterize and quantitatively analyze the difference between the bifacial cell measurements on different mounting chucks and calculate the cell-to-module (CTM) loss. To characterize the optical behavior of the bifacial cell and module, we perform external quantum efficiency, reflectance and transmittance measurements. The electrical behavior of the bifacial cell is studied using in-house developed software Griddler. Using Griddler, we calculate the difference in the fill factor of the bifacial cell due to the measurement using a conductive and non-conductive chuck, and estimate the corresponding CTM resistive losses.

  4. Contribution to the standardization of the chromatographic conditions for the lipophilicity assessment of neutral and basic drugs

    Energy Technology Data Exchange (ETDEWEB)

    Giaginis, Costas [Department of Pharmaceutical Chemistry, School of Pharmacy, University of Athens, Panepistimiopolis, Zografou, Athens 15771 (Greece); Department of Forensic Medicine and Toxicology, Medical School, University of Athens, 75 Mikras Asias Street, Athens 11527 (Greece); Theocharis, Stamatios [Department of Forensic Medicine and Toxicology, Medical School, University of Athens, 75 Mikras Asias Street, Athens 11527 (Greece); Tsantili-Kakoulidou, Anna [Department of Pharmaceutical Chemistry, School of Pharmacy, University of Athens, Panepistimiopolis, Zografou, Athens 15771 (Greece)]. E-mail: tsantili@pharm.uoa.gr

    2006-07-28

    The chromatographic conditions aiming to a better simulation of n-octanol-water partitioning using a base deactivated silica (BDS) column as stationary phase were investigated for structurally diverse basic and neutral drugs. Extrapolated retention factors log k{sub w}, determined using different methanol fractions as organic modifier, were considered as lipophilicity indices. The effect of n-decylamine and n-octanol as mobile phase additives was examined and the appropriateness of the final retention outcome to reproduce lipophilicity data was evaluated. Moreover, the influence of n-octanol on the linearity of the log k/methanol fraction relationship and on the uniformity of the retention mechanism was investigated. 1:1 correlation between log k{sub w} values and the logarithm of the distribution coefficient (log D) was established for basic drugs in presence of both n-decylamine and n-octanol as mobile phase additives. However, for neutral drugs n-decylamine proved to be a sufficient and more important factor than n-octanol.

  5. Effectiveness of the GAEC cross compliance standard Maintenance of olive groves in good vegetative condition in avoiding the deterioration of habitats and land abandonment

    Directory of Open Access Journals (Sweden)

    Elena Santilli

    2011-08-01

    Full Text Available The last CAP reform (Council Regulation (EC n. 1782/2003, coincided with the mandatory obligations of the principles of cross compliance, under which all compensatory payments given in the context of the former reform packages were replaced by a Single Payment Scheme (SPS, bound to fulfillment of certain requirements and minimum standards regarding the environment and animal welfare, as well as maintaining the land in good agricultural and environmental conditions. For the olive sector, where potential risks are mainly associated to the abandonment of groves in marginal areas with consequent negative environmental impact, it has been specifically established the standard 4.3 of the Good Agricultural and Environmental Conditions (GAEC which concerns the Maintenance of olive groves and vines in good vegetative conditions. This GAEC standard was formulated to ensure a minimum level of land maintenance and to avoid the deterioration of habitats. To achieve these objectives it should be considered that a good vegetative development is strictly related to the care of the soil in which the plants grow. Erosion, organic matter and soil structure decay are the most commonly identified impacts for olive orchards, 30% of which are localized in areas with difficult orographic conditions. In this sense, proper hydraulic and mechanical techniques, cover cropping, green manuring and timely pruning turns, are essential to minimize losses due to soil erosion, to limit the leaching of nutrients and to maintain the plant productivity. Furthermore, grinded pruning residues should be spread in situ and weeds, watersprouts and suckers should be periodically cut off in order to increase the atmospheric CO2 sequestration and soil organic matter (OM and to prevent wildfires risk and nutrients competition. The application of the standard 4.3 requires further investigations, because, while numerous studies have shown that pruning is essential for the production, there are

  6. Standard partial molar heat capacities and enthalpies of formation of aqueous aluminate under hydrothermal conditions from integral heat of solution measurements

    International Nuclear Information System (INIS)

    Coulier, Yohann; Tremaine, Peter R.

    2014-01-01

    Highlights: • Heats of solution of NaAlO 2 (s) were measured at five temperatures up to 250 °C. • Standard molar enthalpies of solution were determined from the measured heats of solution. • Standard molar enthalpies of solution were correlated with the density model. • The density model allows us to determine the standard molar heat capacities of reaction. - Abstract: Heats of solution of sodium aluminum oxide, NaAlO 2 (s), were measured in aqueous sodium hydroxide solutions using a Tian–Calvet heat-flow calorimeter (Setaram, Model C80) with high pressure “batch cells” made of hastelloy C-276, at five temperatures from (373.15 to 523.15) K, steam saturation pressure, and concentrations from (0.02 to 0.09) mol · kg −1 . Standard molar enthalpies of solution, Δ soln H ∘ , and relative standard molar enthalpies, [H ∘ (T) − H ∘ (298.15 K)], of NaAl(OH) 4 (aq) were determined from the measured heats of solution. The results were fitted with the “density” model. The temperature dependence of Δ soln H ∘ from the model yielded the standard molar heat capacities of reaction, Δ soln C p ∘ , from which standard partial molar heat capacities for aqueous aluminate, C p ∘ [A1(OH) 4 − ,aq], were calculated. Standard partial molar enthalpies of formation, Δ f H ∘ , and entropies, S ∘ , of A1(OH) 4 − (aq) were also determined. The values for C p ∘ [A1(OH) 4 − ,aq] agree with literature data determined up to T = 413 K from enthalpy of solution and heat capacity measurements to within the combined experimental uncertainties. They are consistent with differential heat capacity measurements up to T = 573 K from Schrödle et al. (2010) [29] using the same calorimeter, but this method has the advantage that measurements could be made at much lower concentrations in the presence of an excess concentration of ligand. To our knowledge, these are the first standard partial molar heat capacities measured under hydrothermal conditions by the

  7. The comet assay – from toy to tool

    Directory of Open Access Journals (Sweden)

    Guenter Speit

    2015-04-01

    Full Text Available The comet assay is nowadays the most common method for measuring DNA damage and repair in single cells. It is based on the microelectrophoretic study published by Ostling and Johanson (1984 and was developed by Singh and coworkers (1988 to a versatile technique for quantitation of low levels of DNA damage in individual cells. This alkaline version still is the basis for the triumphant success of the comet assay in basic research into mechanisms of DNA damage and DNA repair, genotoxicity testing, ecotoxicology and human biomonitoring. Important technical improvements (e.g., the use of precoated slides, introduction of image analysis, high throughput methods, automated scoring systems made the assay more robust and more efficient. Modifications of the standard protocol provide more specific information on the type and biological significance of the damage studied. The introduction of lesion-specific endonucleases allowed the characterization of oxidative base damage, alkylation damage and UV-induced pyrimidine dimers. The combination with fluorescence in situ hybridization (FISH made it possible to identify DNA of particular chromosome regions and measure effects of damage and repair in particular genes. The comet assay is being increasingly used in genotoxicity testing. In particular, the in vivo comet assay has become a component of some genotoxicity test strategies and generally accepted test protocols have evolved over the years. A large international collaborative trial sponsored by the Japanese Center for the Validation of Alternative Methods (JaCVAM was recently completed and an OECD test guideline was approved. The comet assay is widely used in human biomonitoring to measure DNA damage as a marker of exposure to genotoxic agents or to investigate genoprotective effects. However, there are still problems in comparing results from different laboratories and there is need for reducing inter-laboratory variation and identification of standard

  8. REFLECTION OF THE SOCIAL STANDARDS AS A CONDITION OF A SUCCESSFUL SOCIALIZATION OF THE CHILDREN WITH A HIGH CREATIVE POTENTIAL

    Directory of Open Access Journals (Sweden)

    Antonina L. Leutina

    2015-01-01

    Full Text Available The aim of this article is to study the problem of reflection of social norms and its impact on the process of socialization of children with a high creative potential.Methods. Processes of socialization and reflection of social norms are considered in this article on the basis of the methodology of the system and mental activity analysis developed by G. P. Schedrovitsky, the founder of the Moscow Methodological Study Group. The article provides the comparative analysis of the adaptation and activity approach to socialization of children with a high creative potential according to such parameters as the object, socialization mechanisms, characteristics of social norms, social controls, personal controls, personal qualities, level of social mobility, and nature of the process of socialization.Results and scientific novelty. The author puts forward the thesis about irrational character and «off-limits» of judgements of the majority of social norms in a modern society, and shows distinction of social and personal regulators of a reflection of these norms. One of the main conclusions is the following: rationalisation and reflection of social norms are the important conditions of social dynamics and social development.It has been found that unconscious personal acceptance of social norms that is typical of the adaptation approach leads to two opposite results: 1 successful adaptation due to uncritical acceptance of social norms by the child, which leads to decrease in the diversity of ways of activity and specificity of its products, and, as a result, failure to realize the creative potential; 2 desocialization of the child in case of keeping the level of creative abilities and the possibility of its realization.The activity approach which is based on reflexive mechanisms represents absolutely different methodological opportunities for solving the problem of socialization of children with a high creative potential. The activity approach contributes

  9. The use of comet assay in plant toxicology: recent advances

    Directory of Open Access Journals (Sweden)

    Conceição LV Santos

    2015-06-01

    Full Text Available The systematic study of genotoxicity in plants induced by contaminants and other stress agents has been hindered to date by the lack of reliable and robust biomarkers. The comet assay is a versatile and sensitive method for the evaluation of DNA damages and DNA repair capacity at single-cell level. Due to its simplicity and sensitivity, and the small number of cells required to obtain robust results, the use of plant comet assay has drastically increased in the last decade. For years its use was restricted to a few model species, e.g. Allium cepa, Nicotiana tabacum, Vicia faba, or Arabidopsis thaliana but this number largely increased in the last years. Plant comet assay has been used to study the genotoxic impact of radiation, chemicals including pesticides, phytocompounds, heavy metals, nanoparticles or contaminated complex matrices. Here we will review the most recent data on the use of this technique as a standard approach for studying the genotoxic effects of different stress conditions on plants. Also, we will discuss the integration of information provided by the comet assay with other DNA-damage indicators, and with cellular responses including oxidative stress, cell division or cell death. Finally, we will focus on putative relations between transcripts related with DNA damage pathways, DNA replication and repair, oxidative stress and cell cycle progression that have been identified in plant cells with comet assays demonstrating DNA damage.

  10. Comparison of Batch Assay and Random Assay Using Automatic Dispenser in Radioimmunoassay

    Energy Technology Data Exchange (ETDEWEB)

    Moon, Seung Hwan; Jang, Su Jin; Kang, Ji Yeon; Lee, Dong Soo; Chung, June Key; Lee, Myung Chul [Seoul Metropolitan Government Seoul National University Boramae Medical Center, Seoul (Korea, Republic of); Lee, Ho Young; Shin, Sun Young; Min, Gyeong Sun; Lee, Hyun Joo [Seoul National University college of Medicine, Seoul (Korea, Republic of)

    2009-08-15

    Radioimmunoassay (RIA) was usually performed by the batch assay. To improve the efficiency of RIA without increase of the cost and time, random assay could be a choice. We investigated the possibility of the random assay using automatic dispenser by assessing the agreement between batch assay and random assay. The experiments were performed with four items; Triiodothyronine (T3), free thyroxine (fT4), Prostate specific antigen (PSA), Carcinoembryonic antigen (CEA). In each item, the sera of twenty patients, the standard, and the control samples were used. The measurements were done 4 times with 3 hour time intervals by random assay and batch assay. The coefficient of variation (CV) of the standard samples and patients' data in T3, fT4, PSA, and CEA were assessed. ICC (Intraclass correlation coefficient) and coefficient of correlation were measured to assessing the agreement between two methods. The CVs (%) of T3, fT4, PSA, and CEA measured by batch assay were 3.2+-1.7%, 3.9+-2.1%, 7.1+-6.2%, 11.2+-7.2%. The CVs by random assay were 2.1+-1.7%, 4.8+-3.1%, 3.6+-4.8%, and 7.4+-6.2%. The ICC between the batch assay and random assay were 0.9968 (T3), 0.9973 (fT4), 0.9996 (PSA), and 0.9901 (CEA). The coefficient of correlation between the batch assay and random assay were 0.9924(T3), 0.9974 (fT4), 0.9994 (PSA), and 0.9989 (CEA) (p<0.05). The results of random assay showed strong agreement with the batch assay in a day. These results suggest that random assay using automatic dispenser could be used in radioimmunoassay

  11. An opto-chemical assay for mold detection in processing tomatoes

    Science.gov (United States)

    Potts, Steven James

    Fungal (mold) contamination is an important indicator of low quality raw product or unsanitary processing conditions in the food industry. A quantitative lectin assay was developed that was less expensive, faster, and more precise than the industry standard Howard mold count. This assay, based on a fluorescent-labeled lectin isolated from wheat germ, has a selective affinity for the chitin in fungal cell walls. Assay values had high linear correlations (from r 2 = .72 to r2 = .99) with fungal biomass for ten fungal species of greatest importance to the California processing tomato industry. One hundred raw tomato juice samples with natural mold infections were collected, as part of the normal California processing tomato inspection program, from commercial processing tomato loads. The raw juice samples were sent to four processor quality control laboratories, where an industry standard Howard mold count was conducted on blind triplicates of each of the samples. The lectin assay was also conducted on blind replicates of the raw juice samples. The correlation between the lectin assay and the Howard mold count (r = .85) was as strong as the correlation between the Howard mold counts for two facilities. The assay had significantly better (alpha = .01) precision than the Howard mold count, with an average coefficient of variation of 8%, compared with 38% for the Howard mold count. The assay is objective, can be conducted in under six minutes, and has the potential to replace mold testing in both raw and processed products.

  12. Determination and Standardization of Analytical Conditions for Dissolved Boron in Coastal Waters of East Sea in Korea by ICP-OES

    Science.gov (United States)

    Yoon, H.; Shin, M.; Yoon, C.; Lee, J.

    2005-12-01

    The analysis of metals in seawaters has been an important subject for many years. Achieving low-level detection limits as well as overcoming high matrix effect are requested in seawater analysis especially elements of interest are present in various chemical forms. Among them, boron is one of the widely distributed elements in nature and its concentrations of about 10 ppm in the Earth's crust and about 4.5 ppm in the seawater as borates. In seawater boron concentration exhibit a linear relationship to the amount of chloride ion present. Boron had been considered as one of the valuable elements to recover from seawaters for commercial use. Currently, we launched research team for the production of valuable metals from seawaters in Korea that can be used commercially. Several metals including boron were already under serious studies. In this study we aim to prepare standardized operational procedures in analysis of boron during pilot study for boron recovery as pilot recovery process. Inductively coupled plasma Optical Emission Spectrometry (ICP-OES) method is preferred for the analysis of the low levels of boron found in environmental samples such as seawater. In order to develop test method for the determination of dissolved Boron from East Sea Seawater in Korea, all soluble boron present in seawater has been tested and accuracy of measurement was checked from the sampling step. The result of analysis of boron in seawaters presents many difficult problems, ionization of from the alkali and alkaline earth metals. And the problems also exist in handling nebulizer and injector tubes in high saline solutions. The scope of this study was to determine boron which can contain up to 35psu dissolved salt. The work also included comparing various analytical methods for better accurate results in several solution conditions. Dilution, standard addition, matrix matching calibration methods was thoroughly tested differently and detailed operating conditions for using auxiliary

  13. Random assay in radioimmunoassay: Feasibility and application compared with batch assay

    International Nuclear Information System (INIS)

    Lee, Jung Min; Lee, Hwan Hee; Park, Sohyun; Kim, Tae Sung; Kim, Seok Ki

    2016-01-01

    The batch assay has been conventionally used for radioimmunoassay (RIA) because of its technical robustness and practical convenience. However, it has limitations in terms of the relative lag of report time due to the necessity of multiple assays in a small number of samples compared with the random assay technique. In this study, we aimed to verify whether the random assay technique can be applied in RIA and is feasible in daily practice. The coefficients of variation (CVs) of eight standard curves within a single kit were calculated in a CA-125 immunoradiometric assay (IRMA) for the reference of the practically ideal CV of the CA-125 kit. Ten standard curves of 10 kits from 2 prospectively collected lots (pLot) and 85 standard curves of 85 kits from 3 retrospectively collected lots (Lot) were obtained. Additionally, the raw measurement data of both 170 control references and 1123 patients' sera were collected retrospectively between December 2015 and January 2016. A standard curve of the first kit of each lot was used as a master standard curve for a random assay. The CVs of inter-kits were analyzed in each lot, respectively. All raw measurements were normalized by decay and radioactivity. The CA-125 values from control samples and patients' sera were compared using the original batch assay and random assay. In standard curve analysis, the CVs of inter-kits in pLots and Lots were comparable to those within a single kit. The CVs from the random assay with normalization were similar to those from the batch assay in the control samples (CVs % of low/high concentration; Lot1 2.71/1.91, Lot2 2.35/1.83, Lot3 2.83/2.08 vs. Lot1 2.05/1.21, Lot2 1.66/1.48, Lot3 2.41/2.14). The ICCs between the batch assay and random assay using patients' sera were satisfactory (Lot1 1.00, Lot2 0.999, Lot3 1.00). The random assay technique could be successfully applied to the conventional CA-125 IRMA kits. The random assay showed strong agreement with the batch assay. The

  14. Random assay in radioimmunoassay: Feasibility and application compared with batch assay

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Jung Min; Lee, Hwan Hee; Park, Sohyun; Kim, Tae Sung; Kim, Seok Ki [Dept. of Nuclear MedicineNational Cancer Center, Goyang (Korea, Republic of)

    2016-12-15

    The batch assay has been conventionally used for radioimmunoassay (RIA) because of its technical robustness and practical convenience. However, it has limitations in terms of the relative lag of report time due to the necessity of multiple assays in a small number of samples compared with the random assay technique. In this study, we aimed to verify whether the random assay technique can be applied in RIA and is feasible in daily practice. The coefficients of variation (CVs) of eight standard curves within a single kit were calculated in a CA-125 immunoradiometric assay (IRMA) for the reference of the practically ideal CV of the CA-125 kit. Ten standard curves of 10 kits from 2 prospectively collected lots (pLot) and 85 standard curves of 85 kits from 3 retrospectively collected lots (Lot) were obtained. Additionally, the raw measurement data of both 170 control references and 1123 patients' sera were collected retrospectively between December 2015 and January 2016. A standard curve of the first kit of each lot was used as a master standard curve for a random assay. The CVs of inter-kits were analyzed in each lot, respectively. All raw measurements were normalized by decay and radioactivity. The CA-125 values from control samples and patients' sera were compared using the original batch assay and random assay. In standard curve analysis, the CVs of inter-kits in pLots and Lots were comparable to those within a single kit. The CVs from the random assay with normalization were similar to those from the batch assay in the control samples (CVs % of low/high concentration; Lot1 2.71/1.91, Lot2 2.35/1.83, Lot3 2.83/2.08 vs. Lot1 2.05/1.21, Lot2 1.66/1.48, Lot3 2.41/2.14). The ICCs between the batch assay and random assay using patients' sera were satisfactory (Lot1 1.00, Lot2 0.999, Lot3 1.00). The random assay technique could be successfully applied to the conventional CA-125 IRMA kits. The random assay showed strong agreement with the batch assay. The

  15. Microbead agglutination based assays

    KAUST Repository

    Kodzius, Rimantas

    2013-01-21

    We report a simple and rapid room temperature assay for point-of-care (POC) testing that is based on specific agglutination. Agglutination tests are based on aggregation of microbeads in the presence of a specific analyte thus enabling the macroscopic observation. Such tests are most often used to explore antibody-antigen reactions. Agglutination has been used for protein assays using a biotin/streptavidin system as well as a hybridization based assay. The agglutination systems are prone to selftermination of the linking analyte, prone to active site saturation and loss of agglomeration at high analyte concentrations. We investigated the molecular target/ligand interaction, explaining the common agglutination problems related to analyte self-termination, linkage of the analyte to the same bead instead of different microbeads. We classified the agglutination process into three kinds of assays: a two- component assay, a three-component assay and a stepped three- component assay. Although we compared these three kinds of assays for recognizing DNA and protein molecules, the assay can be used for virtually any molecule, including ions and metabolites. In total, the optimized assay permits detecting analytes with high sensitivity in a short time, 5 min, at room temperature. Such a system is appropriate for POC testing.

  16. Radiometric assays for glycerol, glucose, and glycogen

    International Nuclear Information System (INIS)

    Bradley, D.C.; Kaslow, H.R.

    1989-01-01

    We have developed radiometric assays for small quantities of glycerol, glucose and glycogen, based on a technique described by Thorner and Paulus for the measurement of glycerokinase activity. In the glycerol assay, glycerol is phosphorylated with [32P]ATP and glycerokinase, residual [32P]ATP is hydrolyzed by heating in acid, and free [32P]phosphate is removed by precipitation with ammonium molybdate and triethylamine. Standard dose-response curves were linear from 50 to 3000 pmol glycerol with less than 3% SD in triplicate measurements. Of the substances tested for interference, only dihydroxyacetone gave a slight false positive signal at high concentration. When used to measure glycerol concentrations in serum and in media from incubated adipose tissue, the radiometric glycerol assay correlated well with a commonly used spectrophotometric assay. The radiometric glucose assay is similar to the glycerol assay, except that glucokinase is used instead of glycerokinase. Dose response was linear from 5 to 3000 pmol glucose with less than 3% SD in triplicate measurements. Glucosamine and N-acetylglucosamine gave false positive signals when equimolar to glucose. When glucose concentrations in serum were measured, the radiometric glucose assay agreed well with hexokinase/glucose-6-phosphate dehydrogenase (H/GDH)-based and glucose oxidase/H2O2-based glucose assays. The radiometric method for glycogen measurement incorporates previously described isolation and digestion techniques, followed by the radiometric assay of free glucose. When used to measure glycogen in mouse epididymal fat pads, the radiometric glycogen assay correlated well with the H/GDH-based glycogen assay. All three radiometric assays offer several practical advantages over spectral assays

  17. Results of in vitro chemosensitivity assays

    International Nuclear Information System (INIS)

    Tanigawa, Nobuhiko; Morimoto, Hideki; Akita, Toshiaki; Inoue, Hiroshi; Tanaka, Takeo.

    1986-01-01

    The authors reviewed their experiences to date with chemosensitivity testing of 629 tumors by human tumor clonogenic assay (HTCA) and of 199 tumors by scintillation assay (SA). HTCA and SA were both performed using a double-layer-soft-agar system with continuous exposure of cells to one concentration of standard anticancer drugs. Overall, 60 % of specimens in HTCA and 58 % in SA produced significant growth in vitro. HTCA was 52 % (13/25) reliable for predicting in vivo sensitivity, and 95 % (36/38) reliable for in vivo resistance, whereas SA was 40 % (8/20) reliable for in vivo sensitivity and 88 % (21/24) for in vivo resistance. In vitro success rates were variable, depending on the tumor histology. In vitro growth of gastric cancer specimens was characteristically lower than that of colon cancer specimens (48 % and 60 % in HTCA, and 46 % and 68 % in SA, respectively). (p < 0.005). Optimal in vitro-in vivo drug concentrations and culture conditions are still being defined. Correlation studies of in vitro-in vivo responses of gastrointestinal cancers suggested that in vitro concentrations of 5-fluorouracil and mitomycin C used in this study were considerably higher than their optimal doses. Tumor cell heterogeneity poses significant problems in the clinical use of chemosensitivity assays. In this last study, we sought evidence of tumor heterogeneity by comparing chemosensitivity responses between : 1) different portions of a single tumor, 2) a primary and a metastatic biopsy taken from a patient on the same day, and 3) different metastases from a patient taken on the same day. The results demonstrated the presence of considerable heterogeneity of response to chemotherapy among different tumors from the same patient, and even within the same tumor. The reported discrepancies of in vitro and in vivo sensitivity may be due to such therapeutic heterogeneity among tumors. (J.P.N.)

  18. Problems with the PTH assays.

    Science.gov (United States)

    Cavalier, Etienne; Delanaye, Pierre; Nyssen, Laurent; Souberbielle, Jean-Claude

    2015-05-01

    Even if the first assay for parathyroid hormone (PTH) was published in the early 1960s, its determination remains a challenge even today. Indeed, in the circulation, PTH is present in its active form (PTH 1-84), but many PTH fragments can also be present. These fragments accumulate when renal function declines and are recognized, at different extents, by the 2nd generation ("intact") PTH assays that are widely used in the clinical laboratories. Some assays, called "3rd generation PTH" do not recognize these fragments, but are not available everywhere. Hence, different problems are also linked with PTH determination. Among them, one can cite the lack of a reference method, the lack of standardization of the assays and, sometimes, the lack of consistent reference range. We can also point out stability problems and a large intra-individual variation. A workgroup is working on these problems under the auspices of the IFCC and we hope that some of these problems will be resolved in the next years. In this article, we will discuss all the possible issues of PTH determination. Copyright © 2015. Published by Elsevier Masson SAS.

  19. Automation of the dicentric chromosome assay and related assays

    International Nuclear Information System (INIS)

    Balajee, Adayabalam S.; Dainiak, Nicholas

    2016-01-01

    Dicentric Chromosome Assay (DCA) is considered to be the 'gold standard' for personalized dose assessment in humans after accidental or incidental radiation exposure. Although this technique is superior to other cytogenetic assays in terms of specificity and sensitivity, its potential application to radiation mass casualty scenarios is highly restricted because DCA is time consuming and labor intensive when performed manually. Therefore, it is imperative to develop high throughput automation techniques to make DCA suitable for radiological triage scenarios. At the Cytogenetic Biodosimetry Laboratory in Oak Ridge, efforts are underway to develop high throughput automation of DCA. Current status on development of various automated cytogenetic techniques in meeting the biodosimetry needs of radiological/nuclear incident(s) will be discussed

  20. Absolute nuclear material assay

    Science.gov (United States)

    Prasad, Manoj K [Pleasanton, CA; Snyderman, Neal J [Berkeley, CA; Rowland, Mark S [Alamo, CA

    2010-07-13

    A method of absolute nuclear material assay of an unknown source comprising counting neutrons from the unknown source and providing an absolute nuclear material assay utilizing a model to optimally compare to the measured count distributions. In one embodiment, the step of providing an absolute nuclear material assay comprises utilizing a random sampling of analytically computed fission chain distributions to generate a continuous time-evolving sequence of event-counts by spreading the fission chain distribution in time.

  1. Absolute nuclear material assay

    Science.gov (United States)

    Prasad, Manoj K [Pleasanton, CA; Snyderman, Neal J [Berkeley, CA; Rowland, Mark S [Alamo, CA

    2012-05-15

    A method of absolute nuclear material assay of an unknown source comprising counting neutrons from the unknown source and providing an absolute nuclear material assay utilizing a model to optimally compare to the measured count distributions. In one embodiment, the step of providing an absolute nuclear material assay comprises utilizing a random sampling of analytically computed fission chain distributions to generate a continuous time-evolving sequence of event-counts by spreading the fission chain distribution in time.

  2. Tricyclic antidepressant radioreceptor assay

    International Nuclear Information System (INIS)

    Innis, R.B.; Tune, L.; Rock, R.; Depaulo, R.; U'Prichard, D.C.; Snyder, S.M.

    1979-01-01

    A receptor assay for tricyclic antidepressants described here is based on the ability of these drugs to compete with [ 3 H]-3-guinuclidnyl benzilate ( 3 H-QNB) for binding to muscarinic cholinergic receptors in rat brain membranes. The assay is sensitive, in that it can detect, for example, 2ng/ml nortriptyline in plasma. Seven plasma samples from depressed patients treated with nortriptyline were assayed with the radioreceptor and gas liquid chromatographic methods, and the results from these two methods were almost identical. This assay should be used cautiously, if at all, in patients treated with other drugs that have potent anticholinergic effects. (Auth.)

  3. 21 CFR 864.7415 - Abnormal hemoglobin assay.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Abnormal hemoglobin assay. 864.7415 Section 864... hemoglobin assay. (a) Identification. An abnormal hemoglobin assay is a device consisting of the reagents... hemoglobin types. (b) Classification. Class II (performance standards). [45 FR 60618, Sept. 12, 1980] ...

  4. Endogenous Locus Reporter Assays.

    Science.gov (United States)

    Liu, Yaping; Hermes, Jeffrey; Li, Jing; Tudor, Matthew

    2018-01-01

    Reporter gene assays are widely used in high-throughput screening (HTS) to identify compounds that modulate gene expression. Traditionally a reporter gene assay is built by cloning an endogenous promoter sequence or synthetic response elements in the regulatory region of a reporter gene to monitor transcriptional activity of a specific biological process (exogenous reporter assay). In contrast, an endogenous locus reporter has a reporter gene inserted in the endogenous gene locus that allows the reporter gene to be expressed under the control of the same regulatory elements as the endogenous gene, thus more accurately reflecting the changes seen in the regulation of the actual gene. In this chapter, we introduce some of the considerations behind building a reporter gene assay for high-throughput compound screening and describe the methods we have utilized to establish 1536-well format endogenous locus reporter and exogenous reporter assays for the screening of compounds that modulate Myc pathway activity.

  5. Micropatterned comet assay enables high throughput and sensitive DNA damage quantification

    Science.gov (United States)

    Ge, Jing; Chow, Danielle N.; Fessler, Jessica L.; Weingeist, David M.; Wood, David K.; Engelward, Bevin P.

    2015-01-01

    The single cell gel electrophoresis assay, also known as the comet assay, is a versatile method for measuring many classes of DNA damage, including base damage, abasic sites, single strand breaks and double strand breaks. However, limited throughput and difficulties with reproducibility have limited its utility, particularly for clinical and epidemiological studies. To address these limitations, we created a microarray comet assay. The use of a micrometer scale array of cells increases the number of analysable comets per square centimetre and enables automated imaging and analysis. In addition, the platform is compatible with standard 24- and 96-well plate formats. Here, we have assessed the consistency and sensitivity of the microarray comet assay. We showed that the linear detection range for H2O2-induced DNA damage in human lymphoblastoid cells is between 30 and 100 μM, and that within this range, inter-sample coefficient of variance was between 5 and 10%. Importantly, only 20 comets were required to detect a statistically significant induction of DNA damage for doses within the linear range. We also evaluated sample-to-sample and experiment-to-experiment variation and found that for both conditions, the coefficient of variation was lower than what has been reported for the traditional comet assay. Finally, we also show that the assay can be performed using a 4× objective (rather than the standard 10× objective for the traditional assay). This adjustment combined with the microarray format makes it possible to capture more than 50 analysable comets in a single image, which can then be automatically analysed using in-house software. Overall, throughput is increased more than 100-fold compared to the traditional assay. Together, the results presented here demonstrate key advances in comet assay technology that improve the throughput, sensitivity, and robustness, thus enabling larger scale clinical and epidemiological studies. PMID:25527723

  6. Solid phase assays

    International Nuclear Information System (INIS)

    Reese, M.G.; Johnson, L.R.; Ransom, D.K.

    1980-01-01

    In a solid phase assay for quantitative determination of biological and other analytes, a sample such as serum is contacted with a receptor for the analyte being assayed, the receptor being supported on a solid support. No tracer for the analyte is added to the sample before contacting with the receptor; instead the tracer is contacted with the receptor after unbound analyte has been removed from the receptor. The assay can be otherwise performed in a conventional manner but can give greater sensitivity. (author)

  7. Clinical effectiveness and cost-effectiveness of use of therapeutic monitoring of tumour necrosis factor alpha (TNF-α) inhibitors [LISA-TRACKER® enzyme-linked immunosorbent assay (ELISA) kits, TNF-α-Blocker ELISA kits and Promonitor® ELISA kits] versus standard care in patients with Crohn's disease: systematic reviews and economic modelling.

    Science.gov (United States)

    Freeman, Karoline; Connock, Martin; Auguste, Peter; Taylor-Phillips, Sian; Mistry, Hema; Shyangdan, Deepson; Court, Rachel; Arasaradnam, Ramesh; Sutcliffe, Paul; Clarke, Aileen

    2016-11-01

    Systematic reviews and economic modelling of clinical effectiveness and cost-effectiveness of therapeutic monitoring of tumour necrosis factor alpha (TNF-α) inhibitors [using LISA-TRACKER ® enzyme-linked immunosorbent assay (ELISA) kits (Theradiag, Marne La Vallee, France, or Alpha Laboratories, Heriot, UK), TNF-α-Blocker ELISA kits (Immundiagnostik AG, Bensheim, Germany) and Promonitor ® ELISA kits (Proteomika, Progenika Biopharma, Bizkaia, Spain)] versus standard care for Crohn's disease (CD). Multiple electronic databases were searched from inception to December 2014 in order to identify primary studies and meta-analyses. Patients with moderate to severe active CD treated with infliximab (IFX) (Remicade ® , Merck Sharp & Dohme Ltd, Kenilworth, NJ, USA) or adalimumab (ADA) (Humira ® , AbbVie Inc., North Chicago, IL, USA). Monitoring of serum anti-TNF-α (IFX or ADA) and/or of anti-drug antibody levels using test assays with a test-treatment algorithm. Standard care. Any patient-related outcome, test agreement and cost-effectiveness estimates. The quality assessments used recognised checklists (Quality Assessment of Diagnostic Accuracy Studies-2, Cochrane, Philips and Consolidated Health Economic Evaluation Reporting Standards). Evidence was synthesised using narrative review and meta-analysis. A Markov model was built in TreeAge Pro 2013 (TreeAge Software, Inc., Williamstown, MA, USA). The model had a 4-week cycle and a 10-year time horizon, adopted a NHS and Personal Social Services perspective and used a linked evidence approach. Costs were adjusted to 2013/14 prices and discounted at 3.5%. We included 68 out of 2434 and 4 out of 2466 studies for the clinical effectiveness and cost-effectiveness reviews, respectively. Twenty-three studies comparing test methods were identified. Evidence on test concordance was sparse and contradictory, offering scant data for a linked evidence approach. Three studies [two randomised controlled trials (RCTs) and one

  8. Assay of ribulose bisphosphate carboxylase

    International Nuclear Information System (INIS)

    Pike, C.; Berry, J.

    1987-01-01

    Assays of ribulose bisphosphate carboxylase (rubisco) can be used to illustrate many properties of photosynthetic systems. Many different leaves have been assayed with this standard procedure. The tissue is ground with a mortar and pestle in extraction buffer. The supernatant after centrifugation is used as the source of enzyme. Buffer, RuBP, [ 14 C]-NaHCO 3 , and enzyme are combined in a scintillation vial; the reaction is run for 1 min at 30 0 . The acid-stable products are counted. Reproducibility in student experiments has been excellent. The assay data can be combined with analyses of leaf properties such as fresh and dry weight, chlorophyll and protein content, etc. Students have done projects such as the response of enzyme to temperature and to various inhibitors. They also report on the use of a transition state analog, carboxyarabinitol bisphosphate, to titrate the molar concentration of rubisco molecules (active sites) in an enzyme sample. Thus, using crude extracts the catalytic activity of a sample can be compared to the absolute quantity of enzyme or to the turnover number

  9. Low- and high-order accurate boundary conditions: From Stokes to Darcy porous flow modeled with standard and improved Brinkman lattice Boltzmann schemes

    Energy Technology Data Exchange (ETDEWEB)

    Silva, Goncalo, E-mail: goncalo.nuno.silva@gmail.com [Irstea, Antony Regional Centre, HBAN, 1 rue Pierre-Gilles de Gennes CS 10030, 92761 Antony cedex (France); Talon, Laurent, E-mail: talon@fast.u-psud.fr [CNRS (UMR 7608), Laboratoire FAST, Batiment 502, Campus University, 91405 Orsay (France); Ginzburg, Irina, E-mail: irina.ginzburg@irstea.fr [Irstea, Antony Regional Centre, HBAN, 1 rue Pierre-Gilles de Gennes CS 10030, 92761 Antony cedex (France)

    2017-04-15

    The present contribution focuses on the accuracy of reflection-type boundary conditions in the Stokes–Brinkman–Darcy modeling of porous flows solved with the lattice Boltzmann method (LBM), which we operate with the two-relaxation-time (TRT) collision and the Brinkman-force based scheme (BF), called BF-TRT scheme. In parallel, we compare it with the Stokes–Brinkman–Darcy linear finite element method (FEM) where the Dirichlet boundary conditions are enforced on grid vertices. In bulk, both BF-TRT and FEM share the same defect: in their discretization a correction to the modeled Brinkman equation appears, given by the discrete Laplacian of the velocity-proportional resistance force. This correction modifies the effective Brinkman viscosity, playing a crucial role in the triggering of spurious oscillations in the bulk solution. While the exact form of this defect is available in lattice-aligned, straight or diagonal, flows; in arbitrary flow/lattice orientations its approximation is constructed. At boundaries, we verify that such a Brinkman viscosity correction has an even more harmful impact. Already at the first order, it shifts the location of the no-slip wall condition supported by traditional LBM boundary schemes, such as the bounce-back rule. For that reason, this work develops a new class of boundary schemes to prescribe the Dirichlet velocity condition at an arbitrary wall/boundary-node distance and that supports a higher order accuracy in the accommodation of the TRT-Brinkman solutions. For their modeling, we consider the standard BF scheme and its improved version, called IBF; this latter is generalized in this work to suppress or to reduce the viscosity correction in arbitrarily oriented flows. Our framework extends the one- and two-point families of linear and parabolic link-wise boundary schemes, respectively called B-LI and B-MLI, which avoid the interference of the Brinkman viscosity correction in their closure relations. The performance of LBM

  10. Lateral flow assays.

    Science.gov (United States)

    Koczula, Katarzyna M; Gallotta, Andrea

    2016-06-30

    Lateral flow assays (LFAs) are the technology behind low-cost, simple, rapid and portable detection devices popular in biomedicine, agriculture, food and environmental sciences. This review presents an overview of the principle of the method and the critical components of the assay, focusing on lateral flow immunoassays. This type of assay has recently attracted considerable interest because of its potential to provide instantaneous diagnosis directly to patients. The range and interpretation of results and parameters used for evaluation of the assay will also be discussed. The main advantages and disadvantages of LFAs will be summarized and relevant future improvements to testing devices and strategies will be proposed. Finally, the major recent advances and future diagnostic applications in the LFA field will be explored. © 2016 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

  11. Lateral flow assays

    Science.gov (United States)

    Koczula, Katarzyna M.

    2016-01-01

    Lateral flow assays (LFAs) are the technology behind low-cost, simple, rapid and portable detection devices popular in biomedicine, agriculture, food and environmental sciences. This review presents an overview of the principle of the method and the critical components of the assay, focusing on lateral flow immunoassays. This type of assay has recently attracted considerable interest because of its potential to provide instantaneous diagnosis directly to patients. The range and interpretation of results and parameters used for evaluation of the assay will also be discussed. The main advantages and disadvantages of LFAs will be summarized and relevant future improvements to testing devices and strategies will be proposed. Finally, the major recent advances and future diagnostic applications in the LFA field will be explored. PMID:27365041

  12. Proceedings of the CSNI workshop on International Standard Problem 48 - Analysis of 1:4-scale prestressed concrete containment vessel model under severe accident conditions

    International Nuclear Information System (INIS)

    2005-01-01

    At the CSNI meeting in June 2002, the proposal for an International Standard Problem on containment integrity (ISP 48) based on the NRC/NUPEC/Sandia test was approved. Objectives were to extend the understanding of capacities of actual containment structures based on results of the recent PCCV Model test and other previous research. The ISP was sponsored by the USNRC, and results had been made available thanks to NUPEC and to the USNRC. Sandia National Laboratory was contracted to manage the technical aspects of the ISP. At the end of the ISP48, a workshop was organized in Lyon, France on April 6-7, 2005 hosted by Electricite de France. Its overall objective was to present results obtained by participants in the ISP 48 and to assess the current practices and the state of the art with respect to the calculation of concrete structures under severe accident conditions. Experience from other areas in civil engineering related to the modelling of complex structures was greatly beneficial to all. Information obtained as a result of this assessment were utilized to develop a consensus on these calculations and identify issues or 'gaps' in the present knowledge for the primary purpose of formulating and prioritizing research needs on this topic. The ISP48 exercise was published in the report referenced NEA/CSNI/R(2005)5 in 3 volumes. Volume 1 contains the synthesis of the exercise; Volumes 2 and 3 contain individual contributions of participating organizations. The CSNI Working Group on the Integrity and Ageing and in particular its sub-group on the behaviour of concrete structures has produced extensive material over the last few years. The complete list of references is given in this document. These proceedings gather the papers and presentations given by the participants at the Lyon workshop

  13. Dose-response evaluation of the standardized ileal digestible tryptophan : lysine ratio to maximize growth performance of growing-finishing gilts under commercial conditions.

    Science.gov (United States)

    Gonçalves, M A D; Tokach, M D; Bello, N M; Touchette, K J; Goodband, R D; DeRouchey, J M; Woodworth, J C; Dritz, S S

    2017-11-16

    Environmental regulations as well as economic incentives have resulted in greater use of synthetic amino acids in swine diets. Tryptophan is typically the second limiting amino acid in corn-soybean meal-based diets. However, using corn-based co-products emphasizes the need to evaluate the pig's response to increasing Trp concentrations. Therefore, the objective of these studies was to evaluate the dose-response to increasing standardized ileal digestible (SID) Trp : Lys on growth performance of growing-finishing gilts housed under large-scale commercial conditions. Dietary treatments consisted of SID Trp : Lys of 14.5%, 16.5%, 18.0%, 19.5%, 21.0%, 22.5% and 24.5%. The study was conducted in four experiments of 21 days of duration each, and used corn-soybean meal-based diets with 30% distillers dried grains with solubles. A total of 1166, 1099, 1132 and 975 gilts (PIC 337×1050, initially 29.9±2.0 kg, 55.5±4.8 kg, 71.2±3.4 kg and 106.2±3.1 kg BW, mean±SD) were used. Within each experiment, pens of gilts were blocked by BW and assigned to one of the seven dietary treatments and six pens per treatment with 20 to 28 gilts/pen. First, generalized linear mixed models were fit to data from each experiment to characterize performance. Next, data were modeled across experiments and fit competing dose-response linear and non-linear models and estimate SID Trp : Lys break points or maximums for performance. Competing models included broken-line linear (BLL), broken-line quadratic and quadratic polynomial (QP). For average daily gain (ADG), increasing the SID Trp : Lys increased growth rate in a quadratic manner (Pgilts ranged from a minimum of 16.9% for maximum G : F to 23.5% for maximum ADG.

  14. Quantification of particle sizes with metal replication under standard freeze-etching conditions: a gold ball standard for calibrating shadow widths was used to measure freeze-etched globular proteins.

    Science.gov (United States)

    Ruben, G C

    1995-11-01

    The real size of platinum-carbon (Pt-C) replicated particles is not directly equivalent to either its metal-coated diameter or its shadow width. This paper describes two indirect methods, shadow widths and coated particle diameters, for determining a particle's actual size beneath a Pt-C replication film. Both produce equivalent measurements using the same standardized conditions: 2.3 nm Pt-C films deposited at a 45 degree angle on an approximately -100 degrees C surface in a 10(-6) torr vacuum. For the first method, gold balls nucleated in a partial pressure of helium and deposited on flat indirect carbon films (root mean square roughness of 0.8 nm) on 400 mesh grids were used as test particles for calibrating shadow widths as a function of particle size. The gold ball test specimens were replicated, and a distribution of Pt-C shadow widths orthogonal to the Pt-C deposition direction was measured and averaged for gold balls 1.5 +/- 0.25 nm, 2.0 +/- 0.25 nm, etc. The diameter of each gold ball was measured within the Pt-C film along with its shadow width because the Pt-C did not obscure or adhere well to the gold. The shadow width distributions for each gold size do not differ significantly from log normal. Two proteins, the lactose repressor and the mitochondrial ATPase, F1, were also used as replication test objects. Negative staining of both proteins was conducted to measure their average diameters. In the second method, a distribution of Pt-C-coated lac repressor diameters perpendicular to the shadow direction was measured. The Pt-C film thickness measured on the quartz crystal monitor was subtracted from the average metal-coated protein diameter to obtain the lac repressor's diameter. The Pt-C-coated particle diameter distributions also did not differ significantly from log normal. While doing this work it was discovered that outgassing the Pt-C electron gun greatly affected Pt-C film granularity: 19 sec produced a high contrast, granular Pt-C film, whereas

  15. Comparison of in-house biotin-avidin tetanus IgG enzyme-linked-immunosorbent assay (ELISA) with gold standard in vivo mouse neutralization test for the detection of low level antibodies.

    Science.gov (United States)

    Sonmez, Cemile; Coplu, Nilay; Gozalan, Aysegul; Akin, Lutfu; Esen, Berrin

    2017-06-01

    Detection of anti-tetanus antibody levels is necessary for both determination of the immune status of individuals and also for planning preventive measures. ELISA is the preferred test among in vitro tests however it can be affected by the cross reacting antibodies. A previously developed in-house ELISA test was found not reliable for the antibody levels ≤1.0IU/ml. A new method was developed to detect low antibody levels correctly. The aim of the present study was to compare the results of the newly developed in-house biotin-avidin tetanus IgG ELISA test with the in vivo mouse neutralization test, for the antibody levels ≤1.0IU/ml. A total of 54 serum samples with the antibody levels of three different levels, =0.01IU/ml, 0.01-0.1IU/ml, 0.1-1IU/ml, which were detected by in vivo mouse neutralization test were studied by the newly developed in-house biotin-avidin tetanus IgG ELISA test. Test was validated by using five different concentrations (0.01IU/ml, 0.06IU/ml, 0.2IU/ml, 0.5IU/ml, 1.0IU/ml). A statistically significant correlation (r 2 =0.9967 p=0,001) between in vivo mouse neutralization test and in-house biotin-avidin tetanus IgG ELISA test, was observed. For the tested concentrations intra-assay, inter-assay, accuracy, sensitivity, specificity and coefficients of variations were determined as ≤15%. In-house biotin-avidin tetanus IgG ELISA test can be an alternative method to in vivo mouse neutralization method for the detection of levels ≤1.0IU/ml. By using in-house biotin-avidin tetanus IgG ELISA test, individuals with non protective levels, will be reliably detected. Copyright © 2017. Published by Elsevier B.V.

  16. Calibration method for a radwaste assay system

    International Nuclear Information System (INIS)

    Dulama, C.; Dobrin, R.; Toma, Al.; Paunoiu, C.

    2004-01-01

    A waste assay system entirely designed and manufactured in the Institute for Nuclear Research is used in radwaste treatment and conditioning stream to ensure compliance with national repository radiological requirements. Usually, waste assay systems are calibrated by using various experimental arrangements including calibration phantoms. The paper presents a comparative study concerning the efficiency calibration performed by shell source method and a semiempirical, computational method based on a Monte Carlo algorithm. (authors)

  17. Towards an international standard for detection and typing botulinum neurotoxin-producing Clostridia types A, B, E and F in food, feed and environmental samples: a European ring trial study to evaluate a real-time PCR assay.

    Science.gov (United States)

    Fenicia, Lucia; Fach, Patrick; van Rotterdam, Bart J; Anniballi, Fabrizio; Segerman, Bo; Auricchio, Bruna; Delibato, Elisabetta; Hamidjaja, Raditijo A; Wielinga, Peter R; Woudstra, Cedric; Agren, Joakim; De Medici, Dario; Knutsson, Rickard

    2011-03-01

    A real-time PCR method for detection and typing of BoNT-producing Clostridia types A, B, E, and F was developed on the framework of the European Research Project "Biotracer". A primary evaluation was carried out using 104 strains and 17 clinical and food samples linked to botulism cases. Results showed 100% relative accuracy, 100% relative sensitivity, 100% relative specificity, and 100% selectivity (inclusivity on 73 strains and exclusivity on 31 strains) of the real-time PCR against the reference cultural method combined with the standard mouse bioassay. Furthermore, a ring trial study performed at four different European laboratories in Italy, France, the Netherlands, and Sweden was carried out using 47 strains, and 30 clinical and food samples linked to botulism cases. Results showed a concordance of 95.7% among the four laboratories. The reproducibility generated a relative standard deviation in the range of 2.18% to 13.61%. Considering the high level of agreement achieved between the laboratories, this real-time PCR is a suitable method for rapid detection and typing of BoNT-producing Clostridia in clinical, food and environmental samples and thus support the use of it as an international standard method. Copyright © 2011 Elsevier B.V. All rights reserved.

  18. Ultra fast and sensitive liquid chromatography tandem mass spectrometry based assay for galactose-1-phosphate uridylyltransferase and galactokinase deficiencies.

    Science.gov (United States)

    Li, Yijun; Ptolemy, Adam S; Harmonay, Lauren; Kellogg, Mark; Berry, Gerard T

    2011-01-01

    The diagnosis of transferase and galactokinase deficiency galactosemia usually involves the measurement of erythrocyte galactose-1-phosphate uridylyltransferase (GALT) and galactokinase (GALK) enzyme activity, respectively. The current gold standard assays for these enzymes are radioactive assays, which are laborious and/or incapable of measuring low enzyme activities. To further our knowledge of genotype-phenotype relationships, we had developed an assay for GALT activity alone using LC-MS/MS. In this study we generated a robust and sensitive LC-MS/MS based GALT and GALK assay using a novel normal phase chromatographic condition. We improved upon our earlier assay by drastically reducing the instrument run time and eliminating the use of an ion pairing reagent. Stable isotope labeled substrates were utilized in the GALT and GALK assays. The enzymatic products ([(13)C(6)]-uridine diphosphate galactose in GALT assay and [(13)C(6)]-galactose-1-phosphate in GALK assay) were quantified in a 3 min LC-MS/MS run. The assays were sensitive enough to allow for the quantification of enzyme activities as low as 0.2% and 0.3% of normal control values in the GALT and GALK assays, respectively. Thirty-three samples from non-galactosemic patients were assayed to have erythrocyte GALT activity of 23.4±4.2 and GALK activity of 1.8±0.47 (mean±SD) μmol⋅(g Hgb)(-1) h(-1). Erythrocyte GALT activities in a cohort of 16 patients with classic or severe galactosemia were measured: 4 patients had GALT activity less than 1% of normal control values and the remaining 12 had no detectable GALT activity. No GALK activity was detected in a GALK deficient sample we analyzed. Lastly, we tested the feasibility of adapting this LC-MS/MS based GALT/GALK assay as a newborn screening (NBS) test. Copyright © 2010 Elsevier Inc. All rights reserved.

  19. Characterization of XR-RV3 GafChromic{sup ®} films in standard laboratory and in clinical conditions and means to evaluate uncertainties and reduce errors

    Energy Technology Data Exchange (ETDEWEB)

    Farah, J., E-mail: jad.farah@irsn.fr; Clairand, I.; Huet, C. [External Dosimetry Department, Institut de Radioprotection et de Sûreté Nucléaire (IRSN), BP-17, 92260 Fontenay-aux-Roses (France); Trianni, A. [Medical Physics Department, Udine University Hospital S. Maria della Misericordia (AOUD), p.le S. Maria della Misericordia, 15, 33100 Udine (Italy); Ciraj-Bjelac, O. [Vinca Institute of Nuclear Sciences (VINCA), P.O. Box 522, 11001 Belgrade (Serbia); De Angelis, C. [Department of Technology and Health, Istituto Superiore di Sanità (ISS), Viale Regina Elena 299, 00161 Rome (Italy); Delle Canne, S. [Fatebenefratelli San Giovanni Calibita Hospital (FBF), UOC Medical Physics - Isola Tiberina, 00186 Rome (Italy); Hadid, L.; Waryn, M. J. [Radiology Department, Hôpital Jean Verdier (HJV), Avenue du 14 Juillet, 93140 Bondy Cedex (France); Jarvinen, H.; Siiskonen, T. [Radiation and Nuclear Safety Authority (STUK), P.O. Box 14, 00881 Helsinki (Finland); Negri, A. [Veneto Institute of Oncology (IOV), Via Gattamelata 64, 35124 Padova (Italy); Novák, L. [National Radiation Protection Institute (NRPI), Bartoškova 28, 140 00 Prague 4 (Czech Republic); Pinto, M. [Istituto Nazionale di Metrologia delle Radiazioni Ionizzanti (ENEA-INMRI), C.R. Casaccia, Via Anguillarese 301, I-00123 Santa Maria di Galeria (RM) (Italy); Knežević, Ž. [Ruđer Bošković Institute (RBI), Bijenička c. 54, 10000 Zagreb (Croatia)

    2015-07-15

    Purpose: To investigate the optimal use of XR-RV3 GafChromic{sup ®} films to assess patient skin dose in interventional radiology while addressing the means to reduce uncertainties in dose assessment. Methods: XR-Type R GafChromic films have been shown to represent the most efficient and suitable solution to determine patient skin dose in interventional procedures. As film dosimetry can be associated with high uncertainty, this paper presents the EURADOS WG 12 initiative to carry out a comprehensive study of film characteristics with a multisite approach. The considered sources of uncertainties include scanner, film, and fitting-related errors. The work focused on studying film behavior with clinical high-dose-rate pulsed beams (previously unavailable in the literature) together with reference standard laboratory beams. Results: First, the performance analysis of six different scanner models has shown that scan uniformity perpendicular to the lamp motion axis and that long term stability are the main sources of scanner-related uncertainties. These could induce errors of up to 7% on the film readings unless regularly checked and corrected. Typically, scan uniformity correction matrices and reading normalization to the scanner-specific and daily background reading should be done. In addition, the analysis on multiple film batches has shown that XR-RV3 films have generally good uniformity within one batch (<1.5%), require 24 h to stabilize after the irradiation and their response is roughly independent of dose rate (<5%). However, XR-RV3 films showed large variations (up to 15%) with radiation quality both in standard laboratory and in clinical conditions. As such, and prior to conducting patient skin dose measurements, it is mandatory to choose the appropriate calibration beam quality depending on the characteristics of the x-ray systems that will be used clinically. In addition, yellow side film irradiations should be preferentially used since they showed a lower

  20. Evaluation of the Diagnostic Accuracy of a Typhoid IgM Flow Assay for the Diagnosis of Typhoid Fever in Cambodian Children Using a Bayesian Latent Class Model Assuming an Imperfect Gold Standard

    Science.gov (United States)

    Moore, Catrin E.; Pan-Ngum, Wirichada; Wijedoru, Lalith P. M.; Sona, Soeng; Nga, Tran Vu Thieu; Duy, Pham Thanh; Vinh, Phat Voong; Chheng, Kheng; Kumar, Varun; Emary, Kate; Carter, Michael; White, Lisa; Baker, Stephen; Day, Nicholas P. J.; Parry, Christopher M.

    2014-01-01

    Rapid diagnostic tests are needed for typhoid fever (TF) diagnosis in febrile children in endemic areas. Five hundred children admitted to the hospital in Cambodia between 2009 and 2010 with documented fever (≥ 38°C) were investigated using blood cultures (BCs), Salmonella Typhi/Paratyphi A real-time polymerase chain reactions (PCRs), and a Typhoid immunoglobulin M flow assay (IgMFA). Test performance was determined by conventional methods and Bayesian latent class modeling. There were 32 cases of TF (10 BC- and PCR-positive cases, 14 BC-positive and PCR-negative cases, and 8 BC-negative and PCR-positive cases). IgMFA sensitivity was 59.4% (95% confidence interval = 41–76), and specificity was 97.8% (95% confidence interval = 96–99). The model estimate sensitivity for BC was 81.0% (95% credible interval = 54–99). The model estimate sensitivity for PCR was 37.8% (95% credible interval = 26–55), with a specificity of 98.2% (95% credible interval = 97–99). The model estimate sensitivity for IgMFA (≥ 2+) was 77.9% (95% credible interval = 58–90), with a specificity of 97.5% (95% credible interval = 95–100). The model estimates of IgMFA sensitivity and specificity were comparable with BCs and better than estimates using conventional analysis. PMID:24218407

  1. Assay method and compositions

    International Nuclear Information System (INIS)

    1977-01-01

    Methods are described for measuring catecholamine levels in human and animal body fluids and tissues using the catechol-O-methyl-transferase (COMT) radioassay. The assay involves incubating the biological sample with COMT and the tritiated methyl donor, S-adenosyl-L-methionine( 3 H)-methyl. The O-methylated ( 3 H) epinephrine and/or norepinephrine are extracted and oxidised to vanillin- 3 H which in turn is extracted and its radioactivity counted. When analysing dopamine levels the assay is extended by vanillin- 3 H and raising the pH of the aqueous periodate phase from which O-methylated ( 3 H) dopamine is extracted and counted. The assay may be modified depending on whether measurements of undifferentiated total endogenous catecholamine levels or differential analyses of the catecholamine levels are being performed. The sensitivity of the assay can be as low as 5 picograms for norepinephrine and epinephrine and 12 picograms for dopamine. The assemblance of the essential components of the assay into a kit for use in laboratories is also described. (U.K.)

  2. Radioreceptor assay for insulin

    International Nuclear Information System (INIS)

    Suzuki, Kazuo

    1975-01-01

    Radioreceptor assay of insulin was discussed from the aspects of the measuring method, its merits and problems to be solved, and its clinical application. Rat liver 10 x g pellet was used as receptor site, and enzymatic degradation of insulin by the system contained in this fraction was inhibited by adding 1 mM p-CMB. 125 I-labelled porcine insulin was made by lactoperoxidase method under overnight incubation at 4 0 C and later purification by Sephadex G-25 column and Whatman CF-11 cellulose powder. Dog pancreatic vein serum insulin during and after the glucose load was determined by radioreceptor assay and radioimmunoassay resulting that both measurements accorded considerably. Radioreceptor assay would clarify the pathology of disorders of glucose metabolism including diabetes. (Tsukamoto, Y.)

  3. Realization of the developing potential of training to computer science in conditions of adoption of the second generation state educational standards

    Directory of Open Access Journals (Sweden)

    Сергей Георгиевич Григорьев

    2010-03-01

    Full Text Available In article requirements to training to computer science and an information technology, formulated with a position of planned results presented in the standard of the second generation are described.

  4. Standardization and optimization of core sampling procedure for carbon isotope analysis in eucalyptus and variation in carbon isotope ratios across species and growth conditions

    CSIR Research Space (South Africa)

    Raju, M

    2011-11-01

    Full Text Available 13C is a well established surrogate for water use efficiency (WUE). However, variation due to aspect, length of branch and position In canopy can cause potential errors. Hence, experiments were conducted to standardize the sampling procedures...

  5. Radioreceptor assay for GH

    International Nuclear Information System (INIS)

    Tsushima, Toshio; Matsuzaki, Fukashi

    1975-01-01

    Radioreceptor assay (RRA) of growth hormone (GH) was studied using the protein which specifically bound to GH presenting in the liver of rabbits. 100,000g pellet of the liver homogenate was used as receptor source. The factors which affected the results of RRA such as salt, temperature and incubation time, were discussed. As same as in other RRA methods, serum protein inhibited non-specifically 125 I-GH binding in this method. In this assay, serum GH less than 5ng/ml could not be detected. The difference between the value obtained by RRA and that by radioimmunoassay was compared with reference to the patients with acromegalia. (Tsukamoto, Y.)

  6. Lateral flow strip assay

    Science.gov (United States)

    Miles, Robin R [Danville, CA; Benett, William J [Livermore, CA; Coleman, Matthew A [Oakland, CA; Pearson, Francesca S [Livermore, CA; Nasarabadi, Shanavaz L [Livermore, CA

    2011-03-08

    A lateral flow strip assay apparatus comprising a housing; a lateral flow strip in the housing, the lateral flow strip having a receiving portion; a sample collection unit; and a reagent reservoir. Saliva and/or buccal cells are collected from an individual using the sample collection unit. The sample collection unit is immersed in the reagent reservoir. The tip of the lateral flow strip is immersed in the reservoir and the reagent/sample mixture wicks up into the lateral flow strip to perform the assay.

  7. Lateral flow assays

    NARCIS (Netherlands)

    Posthuma-Trumpie, G.A.; Amerongen, van A.

    2012-01-01

    A simple version of immunochemical-based methods is the Lateral Flow Assay (LFA). It is a dry chemistry technique (reagents are included); the fluid from the sample runs through a porous membrane (often nitrocellulose) by capillary force. Typically the membrane is cut as a strip of 0.5*5 cm. In most

  8. Trueness verification of actual creatinine assays in the European market demonstrates a disappointing variability that needs substantial improvement. An international study in the framework of the EC4 creatinine standardization working group.

    Science.gov (United States)

    Delanghe, Joris R; Cobbaert, Christa; Galteau, Marie-Madeleine; Harmoinen, Aimo; Jansen, Rob; Kruse, Rolf; Laitinen, Päivi; Thienpont, Linda M; Wuyts, Birgitte; Weykamp, Cas; Panteghini, Mauro

    2008-01-01

    The European In Vitro Diagnostics (IVD) directive requires traceability to reference methods and materials of analytes. It is a task of the profession to verify the trueness of results and IVD compatibility. The results of a trueness verification study by the European Communities Confederation of Clinical Chemistry (EC4) working group on creatinine standardization are described, in which 189 European laboratories analyzed serum creatinine in a commutable serum-based material, using analytical systems from seven companies. Values were targeted using isotope dilution gas chromatography/mass spectrometry. Results were tested on their compliance to a set of three criteria: trueness, i.e., no significant bias relative to the target value, between-laboratory variation and within-laboratory variation relative to the maximum allowable error. For the lower and intermediate level, values differed significantly from the target value in the Jaffe and the dry chemistry methods. At the high level, dry chemistry yielded higher results. Between-laboratory coefficients of variation ranged from 4.37% to 8.74%. Total error budget was mainly consumed by the bias. Non-compensated Jaffe methods largely exceeded the total error budget. Best results were obtained for the enzymatic method. The dry chemistry method consumed a large part of its error budget due to calibration bias. Despite the European IVD directive and the growing needs for creatinine standardization, an unacceptable inter-laboratory variation was observed, which was mainly due to calibration differences. The calibration variation has major clinical consequences, in particular in pediatrics, where reference ranges for serum and plasma creatinine are low, and in the estimation of glomerular filtration rate.

  9. DNA-based mutation assay GPMA (genome profiling-based mutation assay): reproducibility, parts-per-billion scale sensitivity, and introduction of a mammalian-cell-based approach.

    Science.gov (United States)

    Kumari, Parmila; Gautam, Sunita Ghimire; Baba, Misato; Tsukiashi, Motoki; Matsuoka, Koji; Yasukawa, Kiyoshi; Nishigaki, Koichi

    2017-12-01

    Genome profiling-based mutation assay (GPMA) is, to date, the only DNA sequence-based mutation assay that directly measures DNA alterations induced by mutagens. Here, the all-important congruence of mutagen assignment between DNA-based GPMA and the phenotype-based Ames test (the gold standard of mutagen assays) was confirmed qualitatively and semi-quantitatively by means of 94 chemical species (including previously examined 64). The high sensitivity (on the order of 10 ppb) and reproducibility of GPMA were also corroborated by the match between virtually independent experiments conducted in the distant past (10 years ago) and recently. Meanwhile, a standard experimental framework was established: the conditions of 100 parts per billion (ppb) concentration of a chemical and 15-generation culture of Escherichia coli. Moreover, a mammalian cell line (NIH 3T3) was shown to be suitable as a tester organism for the GPMA approach. Preliminary experimental results suggested that this approach can provide a qualitatively equivalent and quantitatively different mutagen assay results relative to the bacteria-based GPMA (renamed as bGPMA). This finding confirmed the effectiveness of the GPMA approach and indicates that mGPMA is a promising way to detect mammalian-cell mutagens. © The Authors 2017. Published by Oxford University Press on behalf of the Japanese Biochemical Society. All rights reserved.

  10. Reporter Gene Assays in Ecotoxicology.

    Science.gov (United States)

    Elad, Tal; Belkin, Shimshon

    The need for simple and rapid means for evaluating the potential toxic effects of environmental samples has prompted the development of reporter gene assays, based on tester cells (bioreporters) genetically engineered to report on sample toxicity by producing a readily quantifiable signal. Bacteria are especially suitable to serve as bioreporters owing to their fast responses, low cost, convenient preservation, ease of handling, and amenability to genetic manipulations. Various bacterial bioreporters have been introduced for general toxicity and genotoxicity assessment, and the monitoring of endocrine disrupting and dioxin-like compounds has been mostly covered by similarly engineered eukaryotic cells. Some reporter gene assays have been validated, standardized, and accredited, and many others are under constant development. Efforts are aimed at broadening detection spectra, lowering detection thresholds, and combining toxicity identification capabilities with characterization of the toxic effects. Taking advantage of bacterial robustness, attempts are also being made to incorporate bacterial bioreporters into field instrumentation for online continuous monitoring or on-site spot checks. However, key hurdles concerning test validation, cell preservation, and regulatory issues related to the use of genetically modified organisms still remain to be overcome.

  11. Standardization of biodosimetry operations

    International Nuclear Information System (INIS)

    Dainiak, Nicholas

    2016-01-01

    Methods and procedures for generating, interpreting and scoring the frequency of dicentric chromosomes vary among cytogenetic biodosimetry laboratories (CBLs). This variation adds to the already considerable lack of precision inherent in the dicentric chromosome assay (DCA). Although variability in sample collection, cell preparation, equipment and dicentric frequency scoring can never be eliminated with certainty, it can be substantially minimized, resulting in reduced scatter and improved precision. Use of standard operating procedures and technician exchange may help to mitigate variation. Although the development and adoption of international standards (ISO 21243 and ISO 19238) has helped to reduce variation in standard operating procedures (SOPs), all CBLs must maintain process improvement, and those with challenges may require additional assistance. Sources of variation that may not be readily apparent in the SOPs for sample collection and processing include variability in ambient laboratory conditions, media, serum lot and quantity and the use of particular combinations of cytokines. Variability in maintenance and calibration of metafer equipment, and in scoring criteria, reader proficiency and personal factors may need to be addressed. The calibration curve itself is a source of variation that requires control, using the same known-dose samples among CBLs, measurement of central tendency, and generation of common curves with periodic reassessment to detect drifts in dicentric yield. Finally, the dose estimate should be based on common scoring criteria, using of the z-statistic. Although theoretically possible, it is practically impossible to propagate uncertainty over the entire calibration curve due to the many factors contributing to variance. Periodic re-evaluation of the curve is needed by comparison with newly published curves (using statistical analysis of differences) and determining their potential causes. (author)

  12. Dual isotope assays

    International Nuclear Information System (INIS)

    Smith, G.F.W.; Stevens, R.A.J.; Jacoby, B.

    1980-01-01

    Dual isotope assays for thyroid function are performed by carrying out a radio-immunoassay for two of thyroxine (T4), tri-iodothyronine (T3), thyroid stimulating hormone (TSH), and thyroxine binding globulin (TBG), by a method wherein a version of one of the thyroid components, preferably T4 or T3 is labelled with Selenium-75 and the version of the other thyroid component is labelled with a different radionuclide, preferably Iodine-125. (author)

  13. Cytotoxicity assay automation

    Science.gov (United States)

    Levinthal, E. C.; Payne, R. O.

    1971-01-01

    The design and construction of a system to automatically test HLP antigens are described. Major efforts were made to test and evaluate the performance of such a system, and compare its performance with nonautomatic tissue typing techniques. The system is based on the fluorochromatic cytotoxicity assay. Results show the system will work but is subject to malfunctions after a few samplings, and poses problems in showing correctly the necessary readings.

  14. Performance of monitors, radiodiagnostic level, in standard X-ray beams conditions; Comportamento de monitores, nivel radiodiagnostico, em feixes padronizados de radiacao X

    Energy Technology Data Exchange (ETDEWEB)

    Potiens, Maria da Penha A.; Caldas, Linda V.E. [Instituto de Pesquisas Energeticas e Nucleares (IPEN), Sao Paulo, SP (Brazil)

    2002-07-01

    (The Calibration Laboratory of IPEN implanted the radiation qualities recommended by the IEC 1267 standard, in the X radiation Neo-Diagnomax system for the calibration of instruments used in diagnostic radiology. Since 1997 this service has been offered to the users. Sixteen different models of ionization chambers used in measurements in diagnostic radiology have been calibrated since then. The nine more utilized models of ionization chambers (volumes varying from 3 to 1800 cm{sup 3}), calibrated more than one time during the period 1997-2001, were selected for analysis. The results show that only three ionization chambers presented the energy dependence higher than 5%, that is the recommended value by the international standards for this kind of instrument. (author)

  15. On the Training in the Conditions of Application of Professional Standards in the Field of Education (on the Example of Educational Psychologist

    Directory of Open Access Journals (Sweden)

    Egorova M.A.

    2017-11-01

    Full Text Available The article presents a modern approach to the design of the basic educational program for the training of a pedagogue-psychologist, the purpose and objectives of which are set by the professional standard “Pedagogue-psychologist (educational psychologist”. The structure of the basic educational program of bachelor and master's levels is designed according to the modular principle, and the key mechanism for achieving educational results - professional competences - is networking with partner organizations (schools, kindergartens, psychological and medico-social centers. The main criteria of network partners for the training of psychologists are described. The training module is characterized as a logically completed, relatively independent part of the educational program that forms a certain competence or group of competencies. In the curriculum the module acts as an educational unit uniting theory, practice, research work and the final modular attestation of the student. The advantages of the modular structure of the educational program, coupled with the professional standard, are presented.

  16. HBA1c: clinical and biological agreement for standardization of assay methods. Report by the experts of ALFEDIAM (Association de Langue Française pour lEtude du Diabète et des Maladies Métabolique) and SFBC (Société Française de Biologie Clinique).

    Science.gov (United States)

    Gillery, P; Bordas-Fonfrède, M; Chapelle, J P; Drouin, P; Hue, G; Lévy-Marchal, C; Périer, C; Sélam, J L; Slama, G; Thivolet, C; Vialettes, B

    1999-09-01

    Glycohaemoglobin, and particularly haemoglobin A1c(HbA1c), assays have been used for many years to retrospectively evaluate the glycaemic control of diabetic patients. Cut-off values have been established for deciding treatment modifications. The techniques used in the laboratories however exhibit varying quality, and all of them are not yet standardized. The consequence is an under-utilization of this test, especially in non-hospital practice. In this context, working groups of Société Française de Biologie Clinique (SFBC), Association de Langue Française pour l'Etude du Diabète et des Maladies Métaboliques (ALFEDIAM) and Société Française d'Endocrinologie (SFE) have met together, in order to analyze the national status, and to propose practical recommendations for implementing a standardization process on the basis of international experiences. It is recommended to exclusively express results as HbA1c percentage, using methods standardized and certified by comparison to reference methods such as those using Diabetes Control and Complications Trial (DCCT) values. Simultaneously, contacts have been established with manufacturers, and the realisation of periodic quality control surveys was encouraged.

  17. Colorimetric micro-assay for accelerated screening of mould inhibitors

    Science.gov (United States)

    Carol A. Clausen; Vina W. Yang

    2013-01-01

    Since current standard laboratory methods are time-consuming macro-assays that rely on subjective visual ratings of mould growth, rapid and quantitative laboratory methods are needed to screen potential mould inhibitors for use in and on cellulose-based products. A colorimetric micro-assay has been developed that uses XTT tetrazolium salt to enzymatically assess...

  18. Study of radiation conditions for obtaining standard beam computed tomography; Estudo das condicoes de radiacao para obtencao do feixe padrao em tomografia computadorizada

    Energy Technology Data Exchange (ETDEWEB)

    Andrade, Lucio das Chagas de; Peixoto, Jose Guilherme Pereira, E-mail: lucio-andrade@hotmail.com [Instituto de Radioprotecao e Dosimetria (IRD/CNEN-RJ), Rio de Janeiro, RJ (Brazil)

    2015-07-01

    In Brazil there is a need to expand the traceability for the calibration of cameras used in dosimetry in CT. Thus, in order to promote the expansion of the radiation conditions RQT, LNMRI disseminate this greatness. These radiation conditions may be characterized in terms of tension in the X-ray tube (PPV), the first half-value layer (1{sup st} HVL) and homogeneity coefficient (CH). The LNMRI achieved satisfactory results within the international specifications suggested by IEC 61267 and TRS 457. (author)

  19. Methodology for determination of plasma cortisol in fish using Competitive Enzyme-Linked Immunosorbent Assay (ELISA)

    DEFF Research Database (Denmark)

    Velasco-Santamaría, Yohana M.; Cruz-Casallas, Pablo E.

    2007-01-01

    Objective. To determine plasma cortisol procedure in fish using competitive enzymelinked immunosorbent assay (ELISA). Materials and methods. Two plasma samples of juveniles rainbow trout Oncorhynchus mykiss were analized by using ELISA human kit for cortisol assay. For standard curve calibration ...

  20. Radon assay for SNO+

    Energy Technology Data Exchange (ETDEWEB)

    Rumleskie, Janet [Laurentian University, Greater Sudbury, Ontario (Canada)

    2015-12-31

    The SNO+ experiment will study neutrinos while located 6,800 feet below the surface of the earth at SNOLAB. Though shielded from surface backgrounds, emanation of radon radioisotopes from the surrounding rock leads to back-grounds. The characteristic decay of radon and its daughters allows for an alpha detection technique to count the amount of Rn-222 atoms collected. Traps can collect Rn-222 from various positions and materials, including an assay skid that will collect Rn-222 from the organic liquid scintillator used to detect interactions within SNO+.

  1. Radiorespirometic assay device

    International Nuclear Information System (INIS)

    Levin, G.V.; Straat, P.A.

    1981-01-01

    A radiorespirometic assay device is described in which the presence of microorganisms in a sample is determined by placing the sample in contact with a metabolisable radioactive labelled substrate, collecting any gas evolved, exposing a photosensitive material to the gas and determining if a spot is produced on the material. A spot indicates the presence of radioactivity showing that the substrate has been metabolized by a microorganism. Bacteria may be detected in body fluids, hospital operating rooms, water, food, cosmetics and drugs. (U.K.)

  2. Design the Professional Educational Environment for Teacher in the Conditions of the Introduction and Development of Professional Standard of the Activity: the Leningrad Oblast Experience

    Directory of Open Access Journals (Sweden)

    Bayeva I.A.,

    2016-07-01

    Full Text Available On the example of the educational system of Leningrad oblast the authors analyse how the environmental approach can be used in the implementation of the professional standard for teachers. The issues concerning the process of the implementation and acquisition of the professional standard are discussed in relation to the innovative project “Safe Educational Environment” carried out in collaboration with the Russian Academy of Education. This project is aimed at setting up the most socially significant components of professional and educational environment as well as at creating and testing a system of psychological safety support for subjects of the educational process in the region. The authors emphasize the advantages of the system approach that implies the introduction of various innovations in education and helps to achieve a synergistic effect in the teacher’s professional activity. Also, they show how the methodology of the environmental approach may be employed in the designing of professional educational environment as one of the technologies in the teacher’s practice.

  3. Standardizing the experimental conditions for using urine in NMR-based metabolomic studies with a particular focus on diagnostic studies: a review

    KAUST Repository

    Emwas, Abdul-Hamid M.

    2014-11-21

    The metabolic composition of human biofluids can provide important diagnostic and prognostic information. Among the biofluids most commonly analyzed in metabolomic studies, urine appears to be particularly useful. It is abundant, readily available, easily stored and can be collected by simple, noninvasive techniques. Moreover, given its chemical complexity, urine is particularly rich in potential disease biomarkers. This makes it an ideal biofluid for detecting or monitoring disease processes. Among the metabolomic tools available for urine analysis, NMR spectroscopy has proven to be particularly well-suited, because the technique is highly reproducible and requires minimal sample handling. As it permits the identification and quantification of a wide range of compounds, independent of their chemical properties, NMR spectroscopy has been frequently used to detect or discover disease fingerprints and biomarkers in urine. Although protocols for NMR data acquisition and processing have been standardized, no consensus on protocols for urine sample selection, collection, storage and preparation in NMR-based metabolomic studies have been developed. This lack of consensus may be leading to spurious biomarkers being reported and may account for a general lack of reproducibility between laboratories. Here, we review a large number of published studies on NMR-based urine metabolic profiling with the aim of identifying key variables that may affect the results of metabolomics studies. From this survey, we identify a number of issues that require either standardization or careful accounting in experimental design and provide some recommendations for urine collection, sample preparation and data acquisition.

  4. Harmonization of immune biomarker assays for clinical studies.

    Science.gov (United States)

    van der Burg, Sjoerd H; Kalos, Michael; Gouttefangeas, Cécile; Janetzki, Sylvia; Ottensmeier, Christian; Welters, Marij J P; Romero, Pedro; Britten, Cedrik M; Hoos, Axel

    2011-11-09

    Assays that measure a patient's immune response play an increasingly important role in the development of immunotherapies. The inherent complexity of these assays and independent protocol development between laboratories result in high data variability and poor reproducibility. Quality control through harmonization--based on integration of laboratory-specific protocols with standard operating procedures and assay performance benchmarks--is one way to overcome these limitations. Harmonization guidelines can be widely implemented to address assay performance variables. This process enables objective interpretation and comparison of data across clinical trial sites and also facilitates the identification of relevant immune biomarkers, guiding the development of new therapies.

  5. Beyond Renewable Portfolio Standards: An Assessment of Regional Supply and Demand Conditions Affecting the Future of Renewable Energy in the West; Report and Executive Summary

    Energy Technology Data Exchange (ETDEWEB)

    Hurlbut, D. J.; McLaren, J.; Gelman, R.

    2013-08-01

    This study assesses the outlook for utility-scale renewable energy development in the West once states have met their renewable portfolio standard (RPS) requirements. In the West, the last state RPS culminates in 2025, so the analysis uses 2025 as a transition point on the timeline of RE development. Most western states appear to be on track to meet their final requirements, relying primarily on renewable resources located relatively close to the customers being served. What happens next depends on several factors including trends in the supply and price of natural gas, greenhouse gas and other environmental regulations, consumer preferences, technological breakthroughs, and future public policies and regulations. Changes in any one of these factors could make future renewable energy options more or less attractive.

  6. Present conditions and perspectives of the standardization of the energy efficiency in Mexico; Actualidad y perspectivas del programa de normalizacion de la eficiencia energetica en Mexico

    Energy Technology Data Exchange (ETDEWEB)

    Chu Pulido, Henry Anli; Sanchez Ramos, Itha [Instituto de Investigaciones Electricas, Cuernavaca (Mexico)

    1998-12-31

    The first energy efficiency standards of compulsory type were implemented in some states of the U.S. and in 1990 this country`s Federal government adopted them, since then, they have spread all over the world. This paper describes that under the frame of the National Program for Energy Modernization (1900-1994) and the National Development Program (1995-2000) of Mexico were considered as a priority the rational use of the energy, the conservation of the energy resources and the environmental protection. It is particularly mentioned, the standardization program of energy efficiency that has been developed that includes the limiting specification of the energy consumption or equipment efficiency, an essay method to determine the energy performance and the description of the label that will inform the users of the equipment energy performance [Espanol] Las primeras normas de eficiencia energetica de tipo obligatorio se implantaron en algunos estados de Estados Unidos y en 1990 las adopto el gobierno federal de ese pais, a partir de entonces se han difundido por todo el mundo. En el presente trabajo se describe que en el marco del Programa Nacional de Modernizacion Energetica (1900-1994) y del Plan Nacional de Desarrollo (1995-2000) de Mexico, se consideran como prioridad el uso racional de energia, la conservacion de recursos energeticos y la proteccion al medio ambiente. Se menciona particularmente, el programa de normalizacion de eficiencia energetica que se ha desarrollado incluye la especificacion del limite de consumo energetico o eficiencia del equipo, un metodo de ensayo para determinar el rendimiento energetico y la descripcion de la etiqueta que informara a los usuarios sobre el comportamiento energetico del equipo

  7. A new fluorescent assay for enalapril maleate.

    Science.gov (United States)

    de los A Oliva, María; Sombra, Lorena L; Olsina, Roberto A; Masi, Adriana N

    2005-09-01

    A new spectrofluorimetric method for the enalapril maleate monitoring was studied. Enalapril maleate was found to be highly photolabile. This drug was evaluated according to photodegradation assay at pH 2.5 and 6. Enalapril maleate was exposed to UVA-UVB radiations. Under these specific conditions was found as degradation product, the diketopiperazine. The modification of the fluorescent properties of enalapril maleate in solution after exposure UV-radiation and the degradation mechanisms were studied. The photodegradation was followed by the developed spectrofluorimetric assay.

  8. Symbiotic properties of Bradyrhizobium sp. (Lupinus assayed on serradella plants

    Directory of Open Access Journals (Sweden)

    Mieczysława Deryło

    2014-01-01

    Full Text Available Physiological and symbiotic properties of Bradyrhizobium sp. (Lupinus nodule isolates were compared to the standard slow-growing Bradyrhizobium sp. (Lupinus strain USDA 3045. Lupine nodules isolates showed typical characteristics for bradyrhizobial strains and nodulated small seed legume, serradella (Ornithopus sativus, in tube test. We observed a permanent physiological segregation of the effective (Fix' and ineffective (Fix- symbiotic phenotype for all tested bradyrhizobial strains during the growth of serradella in plant tube test. The ultrastructural differences between Fix* and Fix serradella nodules were observed. Rapid and visible nodulation as well as easy assay of the reduction of acetylene make serradella a convenient system for studies of Bradyrhizobium sp. (Lupinus strains in laboratory conditions.

  9. Disagreement between Human Papillomavirus Assays

    DEFF Research Database (Denmark)

    Rebolj, Matejka; Preisler, Sarah; Ejegod, Ditte Møller

    2014-01-01

    -65 years (n = 2,881), 23% tested positive on at least one assay, and 42 to 58% of these showed positive agreement on any compared pair of the assays. While 4% of primary screening samples showed abnormal cytology, 6 to 10% were discordant on any pair of assays. A literature review corroborated our findings...

  10. Evaluation of lot-to-lot repeatability and effect of assay media choice in the recombinant Factor C assay.

    Science.gov (United States)

    McKenzie, Jennifer Helen; Alwis, K Udeni; Sordillo, Joanne E; Kalluri, Kesava Srinivas; Milton, Donald Kirby

    2011-06-01

    Measurement of environmental endotoxin exposures is complicated by variability encountered using current biological assay methods arising in part from lot-to-lot variability of the Limulus-amebocyte lysate (LAL) reagents. Therefore, we investigated the lot-to-lot repeatability of commercially available recombinant Factor C (rFC) kits as an alternative to LAL. Specifically, we compared endotoxin estimates obtained from rFC assay of twenty indoor dust samples, using four different extraction and assay media, to endotoxin estimates previously obtained by Limulus amebocyte lysate (LAL) assay and amounts of 3-hydroxy fatty acids (3-OHFA) in lipopolysaccharide (LPS) using gas-chromatography mass spectroscopy (GC-MS). We found that lot-to-lot variability of the rFC assay kits does not significantly alter endotoxin estimates in house dust samples when performed using three of the four assay media tested and that choice of assay media significantly altered endotoxin estimates obtained by rFC assay of house dust samples. Our findings demonstrate lot-to-lot reproducibility of rFC assay of environmental samples and suggest that use of rFC assay performed with Tris buffer or water as the extraction and assay medium for measurement of endotoxin in dust samples may be a suitable choice for developing a standardized methodology.

  11. Principles of validation of diagnostic assays for infectious diseases

    International Nuclear Information System (INIS)

    Jacobson, R.H.

    1998-01-01

    Assay validation requires a series of inter-related processes. Assay validation is an experimental process: reagents and protocols are optimized by experimentation to detect the analyte with accuracy and precision. Assay validation is a relative process: its diagnostic sensitivity and diagnostic specificity are calculated relative to test results obtained from reference animal populations of known infection/exposure status. Assay validation is a conditional process: classification of animals in the target population as infected or uninfected is conditional upon how well the reference animal population used to validate the assay represents the target population; accurate predictions of the infection status of animals from test results (PV+ and PV-) are conditional upon the estimated prevalence of disease/infection in the target population. Assay validation is an incremental process: confidence in the validity of an assay increases over time when use confirms that it is robust as demonstrated by accurate and precise results; the assay may also achieve increasing levels of validity as it is upgraded and extended by adding reference populations of known infection status. Assay validation is a continuous process: the assay remains valid only insofar as it continues to provide accurate and precise results as proven through statistical verification. Therefore, the work required for validation of diagnostic assays for infectious diseases does not end with a time-limited series of experiments based on a few reference samples rather, to assure valid test results from an assay requires constant vigilance and maintenance of the assay, along with reassessment of its performance characteristics for each unique population of animals to which it is applied. (author)

  12. Standardization of RAPD assay for genetic analysis of olive

    African Journals Online (AJOL)

    PRECIOUS

    2009-12-15

    Dec 15, 2009 ... europaea L. Genetic variability and molecular cultivar identification. Genet. Res. Crop Evol. 54(1): 117-128. Mir Ali N, Nabulsi I (2003). Genetic diversity of almond (Prunus dulcis) using RAPD technique, Sci. Hort. 98: 461-471. Nei M (1972). Nei's Original Measures of Genetic Identity and Genetic. Distance.

  13. Standardization of RAPD assay for genetic analysis of olive | Sesli ...

    African Journals Online (AJOL)

    ... also PCR cycles I and II; however, favorable results were attained by PCR Mix III and. PCR cycle III. Evaluable bands were obtained for defining the olive samples by using primers from OP-I. Thus, it was concluded that RAPD profiles are effective in the study of genetic similarities and distances of wild and cultivated olives ...

  14. An acoustic prion assay

    Directory of Open Access Journals (Sweden)

    Gordon Hayward

    2016-12-01

    Full Text Available An acoustic prion assay has been demonstrated for sheep brain samples. Only five false positives and no false negatives were observed in a test of 45 positive and 45 negative samples. The acoustic prion sensor was constructed using a thickness shear mode quartz resonator coated with a covalently bound recombinant prion protein. The characteristic indicator of a scrapie infected sheep brain sample was an observed shoulder in the frequency decrease in response to a sample.The response of the sensor aligns with a conformational shift in the surface protein and with the propagation mechanism of the disease. This alignment is evident in the response timing and shape, dependence on concentration, cross species behaviour and impact of blood plasma. This alignment is far from sufficient to prove the mechanism of the sensor but it does offer the possibility of a rapid and inexpensive additional tool to explore prion disease. Keywords: Prions, Thickness shear mode quartz sensor

  15. Assay of oestrogen

    International Nuclear Information System (INIS)

    Edwards, J.C.

    1981-01-01

    A particular problem with the direct radioimmunoassay of unconjugated oestriol in pregnancy is caused by the increased amount of steroid-binding proteins present in pregnancy serum and plasma. The steroid-binding proteins react with oestriol and 125 I-labelled oestriol during the assay procedure and the steroid-protein bound 125 I-labelled oestriol is precipitated along with the antibody-bound 125 I-labelled oestriol by the ammonium sulphate solution separation system. A novel method is described whereby progesterone (1-20 μg/ml) is used to block the action of steroid-binding proteins in pregnancy serum and plasma samples, thus minimizing interference in a direct radioimmunoassay for unconjugated oestriol using a specific anti-oestriol serum. (U.K.)

  16. TRAINING FUTURE TEACHERS OF COMPUTER SCIENCE FOR WORKING OUT TECHNOLOGICAL CARDS OF LESSONS IN THE CONDITIONS OF REALIZATION OF THE FEDERAL STATE EDUCATIONAL STANDARD FOR GENERAL EDUCATION

    Directory of Open Access Journals (Sweden)

    Екатерина Николаевна Кувшинова

    2017-12-01

    Full Text Available This article is devoted to a problem of readiness of future teachers of informatics for development of flow charts of the lessons displaying the main requirements of Federal state educational standards of the main general education (FGOS of Ltd company to planning and the organization of educational process taking into account system and activity approach in training. Content of system and activity approach in training, the universal educational actions (UEA reveals. Main units of the flow chart of a lesson of informatics are considered. The substantial block of the flow chart of a lesson of informatics determined by a training material which provides achievement of the planned subject results of training, and also forming and development of UUD, all-educational skills, ICT competences, competences of educational and research and project activities is stated.Subject results of training to which the abilities specific to a subject, types of activity on receipt of new knowledge within a subject, to its transformation and application in educational, educational and project and social and project situations, forming of scientific type of thinking, scientific ideas of key theories, types and types of the relations, ownership of scientific terminology, key concepts, methods and acceptances belong [10] are analyzed.Step-by-step training of future teachers of informatics for development of flow charts of lessons is discussed.

  17. Coagulation assays and anticoagulant monitoring.

    Science.gov (United States)

    Funk, Dorothy M Adcock

    2012-01-01

    Anticoagulant therapy, including conventional agents and a variety of new oral, fast-acting drugs, is prescribed for millions of patients annually. Each anticoagulant varies in its effect on routine and specialty coagulation assays and each drug may require distinct laboratory assay(s) to measure drug concentration or activity. This review provides an overview of the assorted assays that can measure anticoagulant drug concentration or activity and includes key assay interferences. The effect of these conventional and new anticoagulant agents on specialty coagulation assays used to evaluate for bleeding or clotting disorders, and whether this impact is physiological or factitious, is included. Also provided is a short review of superwarfarin poisoning and features distinguishing this from warfarin overdose. Knowledge of clinically significant pearls and pitfalls pertinent to coagulation assays in relation to anticoagulant therapy are important to optimize patient care.

  18. Immune chromatography: a quantitative radioimmunological assay

    International Nuclear Information System (INIS)

    Davis, J.W.; Demetriades, M.; Bowen, J.M.

    1984-01-01

    Immune chromatography, a radioimmunological binding assay, employs paper chromatography to separate immune complexes from free antigen and antibodies. During chromatography free antigen and antibodies become distributed throughout the paper, while immune complexes remain near the bottoms of the strips. The chromatographic differences can be made quantitative by using either iodinated antigens or antibodies. Under these conditions nanogram quantities of antigen can be detected or antibodies in sera diluted several 1000-fold. The immune chromatography assay can also be performed as an indirect assay, since the paper strips are cut from nitrocellulose paper. In this case the immune components are absorbed by the paper during chromatography. Antigen is then detected with an iodinated second antibody. The indirect immune chromatography assay is particularly useful for identifying different sera that react with the same antigen. Reaction with the first serum before chromatography reduces the amount of antigen available to the second serum following chromatography. In addition to characterizing the immune chromatography procedure, we discuss the possible applications of chromatography assays for the quantitation of other types of molecular binding interactions. (Auth.)

  19. Automated amperometric plutonium assay system

    International Nuclear Information System (INIS)

    Burt, M.C.

    1985-01-01

    The amperometric titration for plutonium assay has been used in the nuclear industry for over twenty years and has been in routine use at the Hanford Engineering Development Laboratory since 1976 for the analysis of plutonium oxide and mixed oxide fuel material for the Fast Flux Test Facility. It has proven itself to be an accurate and reliable method. The method may be used as a direct end point titration or an excess of titrant may be added and a back titration performed to aid in determination of the end point. Due to the slowness of the PuVI-FeII reaction it is difficult to recognize when the end point is being approached and is very time consuming if the current is allowed to decay to the residual value after each titrant addition. For this reason the back titration in which the rapid FeII-CrVI reaction occurs is used by most laboratories. The back titration is performed by the addition of excess ferrous solution followed by two measured aliquots of standard dichromate with measurement of cell current after each addition

  20. IgG subclasses quantitation: Analytical performance of The Binding Site SPAPLUS® human assay and comparison with Siemens BNII® assay.

    Science.gov (United States)

    Sarnago, Ana; Pascual, Rosa M; Moreno, María J; Laíz, Begoña; Fuster, Oscar

    2018-01-01

    Accurate evaluation of analyzers is highly recommended before these devices are broadly introduced for routine testing. Concerning quantification of IgG subclasses (IgGSc), standardization has not yet been reached and thus different assays might lead to different results. Here we report the analytical performances of The Binding Site (TBS) SPA PLUS ® human IgGSc assay and the concordance with the Siemens BNII® human IgGSc assay. We evaluated precision, LoB, LoD and linearity of TBS SPA PLUS ® human IgGSc immunoassay. Quantitation of IgGSc in 53 patients' serum samples was performed in parallel on both analyzers. Results from both assays were compared. Analytical performances of the TBS SPA PLUS ® human IgGSc assay are acceptable for routine clinical use. According to the method comparison study, TBS assay measures lower values than Siemens assay for IgG1 and IgG4, whereas for IgG2 and IgG3 TBS provides greater values. All assays present a proportional bias, greater in the case of IgG3 and IgG4 assays. Individual subclass agreement, based on the classification of samples within three categories (low, normal and high) according to assay-specific reference intervals, range from 75% (IgG1) to 92% (IgG2). However, total classification agreement over all four subclasses only account for 55% of samples. Results obtained from both assays are not interchangeable. Standardization of IgGSc assay and review of the reference ranges must be accomplished in order to achieve a higher degree of agreement between different methods. Copyright © 2017 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  1. Challenges of Non-Destructive Assay Waste Measurement

    International Nuclear Information System (INIS)

    Shull, A.H.

    2003-01-01

    Historically, the Savannah River Site (SRS) routinely produced special nuclear material (SNM), which provided stable measurement conditions for the non-destructive assay (NDA) methods. However, the main mission of SRS has changed from the production of SNM to the processing of waste and material stabilization. Currently, the purpose of processing is to recover the SNM from the waste and stabilization materials, much of which is from other DOE facilities. These missions are usually of a short duration, but require non-destructive assay (NDA) accountability measurements on materials of varying composition and geometric configuration. These missions usually have cost and time constraints, which sometimes require re-application of existing NDA methods to waste measurements. Usually, each new material or re-application of the NDA method to a different SNM campaign requires new standards and timely re-calibration of the method. These constraints provide numerous challenges for the NDA methods, particularly in the area of measurement uncertainty. This paper will discuss the challenges of these situations, mainly from a measurement and statistical point of view and provide some possible solutions to the problems encountered. Specific examples will be discussed for the segmented gamma scanner (SGS), neutron multiplicity counter (NMC) and passive neutron coincidence counter (PNCC), which are some of the most common NDA instruments at SRS

  2. Application of neutron multiplicity counting to waste assay

    International Nuclear Information System (INIS)

    Pickrell, M.M.; Ensslin, N.

    1997-01-01

    This paper describes the use of a new figure of merit code that calculates both bias and precision for coincidence and multiplicity counting, and determines the optimum regions for each in waste assay applications. A tunable multiplicity approach is developed that uses a combination of coincidence and multiplicity counting to minimize the total assay error. An example is shown where multiplicity analysis is used to solve for mass, alpha, and multiplication and tunable multiplicity is shown to work well. The approach provides a method for selecting coincidence, multiplicity, or tunable multiplicity counting to give the best assay with the lowest total error over a broad spectrum of assay conditions

  3. Interdependence of initial cell density, drug concentration and exposure time revealed by real-time impedance spectroscopic cytotoxicity assay

    DEFF Research Database (Denmark)

    Caviglia, Claudia; Zor, Kinga; Canepa, Silvia

    2015-01-01

    We investigated the combined effect of the initial cell density (12 500, 35 000, 75 000, and 100 000 cells cm−2) and concentration of the anti-cancer drug doxorubicin on HeLa cells by performing timedependent cytotoxicity assays using real-time electrochemical impedance spectroscopy. A correlation...... between the rate of cell death and the initial cell seeding density was found at 2.5 μM doxorubicin concentration, whereas this was not observed at 5 or 100 μM. By sensing the changes in the cell–substrate interaction using impedance spectroscopy under static conditions, the onset of cytotoxicity...... was observed 5 h earlier than when using a standard colorimetric end-point assay (MTS) which measures changes in the mitochondrial metabolism. Furthermore, with the MTS assay no cytotoxicity was observed after 15 h of incubation with 2.5 μM doxorubicin, whereas the impedance showed at this time point cell...

  4. 125I anti-immunoglobulin binding assay for the detection and characterization of anti-platelet antibodies

    International Nuclear Information System (INIS)

    Kirkley, J.; Fabre, J.W.

    1980-01-01

    The binding assay as described in this paper is a very versatile system, and in this study it has been evaluated specifically for the detection of allo- or autoantibodies to platelets in man. The basic assay involves the incubation of a standard number of platelets with dilutions of test sera and the detection of platelet bound immunoglobulin by a second incubation with 125I labeled, immunoadsorbent purified rabbit F(ab') anti-human F(ab')2 (RAH). Of most importance, by varying the number of target platelets in the titrations and looking for binding plateaus, one can readily define conditions of optimum sensitivity for particular serum/platelet combinations. In addition, the assay can be used in conjunction with quantitative absorptions to subdivide complex sera into subspecificities and to give an estimate of the relative amounts of particular antigens on different platelets or other tissue or cell suspensions. One can also use saturating concentrations of RAH in the second incubation, in which case the amount of platelet bound radioactivity is directly related to the amount of first antibody bound to the platelets, and this can be manipulated to give information about serum antibody concentrations and amounts of antigen on the target tissue. The problem of ABO antibodies in this system, optimal conditions for platelet storage for the assay, and techniques for reducing assay backgrounds resulting from immunoglobulin adsorbed to the platelet surface are all evaluated

  5. Comparison of Microtox and Xenoassay Light as a Near Real Time River Monitoring Assay for Heavy Metals

    Directory of Open Access Journals (Sweden)

    M. I. E. Halmi

    2014-01-01

    Full Text Available Luminescence-based assays for toxicants such as Microtox, ToxAlert, and Biotox have been used extensively worldwide. However, the use of these assays in near real time conditions is limited due to nonoptimal assay temperature for the tropical climate. An isolate that exhibits a high luminescence activity in a broad range of temperatures was successfully isolated from the mackerel, Rastrelliger kanagurta. This isolate was tentatively identified as Photobacterium sp. strain MIE, based on partial 16S rDNA molecular phylogeny. Optimum conditions that support high bioluminescence activity occurred between 24 and 30°C, with pH 5.5 to 7.5, 10 to 20 g/L of sodium chloride, 30 to 50 g/L of tryptone, and 4 g/L of glycerol as the carbon source. Assessment of near real time capability of this bacterial system, Xenoassay light to monitor heavy metals from a contaminated river running through the Juru River Basin shows near real time capability with assaying time of less than 30 minutes per samples. Samples returned to the lab were tested with a standard Microtox assay using Vibrio fishceri. Similar results were obtained to Xenoassay light that show temporal variation of copper concentration. Thus, this strain is suitable for near real time river monitoring of toxicants especially in the tropics.

  6. Evaluation of the Elecsys® Chagas Assay for the Detection ofTrypanosoma cruzi-Specific Antibodies in a Multicenter Study in Europe and Latin America.

    Science.gov (United States)

    Flores-Chavez, Maria Delmans; Sambri, Vittorio; Schottstedt, Volkmar; Higuera-Escalante, Fernando Aparicio; Roessler, Dieter; Chaves, Monica; Laengin, Tina; Martinez, Alfredo; Fleischer, Bernhard

    2018-02-14

    Serology is the preferred method to confirm Chagas disease diagnosis and to screen blood donors. A battery of assays is often required due to the limited accuracy of single assays. The Elecsys® Chagas assay is a newly developed, double-antigen sandwich assay for use on the Elecsys® and cobas® e immunoassay analyzers, intended to identify individuals infected with Trypanosoma cruzi, for diagnosis and screening. The performance of the Elecsys® Chagas assay was evaluated in comparison with other widely used T. cruzi antibody assays, at multiple sites (Europe/Latin America). Relative sensitivity and specificity were assessed in samples from blood donors, pregnant women, and hospitalized patients, from endemic and non-endemic regions. The Elecsys® Chagas assay had an overall relative sensitivity of 100% ( n = 674). Overall relative specificity was 99.90% ( n = 14,681), 100% ( n = 313) and 100% ( n = 517) in samples from blood donors, pregnant women and hospitalized patients respectively. Analytical specificity was 99.83% ( n = 594). The Elecsys® Chagas assay detected T. cruzi antibodies in two World Health Organization (WHO)-standard T. cruzi reference panels (09/188 and 09/186) at a 1:512 dilution, corresponding to a cut-off sensitivity of approximately 1 mIU/ml. The Elecsys® Chagas assay demonstrated a robust performance under routine conditions at multiple sites in Europe and Latin America. In contrast to other available Chagas assays, the Elecsys® assay uses a reduced number of recombinant T. cruzi antigens resulting in a significantly smaller number of cross-reactions having improved analytical specificity, while being highly sensitive. Copyright © 2018 Flores-Chavez et al.

  7. Radioenzymatic assay of DOPA (3,4-dihydroxyphenylalanine)

    International Nuclear Information System (INIS)

    Johnson, G.A.; Gren, J.M.; Kupiecki, R.

    1978-01-01

    We modified the single-isotope radioenzymatic assay for catecholamines [Life Sci. 21, 625(1977)] to assay 3,4-dihydroxyphenylalanine (DOPA). DOPA decarboxylase is used to convert DOPA to dopamine, which concurrently is converted to [ 3 H]-3-O-methyldopamine in the presence of catechol-O-methyltransferase and [methyl- 3 H]-S-adenosylmethionine and assayed radioenzymatically. For assay of plasma DOPA, 50 μl of untreated plasma is added directly into the incubation mixture. A duplicate mixture containing an internal standard requires a second 50-μl aliquot of plasma. Because the assay measures both DOPA and endogenous dopamine, two additional aliquots of plasma must be assayed for dopamine in the absence of the decarboxylase by the differential assay; DOPA is estimated by difference. The assay is sensitive to 25 pg (500 ng/liter of plasma). Analysis of DOPA (DOPA plus dopamine) and the concurrent differential assay of catecholamines in at least 10 samples can be done in a single working day. Plasma DOPA concentrations for 42 normotensive adults were 1430 +- 19 ng/liter (mean +- SEM). In contrast, dopamine concentrations for these same subjects averaged 23 +- 20 ng/liter. Values for the 24 women subjects (1510 +- 62 ng/liter) significantly (P = 0.04) exceeded those for the men

  8. Intra-/inter-laboratory validation study on reactive oxygen species assay for chemical photosafety evaluation using two different solar simulators.

    Science.gov (United States)

    Onoue, Satomi; Hosoi, Kazuhiro; Toda, Tsuguto; Takagi, Hironori; Osaki, Naoto; Matsumoto, Yasuhiro; Kawakami, Satoru; Wakuri, Shinobu; Iwase, Yumiko; Yamamoto, Toshinobu; Nakamura, Kazuichi; Ohno, Yasuo; Kojima, Hajime

    2014-06-01

    A previous multi-center validation study demonstrated high transferability and reliability of reactive oxygen species (ROS) assay for photosafety evaluation. The present validation study was undertaken to verify further the applicability of different solar simulators and assay performance. In 7 participating laboratories, 2 standards and 42 coded chemicals, including 23 phototoxins and 19 non-phototoxic drugs/chemicals, were assessed by the ROS assay using two different solar simulators (Atlas Suntest CPS series, 3 labs; and Seric SXL-2500V2, 4 labs). Irradiation conditions could be optimized using quinine and sulisobenzone as positive and negative standards to offer consistent assay outcomes. In both solar simulators, the intra- and inter-day precisions (coefficient of variation; CV) for quinine were found to be below 10%. The inter-laboratory CV for quinine averaged 15.4% (Atlas Suntest CPS) and 13.2% (Seric SXL-2500V2) for singlet oxygen and 17.0% (Atlas Suntest CPS) and 7.1% (Seric SXL-2500V2) for superoxide, suggesting high inter-laboratory reproducibility even though different solar simulators were employed for the ROS assay. In the ROS assay on 42 coded chemicals, some chemicals (ca. 19-29%) were unevaluable because of limited solubility and spectral interference. Although several false positives appeared with positive predictivity of ca. 76-92% (Atlas Suntest CPS) and ca. 75-84% (Seric SXL-2500V2), there were no false negative predictions in both solar simulators. A multi-center validation study on the ROS assay demonstrated satisfactory transferability, accuracy, precision, and predictivity, as well as the availability of other solar simulators. Copyright © 2013 Elsevier Ltd. All rights reserved.

  9. Assessment and reduction of comet assay variation in relation to DNA damage: studies from the European Comet Assay Validation Group

    DEFF Research Database (Denmark)

    Møller, Peter; Möller, Lennart; Godschalk, Roger W L

    2010-01-01

    The alkaline single cell gel electrophoresis (comet) assay has become a widely used method for the detection of DNA damage and repair in cells and tissues. Still, it has been difficult to compare results from different investigators because of differences in assay conditions and because the data...... are reported in different units. The European Comet Assay Validation Group (ECVAG) was established for the purpose of validation of the comet assay with respect to measures of DNA damage formation and its repair. The results from this inter-laboratory validation trail showed a large variation in measured level...... reliability for the measurement of DNA damage by the comet assay but there is still a need for further validation to reduce both assay and inter-laboratory variation....

  10. Assay for methadone

    International Nuclear Information System (INIS)

    1980-01-01

    An improved radioimmunoassay for methadone is described using a novel antigen, antibody and labelled methadone derivatives. The preparation of a hemi-ester antigen is described by reacting a methadone derivative with succinic anhydride or glutaric anhydride; this hapten is then covalently bonded through the carboxyl group to bovine serum albumin. An antibody specific to methadone is produced by inoculating a host animal with the above antigen. The unknown amount of methadone in a sample is then determined by mixing the sample with a known amount of radiolabelled methadone derivative and the above antibody and comparing the degree of binding to a standard curve obtained by mixing the antibody with known amounts of methadone and fixed amounts of labelled methadone derivative. The radioimmunoassay was used to measure methadone levels in urine from individuals attending a methadone clinic. (U.K.)

  11. Study on quantification of HBs-antibody by immunoradiometric assay

    International Nuclear Information System (INIS)

    Kondo, Yuichi; Itoi, Yoshihiro; Kajiyama, Shizuo

    1989-01-01

    Quantification of HBs-antibody assay was carried out using a commercialized assay kit and standard solutions of HBs-antibody recognised as 1 st reference preparation of hepatitis B immunogloblin by WHO. Standard curve of HBs-antibody was drawn with the function of 3D-spline and the correlation factor was obtained as r = 0.999. Coefficient of intra-assay variance was 3.8 % and that of inter-assay variance was 7.8 %. Dilution tests showed satisfactory results in the range of 2-16 times. Correlation between value of cut-off indices and concentration of HBs-antibody was obtained as the formula of y = 2.599 x-3.894 (r = 0.992) and 2.1 of cut-off index corresponded to about 5 mIU/ml of HBs-antibody concentration. (author)

  12. Phosphorimaging detection and quantitation for isotopic ion flux assays.

    Science.gov (United States)

    Fitch, Richard W; Daly, John W

    2005-07-15

    A 96-well-microplate-based ion flux method utilizing readily available autoradiographic phosphorimaging detection is described. Nicotinic acetylcholine receptor-mediated (22)Na influx in four cultured cell lines provided satisfactory concentration-response data for epibatidine and several other nicotinic agonists. The data were consistent with data obtained using standard 6-well assays. Assays for nicotinic-receptor-mediated (86)Rb efflux produced data similar to data obtained with the (22)Na influx assay. However, assays for (45)Ca influx were not successful, although (45)Ca was readily detected and quantified. Voltage-gated sodium channel-mediated (22)Na influx in a neuroblastoma cell line allowed assay of the effects of such sodium channel activators as batrachotoxin and a pumiliotoxin B/scorpion venom combination. Phosphorimaging detection allows for reliable beta counting of up to 1,200 simultaneous samples with excellent sensitivity and is amenable for application to high-throughput screening.

  13. The development of SRM assays is transforming proteomics research.

    Science.gov (United States)

    Manes, Nathan P; Nita-Lazar, Aleksandra

    2017-04-01

    Bottom-up targeted proteomics using SRM is a powerful analytical technology, but it requires the development of SRM assays, which is a complex procedure. Whereas proteome-wide SRM assays have recently been developed for a small number of species, this is not so for the mouse. In this issue, Percy et al. report the development of hundreds of mouse SRM assays. Their development required shotgun MS to identify proteotypic peptides, synthesis, and LC-MS characterization of peptide standards, and interlaboratory SRM to robustly assess the quality of the assays. The resulting SRM assays are intended to be used to analyze mouse plasma and cardiac tissue, primarily for cardiovascular disease and cancer research. Published 2016. This article is a U.S. Government work and is in the public domain in the USA.

  14. MS transport assays for γ-aminobutyric acid transporters--an efficient alternative for radiometric assays.

    Science.gov (United States)

    Schmitt, Sebastian; Höfner, Georg; Wanner, Klaus T

    2014-08-05

    Transport assays for neurotransmitters based on radiolabeled substrates are widely spread and often indispensable in basic research and the drug development process, although the use of radioisotopes is inherently coupled to issues concerning radioactive waste and safety precautions. To overcome these disadvantages, we developed mass spectrometry (MS)-based transport assays for γ-aminobutyric acid (GABA), which is the major inhibitory neurotransmitter in the central nervous system (CNS). These "MS Transport Assays" provide all capabilities of [(3)H]GABA transport assays and therefore represent the first substitute for the latter. The performance of our approach is demonstrated for GAT1, the most important GABA transporter (GAT) subtype. As GABA is endogenously present in COS-7 cells employed as hGAT1 expression system, ((2)H6)GABA was used as a substrate to differentiate transported from endogenous GABA. To record transported ((2)H6)GABA, a highly sensitive, short, robust, and reliable HILIC-ESI-MS/MS quantification method using ((2)H2)GABA as an internal standard was developed and validated according to the Center for Drug Evaluation and Research (CDER) guidelines. Based on this LC-MS quantification, a setup to characterize hGAT1 mediated ((2)H6)GABA transport in a 96-well format was established, that enables automated processing and avoids any sample preparation. The K(m) value for ((2)H6)GABA determined for hGAT1 is in excellent agreement with results obtained from [(3)H]GABA uptake assays. In addition, the established assay format enables efficient determination of the inhibitory potency of GAT1 inhibitors, is capable of identifying those inhibitors transported as substrates, and furthermore allows characterization of efflux. The approach described here combines the strengths of LC-MS/MS with the high efficiency of transport assays based on radiolabeled substrates and is applicable to all GABA transporter subtypes.

  15. Simultaneous assay of every Salmonella Typhi gene using one million transposon mutants

    Science.gov (United States)

    Langridge, Gemma C.; Phan, Minh-Duy; Turner, Daniel J.; Perkins, Timothy T.; Parts, Leopold; Haase, Jana; Charles, Ian; Maskell, Duncan J.; Peters, Sarah E.; Dougan, Gordon; Wain, John; Parkhill, Julian; Turner, A. Keith

    2009-01-01

    Very high-throughput sequencing technologies need to be matched by high-throughput functional studies if we are to make full use of the current explosion in genome sequences. We have generated a very large bacterial mutant pool, consisting of an estimated 1.1 million transposon mutants and we have used genomic DNA from this mutant pool, and Illumina nucleotide sequencing to prime from the transposon and sequence into the adjacent target DNA. With this method, which we have called TraDIS (transposon directed insertion-site sequencing), we have been able to map 370,000 unique transposon insertion sites to the Salmonella enterica serovar Typhi chromosome. The unprecedented density and resolution of mapped insertion sites, an average of one every 13 base pairs, has allowed us to assay simultaneously every gene in the genome for essentiality and generate a genome-wide list of candidate essential genes. In addition, the semiquantitative nature of the assay allowed us to identify genes that are advantageous and those that are disadvantageous for growth under standard laboratory conditions. Comparison of the mutant pool following growth in the presence or absence of ox bile enabled every gene to be assayed for its contribution toward bile tolerance, a trait required of any enteric bacterium and for carriage of S. Typhi in the gall bladder. This screen validated our hypothesis that we can simultaneously assay every gene in the genome to identify niche-specific essential genes. PMID:19826075

  16. Dendritic cell migration assay: a potential prediction model for identification of contact allergens.

    Science.gov (United States)

    Gibbs, Susan; Spiekstra, Sander; Corsini, Emanuela; McLeod, Julie; Reinders, Judith

    2013-04-01

    This manuscript describes methodology and a prediction model for the MUTZ-LC migration assay. The assay represents the physiological change in Langerhans cell (LC) behavior after exposure to a sensitizing chemical, resulting in LC migration from the epidermis to the dermis. MUTZ-LC are derived from the commercially available MUTZ-3 cell line. Upon exposure to a sensitizer MUTZ-LC migrate preferentially towards CXCL12 whereas upon exposure to a non-sensitizer MUTZ-LC migrate towards CCL5. A CXCL12/CCL5 ratio >1.10 in 2/3 independent experiments is indicative of a sensitizer, whereas a CXCL12/CCL5 ratio ≤1.10 is indicative of a non-sensitizer. At non cytotoxic chemical concentrations 9 sensitizers (2,4-dinitrochlorobenzene, paraphenylendiamine, cinnamaldehyde, isoeugenol, nickel-sulfate, tetramethylthiuram disulfide, eugenol, cinnamic-alcohol, ammonium-hexachloroplatinate) were distinguished from 4 non sensitizers (sodium lauryl sulfate, salicylic acid, phenol, octanoic acid). Critical points in assay performance are (i) MUTZ-3 passage number after thawing (p6-p40); (ii) cell viability (>80%); (iii) standard curve to optimize correlation of fluorescence with cell number; and (iv) optimization of the concentration of rhCXCL12 and rhCCL5 in transwell. The protocol has been tested in three European laboratories and results suggest that it may provide working conditions for performing the DC migration assay which is aimed at distinguishing sensitizers from non sensitizers. Copyright © 2012 Elsevier Ltd. All rights reserved.

  17. Monkey Feeding Assay for Testing Emetic Activity of Staphylococcal Enterotoxin.

    Science.gov (United States)

    Seo, Keun Seok

    2016-01-01

    Staphylococcal enterotoxins (SEs) are unique bacterial toxins that cause gastrointestinal toxicity as well as superantigenic activity. Since systemic administration of SEs induces superantigenic activity leading to toxic shock syndrome that may mimic enterotoxic activity of SEs such as vomiting and diarrhea, oral administration of SEs in the monkey feeding assay is considered as a standard method to evaluate emetic activity of SEs. This chapter summarizes and discusses practical considerations of the monkey feeding assay used in studies characterizing classical and newly identified SEs.

  18. 18 CFR 157.206 - Standard conditions.

    Science.gov (United States)

    2010-04-01

    ... Discharge Elimination System Program, 40 CFR part 122 et seq.; (ii) Clean Air Act, as amended (42 U.S.C... this section only if it adheres to Commission staff's current “Upland Erosion Control, Revegetation and... any new compressor station, compression added to an existing station, or any modification, upgrade or...

  19. Bioelectrocatalyzed Nitrogen Fixation under Standard Conditions

    Science.gov (United States)

    2016-11-07

    FEMS Microbiology Letters . 1984, 10, 299- 302. 3. Kumar, Ashok; Tabita, Robert F.; Van Baalen, Chase. High endogenous nitrogenase activity in...York, New York, 1985, pp 129-138. 12. Tsygankov, A. A. Nitrogen-Fixing Cyanobacteria: A Review. Applied Biochemistry and Microbiology . 2007, 43, 250...scholarships or fellowships for further studies in science, mathematics, engineering or technology fields: Student Metrics This section only applies to

  20. A simple and fast kinetic assay for the determination of fructan exohydrolase activity in perennial ryegrass (Lolium perenne L.

    Directory of Open Access Journals (Sweden)

    Anna eGasperl

    2015-12-01

    Full Text Available Despite the fact that fructans are the main constituent of water-soluble carbohydrates in forage grasses and cereal crops of temperate climates, little knowledge is available on the regulation of the enzymes involved in fructan metabolism. The analysis of enzyme activities involved in this process has been hampered by the low affinity of the fructan enzymes for sucrose and fructans used as fructosyl donor. Further, the analysis of fructan composition and enzyme activities is restricted to specialized labs with access to suited HPLC equipment and appropriate fructan standards. The degradation of fructan polymers with high degree of polymerization (DP by fructan exohydrolases (FEHs to fructosyloligomers is important to liberate energy in the form of fructan, but also under conditions where the generation of low DP polymers is required. Based on published protocols employing enzyme coupled endpoint reactions in single cuvettes, we developed a simple and fast kinetic 1-FEH assay. This assay can be performed in multi-well plate format using plate readers to determine the activity of 1-FEH against 1-kestotriose, resulting in a significant time reduction. Kinetic assays allow an optimal and more precise determination of enzyme activities compared to endpoint assays, and enable to check the quality of any reaction with respect to linearity of the assay. The enzyme coupled kinetic 1-FEH assay was validated in a case study showing the expected increase in 1-FEH activity during cold treatment. This assay is cost effective and could be performed by any lab with access to a plate reader suited for kinetic measurements and readings at 340 nm, and is highly suited to assess temporal changes and relative differences in 1-FEH activities. Thus, this enzyme coupled kinetic 1-FEH assay is of high importance both to the field of basic fructan research and plant breeding.

  1. An IMS/ATP Assay for the Detection of Mycobacterium tuberculosis in Urine

    Directory of Open Access Journals (Sweden)

    Dawn M. Hunter

    2012-01-01

    Full Text Available Background. Although sputum smears are the gold standard for diagnosis of tuberculosis, sensitivity in HIV/TB coinfection cases is low, indicating a need for alternative methods. Urine is being increasingly evaluated. Materials and Methods. A novel method for detecting Mycobacterium tuberculosis (MTB in synthetic urine using a combined IMS/ATP assay was evaluated. Preliminary work established standard ATP conditions and the sensitivity and specificity of the MTB antibody. Eighty-four blinded samples in four replicate assays were evaluated for the presence of MTB using labeled immunomagnetic beads for capture. Beads were separated, washed, and resuspended in broth and added to a microtiter plate. Bioluminescent output was measured and signal-to-noise ratios were calculated. All samples were plated on Middlebrook 7H10 agar or trypticase soy agar to determine limit of detection and recoveries. Results and Conclusions. MTB was distinguished from common bacteriuria isolates and other nontarget bacteria by its ATP results. IMS/ATP successfully detected 19 of 28 samples of MTB in synthetic urine with a limit of detection of 104 CFU/ml. Sensitivity and specificity were 67.9% and 82.1%, respectively. This assay offers a possible rapid screening method for HIV-positive patients with suspected coinfection to improve MTB diagnosis.

  2. Nondestructive analysis of nuclear materials by isotopic source assay system

    International Nuclear Information System (INIS)

    Masui, Jinichi; Tuboya, Takao

    1978-01-01

    Destructive assay is an effective method for the analysis of nuclear materials in nuclear fuels, but is not suitable for some aspects of nuclear fuel cycle, for example, for accounting and control or safeguard purposes. Isotopic Source Assay System was imported from Intelcom Rad. Tech. Company, and the results of assay of enriched uranium and plutonium sealed for passive and active assay by the system are presented. A 252 Cf source is provided for the assay system. Assay of unknown samples by this system is carried out relatively to the measurement of known standards. Several known standards approximating the physical and chemical properties of unknown samples are prepared to make calibration curves. When one fission event occurs in a sample, a few neutrons (2.5 neutrons on the average) and gamma ray (about 7 photons) are emitted simultaneously. By three detector coincidence out of four, one count is registered by the assay system. First, statistical informations and geometry were examined. Then, three kinds of enriched uranium were measured to examine the measurement on 238 U. Passive and active measurements were performed on 4.32 grams of PuO 2 during one month to know reproducibility. In conclusion of these tests, it was proved to be able to apply this system to the analysis of nuclear materials similar in enrichment or isotopic composition, and scraps and wastes containing known matrix materials. (Wakatsuki, Y.)

  3. A Morphological identification cell cytotoxicity assay using cytoplasm-localized fluorescent probe (CLFP) to distinguish living and dead cells.

    Science.gov (United States)

    Lai, Fangfang; Shen, Zhengwei; Wen, Hui; Chen, Jialing; Zhang, Xiang; Lin, Ping; Yin, Dali; Cui, Huaqing; Chen, Xiaoguang

    2017-01-08

    Cell cytotoxicity assays include cell activity assays and morphological identification assays. Currently, all frequently used cytotoxicity assays belong to cell activity assays but suffer from detection limitations. Morphological identification of cell death remains as the gold standard, although the method is difficult to scale up. At present there is no generally accepted morphological identification based cell cytotoxicity assay. In this study, we applied previous developed cell cytoplasm-localized fluorescent probe (CLFP) to display cell morphologies. Under fluorescence microscopy, the fluorescence morphology and intensity of living cells are distinct from dead cells. Based on these characters we extracted the images of living cells from series of samples via computational analysis. Thus, a novel cell morphological identification cytotoxicity assay (CLFP assay) is developed. The performance of the CLFP assay was similar to cell activity assay (MTT assay), but the accuracy of the CLFP assay was superior when measuring the cytotoxicity of active compounds. Copyright © 2016 Elsevier Inc. All rights reserved.

  4. ARIES nondestructive assay system operation and performance

    International Nuclear Information System (INIS)

    Cremers, Teresa L.; Hansen, Walter J.; Herrera, Gary D.; Nelson, David C.; Sampson, Thomas E.; Scheer, Nancy L.

    2000-01-01

    The ARIES (Advanced Recovery and Integrated Extraction System) Project is an integrated system at the Los Alamos Plutonium Facility for the dismantlement of nuclear weapons. The plutonium produced by the ARIES process was measured by an integrated nondestructive assay (NDA) system. The performance of the NDA systems was monitored by a measurement control program which is a part of a nuclear material control and accountability system. In this paper we will report the results of the measurements of the measurement control standards as well as an overview of the measurement of the ARIES process materials

  5. Automatic titrator for high precision plutonium assay

    International Nuclear Information System (INIS)

    Jackson, D.D.; Hollen, R.M.

    1986-01-01

    Highly precise assay of plutonium metal is required for accountability measurements. We have developed an automatic titrator for this determination which eliminates analyst bias and requires much less analyst time. The analyst is only required to enter sample data and start the titration. The automated instrument titrates the sample, locates the end point, and outputs the results as a paper tape printout. Precision of the titration is less than 0.03% relative standard deviation for a single determination at the 250-mg plutonium level. The titration time is less than 5 min

  6. Extraction-free cortisol assay

    International Nuclear Information System (INIS)

    1980-01-01

    A method for determining the concentration of cortisol in a serum sample comprises: (a) incubating the sample, labelled cortisol and an effective amount of deblocking agent in an aqueous medium with a composite comprising anti-cortisol antibodies fixed in active form onto the surfaces of a negatively charged support material, the incubation being carried out at a pH of from 4.0 to 6.5 and under conditions sufficient to result in the formation of immunochemical complexes on the composite, some of which complexes comprise labelled cortisol; (b) separating the composite from the incubation medium; (c) determining the amount of label on the separated composite or in the remaining incubation medium; and (d) relating the determination of (c) to a standard to determine the cortisol concentration in the sample. (author)

  7. Hyperpolarized NMR Probes for Biological Assays

    Directory of Open Access Journals (Sweden)

    Sebastian Meier

    2014-01-01

    Full Text Available During the last decade, the development of nuclear spin polarization enhanced (hyperpolarized molecular probes has opened up new opportunities for studying the inner workings of living cells in real time. The hyperpolarized probes are produced ex situ, introduced into biological systems and detected with high sensitivity and contrast against background signals using high resolution NMR spectroscopy. A variety of natural, derivatized and designed hyperpolarized probes has emerged for diverse biological studies including assays of intracellular reaction progression, pathway kinetics, probe uptake and export, pH, redox state, reactive oxygen species, ion concentrations, drug efficacy or oncogenic signaling. These probes are readily used directly under natural conditions in biofluids and are often directly developed and optimized for cellular assays, thus leaving little doubt about their specificity and utility under biologically relevant conditions. Hyperpolarized molecular probes for biological NMR spectroscopy enable the unbiased detection of complex processes by virtue of the high spectral resolution, structural specificity and quantifiability of NMR signals. Here, we provide a survey of strategies used for the selection, design and use of hyperpolarized NMR probes in biological assays, and describe current limitations and developments.

  8. ['Gold standard', not 'golden standard'

    NARCIS (Netherlands)

    Claassen, J.A.H.R.

    2005-01-01

    In medical literature, both 'gold standard' and 'golden standard' are employed to describe a reference test used for comparison with a novel method. The term 'gold standard' in its current sense in medical research was coined by Rudd in 1979, in reference to the monetary gold standard. In the same

  9. Developments in plutonium waste assay at AWE

    International Nuclear Information System (INIS)

    Miller, T J

    2009-01-01

    In 2002 a paper was presented at the 43rd Annual Meeting of the Institute of Nuclear Materials Management (INMM) on the assay of low level plutonium (Pu) in soft drummed waste (Miller 2002 INMM Ann. Meeting (Orlando, FL, 23-27 July 2002)). The technique described enabled the Atomic Weapons Establishment (AWE), at Aldermaston in the UK, to meet the stringent Low Level Waste Repository at Drigg (LLWRD) conditions for acceptance for the first time. However, it was initially applied to only low density waste streams because it relied on measuring the relatively low energy (60 keV) photon yield from Am-241 during growth. This paper reviews the results achieved when using the technique to assay over 10 000 waste packages and presents the case for extending the range of application to denser waste streams.

  10. A Quantitative Fluorescence-Based Lipase Assay

    Directory of Open Access Journals (Sweden)

    Giovanna Lomolino

    2012-01-01

    Full Text Available An easy and fast gel diffusion assay for detecting and monitoring lipase activity by quantification of fluorescein is described. By measuring the intensity of fluorescein, it is possible to obtain a calibration curve with a regression coefficient better than by using the radius of fluorescent haloes. Through the quantification of fluorescence intensity of fluorescein released after the hydrolysis of a fluorescent ester, fluorescein dibutyrate, used as substrate in agar plates, commercial and skimmed milk lipase activity were studied. Moreover, with this method, lipase activity can be monitored in reaction medium that contains compounds which are affected by turbidity or cause measurement interference for UV-spectrophotometer and fluorimeter. In this experiment, boiled skimmed milk was dispersed in the agar gel with fluorescein dibutyrate, and it was used as a reaction medium to mimic natural conditions. The development of such an assay has a potential for applications in industries ranging from pharmaceuticals to food production and monitoring.

  11. Standardization of dot-ELISA for the serological diagnosis of toxocariasis and comparision of the assay with ELISA Padronização do teste dot-ELISA para o diagnóstico da toxocaríase, estudo comparativo com o teste imunoenzimático ELISA

    Directory of Open Access Journals (Sweden)

    Eide Dias Camargo

    1992-02-01

    Full Text Available The dot-enzyme-linked immunosorbent assay (dot-ELISA was standardized using somatic (S and excretory-secretory (ES antigens of Toxocara-canis for the detection of specific antibodies in 22 serum samples from children aged 1 to 15 years, with clinical signs of toxocariasis. Fourteen serum samples from apparently normal individuals and 28 sera from patients with other pathologies were used as controls. All samples were used before and after absorption with Ascaris suum extract. When the results were evaluated in comparison with ELISA, the two tests were found to have similar sensitivity, but dot-ELISA was found to be more specific in the presence of the two antigens studied. Dot-ELISA proved to be effective for the diagnosis of human toxocariasis, presenting advantages in terms of yield, stability, time and ease of execution and low cost.Padronizou-se o teste imunoenzimático dot-ELISA, empregando-se os antígenos somático (S e excretor-secretor (ES de Toxocara canis, para pesquisa de anticorpos específicos em 22 soros de pacientes com idades entre 5 a 15 anos, com dados clínicos de toxocaríase. Como grupo controle, foram estudados 14 soros de indivíduos supostamente normais e 28 soros de pacientes com outras patologias. Todas as amostras em estudo foram empregadas antes e após absorção com extrato de Ascaris suum. Os resultados obtidos foram avaliados comparativamente com o teste ELISA evidenciando, nos dois testes estudados, comportamento semelhante quanto à sensibilidade e maior especificidade para o dot-ELISA, com qualquer dos dois antígenos estudados. O teste dot-ELISA mostrou-se eficiente para o diagnóstico da toxocaríase humana, apresentando vantagens quanto ao rendimento, estabilidade, tempo e facilidade de execução e baixo custo.

  12. The fluorometric microculture cytotoxicity assay.

    Science.gov (United States)

    Lindhagen, Elin; Nygren, Peter; Larsson, Rolf

    2008-01-01

    The fluorometric microculture cytotoxicity assay (FMCA) is a nonclonogenic microplate-based cell viability assay used for measurement of the cytotoxic and/or cytostatic effect of different compounds in vitro. The assay is based on hydrolysis of the probe, fluorescein diacetate (FDA) by esterases in cells with intact plasma membranes. The assay is available as both a semiautomated 96-well plate setup and a 384-well plate version fully adaptable to robotics. Experimental plates are prepared with a small amount of drug solution and can be stored frozen. Cells are seeded on the plates and cell viability is evaluated after 72 h. The protocol described here is applicable both for cell lines and freshly prepared tumor cells from patients and is suitable both for screening in drug development and as a basis for a predictive test for individualization of anticancer drug therapy.

  13. Radioligand assay in reproductive biology

    International Nuclear Information System (INIS)

    Korenman, S.G.; Sherman, B.M.

    1975-01-01

    Radioligand assays have been developed for the principal reproductive steroids and peptide hormones. Specific binding reagents have included antibodies, plasma binders, and intracellular receptors. In each assay, problems of specificity, sensitivity, and nonspecific inhibitors were encountered. Many features of the endocrine physiology in childhood, during puberty, and in adulthood have been characterized. Hormonal evaluations of endocrine disorders of reproduction are characterized on the basis of their characteristic pathophysiologic alterations. (U.S.)

  14. Accounting standards

    NARCIS (Netherlands)

    Stellinga, B.; Mügge, D.

    2014-01-01

    The European and global regulation of accounting standards have witnessed remarkable changes over the past twenty years. In the early 1990s, EU accounting practices were fragmented along national lines and US accounting standards were the de facto global standards. Since 2005, all EU listed

  15. Digital microfluidic assay for protein detection.

    Science.gov (United States)

    Mok, Janine; Mindrinos, Michael N; Davis, Ronald W; Javanmard, Mehdi

    2014-02-11

    Global studies of the human proteome have revealed a plethora of putative protein biomarkers. However, their application for early disease detection remains at a standstill without suitable methods to realize their utility in the clinical setting. There thus continues to be tremendous interest in developing new technology for sensitive protein detection that is both low in cost and carries a small footprint to be able to be used at the point of care. The current gold standard method for protein biomarker detection is the ELISA, which measures protein abundance using bulky fluorescent scanners that lack portability. Here, we present a digital microfluidic platform for protein biomarker detection that is low in cost compared with standard optical detection methods, without any compromise in sensitivity. This platform furthermore makes use of simple electronics, enabling its translation into a portable handheld device, and has been developed in a manner that can easily be adapted to assay different types of proteomic biomarkers. We demonstrate its utility in quantifying not only protein abundance, but also activity. Interleukin-6 abundance could be assayed from concentrations as low as 50 pM (an order of magnitude lower than that detectable by a comparable laboratory designed ELISA) using less than 5 μL of sample, and Abelson tyrosine kinase activity was detectable in samples containing 100 pM of kinase.

  16. Correlation between the genotoxicity endpoints measured by two different genotoxicity assays: comet assay and CBMN assay

    Directory of Open Access Journals (Sweden)

    Carina Ladeira

    2015-06-01

    The results concerning of positive findings by micronuclei and non significant ones by comet assay, are corroborated by Deng et al. (2005 study performed in workers occupationally exposed to methotrexate, also a cytostatic drug. According to Cavallo et al. (2009, the comet assay seems to be more suitable for the prompt evaluation of the genotoxic effects, for instance, of polycyclic aromatic hydrocarbons mixtures containing volatile substances, whereas the micronucleus test seems more appropriate to evaluate the effects of exposure to antineoplastic agents. However, there are studies that observed an increase in both the comet assay and the micronucleus test in nurses handling antineoplastic drugs, although statistical significance was only seen in the comet assay, quite the opposite of our results (Maluf & Erdtmann, 2000; Laffon et al. 2005.

  17. Complementing in vitro screening assays with in silico ...

    Science.gov (United States)

    High-throughput in vitro assays offer a rapid, cost-efficient means to screen thousands of chemicals across hundreds of pathway-based toxicity endpoints. However, one main concern involved with the use of in vitro assays is the erroneous omission of chemicals that are inactive under assay conditions but that can generate active metabolites under in vivo conditions. To address this potential issue, a case study will be presented to demonstrate the use of in silico tools to identify inactive parents with the ability to generate active metabolites. This case study used the results from an orthogonal assay designed to improve confidence in the identification of active chemicals tested across eighteen estrogen receptor (ER)-related in vitro assays by accounting for technological limitations inherent within each individual assay. From the 1,812 chemicals tested within the orthogonal assay, 1,398 were considered inactive. These inactive chemicals were analyzed using Chemaxon Metabolizer software to predict the first and second generation metabolites. From the nearly 1,400 inactive chemicals, over 2,200 first-generation (i.e., primary) metabolites and over 5,500 second-generation (i.e., secondary) metabolites were predicted. Nearly 70% of primary metabolites were immediately detoxified or converted to other metabolites, while over 70% of secondary metabolites remained stable. Among these predicted metabolites, those that are most likely to be produced and remain

  18. Diagnosis of Morquio Syndrome in Dried Blood Spots Based on a New MRM-MS Assay.

    Directory of Open Access Journals (Sweden)

    Claudia Cozma

    Full Text Available Mucopolysaccharidosis IVA (MPS IVA; Morquio A disease is an autosomal recessive disease caused and characterized by a decreased activity of N-acetylgalactosamine-6-sulfate sulfatase (GALNS, resulting in accumulation of keratan sulfate and chondroitin-6-sulfate in tissues and secondary organ damage. Recently approved enzyme replacement therapy renders the easy and early identification of MPS IVA of out-most importance.We propose a completely new assay for the stable and reproducible detection of GALNS deficiency in dry blood spots (DBS. For the validation blood samples were taken from 59 healthy individuals and 24 randomly selected genetically confirmed MPS IVA patients. The material extracted from DBS was incubated with a 4-methylumbelliferyl-β-D-galactopyranoside-6-sulfate as a specific substrate. Final enzymatic product, 4-methylumbelliferone, obtained after adding exogenous beta-galactosidase, was quantified by LC/MRM-MS (liquid-chromatography/multiple-reaction-monitoring mass-spectrometry. 4-propyl-5-hydroxy-7-methyl-2h-chromen-2-one was used as internal standard, a compound with a similar molecular structure and fragmentation pattern in negative ion mode as 4-methylumbelliferone.The enzymatic assay yielded a positive and negative predictive value of 1.0 for genetically confirmed MPS IVA patients (GALNS activity of 0.35 ± 0.21 μmol/L/h and for controls with normal GALNS activity (23.1 ± 5.3 μmol/L /h. With present enzymatic conditions, the reaction yield in dried blood spots is at least 20 fold higher than any previously reported data with other assays.The present LC/MRM-MS based assay for MPS IVA diagnosis provides an easy, highly-standardized, accurate and innovative quantification of the enzymatic product in vitro and distinguishes perfectly between MPS IVA affected patients and normal controls. This technique will significantly simplify the early detection of MPS IVA patients.

  19. Diagnosis of Morquio Syndrome in Dried Blood Spots Based on a New MRM-MS Assay.

    Science.gov (United States)

    Cozma, Claudia; Eichler, Sabrina; Wittmann, Gyula; Flores Bonet, Alba; Kramp, Guido Johannes; Giese, Anne-Katrin; Rolfs, Arndt

    2015-01-01

    Mucopolysaccharidosis IVA (MPS IVA; Morquio A disease) is an autosomal recessive disease caused and characterized by a decreased activity of N-acetylgalactosamine-6-sulfate sulfatase (GALNS), resulting in accumulation of keratan sulfate and chondroitin-6-sulfate in tissues and secondary organ damage. Recently approved enzyme replacement therapy renders the easy and early identification of MPS IVA of out-most importance. We propose a completely new assay for the stable and reproducible detection of GALNS deficiency in dry blood spots (DBS). For the validation blood samples were taken from 59 healthy individuals and 24 randomly selected genetically confirmed MPS IVA patients. The material extracted from DBS was incubated with a 4-methylumbelliferyl-β-D-galactopyranoside-6-sulfate as a specific substrate. Final enzymatic product, 4-methylumbelliferone, obtained after adding exogenous beta-galactosidase, was quantified by LC/MRM-MS (liquid-chromatography/multiple-reaction-monitoring mass-spectrometry). 4-propyl-5-hydroxy-7-methyl-2h-chromen-2-one was used as internal standard, a compound with a similar molecular structure and fragmentation pattern in negative ion mode as 4-methylumbelliferone. The enzymatic assay yielded a positive and negative predictive value of 1.0 for genetically confirmed MPS IVA patients (GALNS activity of 0.35 ± 0.21 μmol/L/h) and for controls with normal GALNS activity (23.1 ± 5.3 μmol/L /h). With present enzymatic conditions, the reaction yield in dried blood spots is at least 20 fold higher than any previously reported data with other assays. The present LC/MRM-MS based assay for MPS IVA diagnosis provides an easy, highly-standardized, accurate and innovative quantification of the enzymatic product in vitro and distinguishes perfectly between MPS IVA affected patients and normal controls. This technique will significantly simplify the early detection of MPS IVA patients.

  20. Extracellular Enzyme Activity assay as indicator of soil microbial functional diversity and activity

    DEFF Research Database (Denmark)

    Hendriksen, Niels Bohse; Winding, Anne

    2012-01-01

    and soil ecosystem services. The soil enzyme activity has been measured by the use of fluorogenic model substrates e.g. methylumbelliferyl (MUF) substrates for a number of enzymes involved in the degradation of polysaccharides as cellulose, hemicellulose and chitin, while degradation of proteins has been......Extracellular Enzyme Activity assay as indicator of soil microbial functional diversity and activity Niels Bohse Hendriksen, Anne Winding. Department of Environmental Science, Aarhus University, 4000 Roskilde, Denmark Soil enzymes originate from a variety of organisms, notably fungi and bacteria......, experimental conditions of extraction of enzymes from soils, buffer and pH, substrate concentration, temperature and the necessary controls were optimized and standardized. This has resulted in an optimized standard operating procedure of EEA, which are being tested as an indicator of soil functional diversity...

  1. Radionuclide assay of membrane Na+, K+-ATPase activity of peserved red blood cells

    International Nuclear Information System (INIS)

    Trusov, V.V.; Zelenin, A.A.; Marizin, S.A.

    1986-01-01

    The radionuclide tests were used to investigate the influence of varying blood preservatives on erythrocylic membrane Na + , K + -ATPase activity in samples of whole blood and packed red blood cells from normal donors prepared by standard methods. The tests were performed before and after seven days of preservation under standard conditions. It was found that blood preservations lowered membrane Na + , K + -ATPase activity: its minimum reduction was recorded with citroglucopnosphate, while glugicir induced a significant drop in Na + , K + -ATPase activity of preserved red blood cells regardless of the type of the blood transfusion solution. The assay of membrane Na + , K + -ATPase activity of preserved red blood cells with the use of 86 Rb could be recommended as an evaluation test for preserved blood and its components

  2. Phagocytosis and killing assays for Candida species.

    Science.gov (United States)

    Du, Chen; Calderone, Richard A

    2009-01-01

    Both innate resistance and acquired cell-mediated immunity are involved in an anti-Candida response. Essential components of both the arms of the immune defense against infections by Candida spp. include phagocytic cells, i.e., polymorphonuclear neutrophils (PMNs) and mononuclear phagocytes. A powerful in vitro assay to assess host-pathogen interactions and study pathogenesis is the co-culture of phagocytic cells with a test fungus. The precise contribution of phagocytes to the host defense is usually assessed by determining phagocytosis and killing of Candida spp. blastoconidia. Dissection of the roles of various virulence factors in the infection process will involve the use of both in vitro and ex vivo assays. These assays are very useful as one of the approaches to determine the virulence factors of Candida spp., now that specific gene mutants are relatively easy to construct. In vitro studies involving specific cultured immune system cells can permit the analysis of interactions under controlled conditions. These studies provide an opportunity to monitor and compare host cell behavior upon challenge with wild-type or mutant strains of the pathogen.

  3. Validation of a KHV antibody enzyme-linked immunosorbent assay (ELISA)

    DEFF Research Database (Denmark)

    Bergmann, S M; Wang, Q; Zeng, W

    2017-01-01

    fully validated for KHV. Therefore, it was developed and validated an enzyme-linked immunosorbent assay (ELISA) to detect KHV antibodies. The assay was optimized with respect to plates, buffers, antigens and assay conditions. It demonstrated high diagnostic and analytical sensitivity and specificity...... and was particularly useful at the pond or farm levels. Considering the scale of the carp and koi industry worldwide, this assay represents an important practical tool for the indirect detection of KHV, also in the absence of clinical signs....

  4. Transfer of a two-tiered keratinocyte assay: IL-18 production by NCTC2544 to determine the skin sensitizing capacity and epidermal equivalent assay to determine sensitizer potency

    DEFF Research Database (Denmark)

    Teunis, Marc; Corsini, Emanuela; Smits, Mieke

    2013-01-01

    in a non-coded fashion. Here we describe the transferability to naïve laboratories, the establishment of the standard operating procedure, critical points, acceptance criteria and project management. Both assays were successfully transferred to laboratories that had not performed the assays previously...

  5. Communications standards

    CERN Document Server

    Stokes, A V

    1986-01-01

    Communications Standards deals with the standardization of computer communication networks. This book examines the types of local area networks (LANs) that have been developed and looks at some of the relevant protocols in more detail. The work of Project 802 is briefly discussed, along with a protocol which has developed from one of the LAN standards and is now a de facto standard in one particular area, namely the Manufacturing Automation Protocol (MAP). Factors that affect the usage of networks, such as network management and security, are also considered. This book is divided into three se

  6. Assaying the reporter gene chloramphenicol acetyltransferase

    International Nuclear Information System (INIS)

    Crabb, D.W.; Minth, C.D.; Dixon, J.E.

    1989-01-01

    These experiments document the presence of enzymatic activities in extracts of commonly used cell lines which interfere with the determination of CAT activity. We suspect that the deacetylase activity is the most important, as the extract of the H4IIE C3 cells was capable of completely deacetylating the mono- and diacetylchloramphenicol formed during a 2-hr incubation of CAT with chloramphenicol and acetyl-CoA. The results of the inhibitor experiments are consistent with the presence of proteases which degrade CAT, or a serine carboxylesterase. The interference was also reduced by about half by EDTA; a metalloenzyme (either a protease or esterase) may therefore be involved. This interference appears to be a common phenomenon. We have surveyed 23 different cell types for the presence of the interfering activity and found it in 15. The interference was particularly prominent in several neuroendocrine and hepatoma cells. We took advantage of the effect of EDTA and the heat stability of CAT to eliminate the interference. Addition of 5 mM EDTA and a 10-min incubation of the sonicated cell suspension at 60 degrees prior to centrifugation abolished the interference in all cell lines tested. It is important to note that in order to reveal any CAT activity in some of the extracts (e.g., PC-12 or Hep3B), it was necessary to run the CAT assay for 2 hr. The control assays were therefore run almost to completion, and were well beyond the linear range of the assay. Therefore, the small differences which we observed between the heat-treated and control samples in some instances (e.g., rice, corn, or HeLa cells) will be dramatically amplified when the CAT assay is performed under conditions in which only a small percentage of the substrate is converted to product

  7. A radioreceptor assay for catecholamines, 2

    International Nuclear Information System (INIS)

    Ohmori, Yoshiaki; Nakano, Ryuichi; Lie, Shozo; Imura, Hiroo; Shimbo, Shinichiro.

    1980-01-01

    We have already reported on a radioreceptor assay for catecholamines utilizing the microsome fraction of bovine myocardium as a catecholamine (CA) receptor and 3 H-norepinephrine as a labelled CA. In order to increase the sensitivity of the radioreceptor assay, we used 4-(2-iodoethyl) pyrocatechol ( 125 I-CA) as a ligand instead of 3 H-norepinephrine and performed a radioreceptor assay for CA. The following results were obtained: 1) 125 I-CA was able to bind alpha-receptors prepared from bovine myocardium. 2) The optimal amount of the microsomal fraction was 250 μg/tube, when 125 I-CA of 50,000 c.p.m. was used. The appropriate conditions for incubation were 90 minutes at 20 0 C in a pH 7.0 sucrose solution. 3) By this method utilizing 125 I-CA, norepinephrine was detectable in a range from 500 pg to 10 ng/tube. 4) Various compounds with a catechol nucleus showed cross-reaction in this radioreceptor assay system. 5) Whereas beta-adrenergic blocking agents did not inhibit the binding of 125 I-CA, phentholamine, a short acting type of alpha-adrenergic blocking agents, was effective in inhibiting the binding. However, dibenamine and phenoxybenzamine, long acting types of alpha-adrenergic blocking agents, increased the binding of 125 I-CA to the microsomal fraction. 6) Utilizing this phenomenon, norepinephrine was detectable in the range from 100 pg to 5 ng/tube. (author)

  8. A murine monoclonal antibody based enzyme-linked immunosorbent assay for almond (Prunus dulcis L.) detection.

    Science.gov (United States)

    Su, Mengna; Venkatachalam, Mahesh; Liu, Changqi; Zhang, Ying; Roux, Kenneth H; Sathe, Shridhar K

    2013-11-13

    A sandwich enzyme-linked immunosorbent assay (ELISA) using anti-almond soluble protein rabbit polyclonal antibodies as capture antibodies and murine monoclonal antibody 4C10 as the detection antibodies was developed. The assay is specific and sensitive (3-200 ng almond protein/mL) for almond detection. The standardized assay is accurate (<15% CV) and reproducible (intra- and inter assay variability <15% CV). The assay did not register any cross-reactivity with the tested food matrices, suggesting the assay to be almond amandin specific. The assay could detect the presence of declared almond in the tested matched commercial samples. Further, the assay reliably detected the presence of almonds in the laboratory prepared food samples spiked with almond flour.

  9. In situ assay of nitrate reductase activity using portable water bath.

    Science.gov (United States)

    Rajsz, Adam; Wojtuń, Bronisław; Bytnerowicz, Andrzej

    2017-07-01

    In environmental research (i.e., plant ecophysiology, environmental microbiology, and environmental chemistry), some assays require incubation of samples at controlled temperature and darkness. Until now, due to a lack of equipment providing such possibility in situ, researchers had to move collected samples to the laboratory for incubation. Obviously, a delayed incubation and the ex situ conditions could seriously affect the assays' results. A good example of analysis where water bath use is needed is the nitrate reductase activity (NRA) in vivo assay where plant tissue samples are incubated in buffer solution at a predetermined temperature. We designed a transportable water bath with a temperature control which enables in situ measurements in many types of environmental studies. The presented device is small in size featuring a thermally insulated chamber and an electronically controlled thermostat system powered by a 12-V battery. Due to its modular design, it can be transported comfortably in difficult terrain. The incubation process can be carried out continuously in stable temperature and darkness. In order to examine the field usability of the presented device, we conducted measurements of plant nitrate reductase activity in difficult field conditions. The in situ assays were carried out at high altitudes in the Karkonosze mountains, SW Poland. The NRA was studied in two alpine species (Deschampsia caespitosa and Homogyne alpina). Our results showed low NR activity in H. alpina (mean 0.31 μM NO 2 g -1 DW h -1 ) and higher NRA in D. caespitosa (mean 2.7 μM NO 2 g -1 DW h -1 ). The obtained results were highly reproducible and had small variability (low standard error values).

  10. 21 CFR 864.7375 - Glutathione reductase assay.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Glutathione reductase assay. 864.7375 Section 864.7375 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED... reductase deficiency, or riboflavin deficiency. (b) Classification. Class II (performance standards). [45 FR...

  11. A radioreceptor assay for benzodiazepines

    International Nuclear Information System (INIS)

    Hunt, P.; Husson, J.-M.; Raynaud, J.-P.

    1979-01-01

    A simple, rapid and sensitive radioreceptor assay for determining benzodiazepines in serum is based on the displacement by the drug specific [ 3 H] diazepam binding to a membrane fraction from rat brain. The limit of detection of the more active benzodiazepines is about 0.5 ng. Diazepam, nitrazepam, clobazam and HR 458 have been assayed in human serum after a single oral clinical dose. The results can be used for determining pharmacokinetic parameters. The technique measures not only the parent benzodiazepine but also clinically active metabolites. (author)

  12. Genetic mouse embryo assay: improving performance and quality testing for assisted reproductive technology (ART) with a functional bioassay.

    Science.gov (United States)

    Gilbert, Rebecca S; Nunez, Brandy; Sakurai, Kumi; Fielder, Thomas; Ni, Hsiao-Tzu

    2016-03-24

    Growing concerns about safety of ART on human gametes, embryos, clinical outcomes and long-term health of offspring require improved methods of risk assessment to provide functionally relevant assays for quality control testing and pre-clinical studies prior to clinical implementation. The one-cell mouse embryo assay (MEA) is the most widely used for development and quality testing of human ART products; however, concerns exist due to the insensitivity/variability of this bioassay which lacks standardization and involves subjective analysis by morphology alone rather than functional analysis of the developing embryos. We hypothesized that improvements to MEA by the use of functional molecular biomarkers could enhance sensitivity and improve detection of suboptimal materials/conditions. Fresh one-cell transgenic mouse embryos with green fluorescent protein (GFP) expression driven by Pou6f1 or Cdx2 control elements were harvested and cultured to blastocysts in varied test and control conditions to compare assessment by standard morphology alone versus the added dynamic expression of GFP for screening and selection of critical raw materials and detection of suboptimal culture conditions. Transgenic mouse embryos expressing functionally relevant biomarkers of normal early embryo development can be used to monitor the developmental impact of culture conditions. This novel approach provides a superior MEA that is more meaningful and sensitive for detection of embryotoxicity than morphological assessment alone.

  13. Development of a TaqMan assay for the six major genotypes of hepatitis C virus: Comparison with commercial assays

    DEFF Research Database (Denmark)

    Engle, Ronald E; Russell, Rodney S; Purcell, Robert H

    2008-01-01

    A quantitative real-time PCR assay was developed that detects genomic RNA from reference strains representing the six major genotypes of hepatitis C virus (HCV) with equal sensitivity and accurately measured HCV RNA in JFH1 HCV-infected Huh7.5 cells. The method is indirectly calibrated to the first...... international (WHO 96/790) HCV standard preparation and has a linear dynamic range of 10(2.6)-10(6.5) IU/ml. In addition, the inter- and intra-assay precision were approximately 3% CV and HCV RNA Nucleic Acid Technology kits...... (Versant HCV RNA 3.0 b-DNA and Amplicor HCV Monitor), that also employ the WHO standard, allowed validation of the TaqMan assay against all major HCV genotypes. Both commercial methods detected HCV RNA over a wide dynamic range, but showed a consistent difference of about 0.3 log10 when evaluating samples...

  14. Optimization of protein samples for NMR using thermal shift assays

    Energy Technology Data Exchange (ETDEWEB)

    Kozak, Sandra [European Molecular Biology Laboratory (EMBL), Hamburg Outstation, SPC Facility (Germany); Lercher, Lukas; Karanth, Megha N. [European Molecular Biology Laboratory (EMBL), SCB Unit (Germany); Meijers, Rob [European Molecular Biology Laboratory (EMBL), Hamburg Outstation, SPC Facility (Germany); Carlomagno, Teresa, E-mail: teresa.carlomagno@oci.uni-hannover.de [European Molecular Biology Laboratory (EMBL), SCB Unit (Germany); Boivin, Stephane, E-mail: sboivin77@hotmail.com, E-mail: s.boivin@embl-hamburg.de [European Molecular Biology Laboratory (EMBL), Hamburg Outstation, SPC Facility (Germany)

    2016-04-15

    Maintaining a stable fold for recombinant proteins is challenging, especially when working with highly purified and concentrated samples at temperatures >20 °C. Therefore, it is worthwhile to screen for different buffer components that can stabilize protein samples. Thermal shift assays or ThermoFluor{sup ®} provide a high-throughput screening method to assess the thermal stability of a sample under several conditions simultaneously. Here, we describe a thermal shift assay that is designed to optimize conditions for nuclear magnetic resonance studies, which typically require stable samples at high concentration and ambient (or higher) temperature. We demonstrate that for two challenging proteins, the multicomponent screen helped to identify ingredients that increased protein stability, leading to clear improvements in the quality of the spectra. Thermal shift assays provide an economic and time-efficient method to find optimal conditions for NMR structural studies.

  15. Achieving Standardization

    DEFF Research Database (Denmark)

    Henningsson, Stefan

    2016-01-01

    competitive, national customs and regional economic organizations are seeking to establish a standardized solution for digital reporting of customs data. However, standardization has proven hard to achieve in the socio-technical e-Customs solution. In this chapter, the authors identify and describe what has......International e-Customs is going through a standardization process. Driven by the need to increase control in the trade process to address security challenges stemming from threats of terrorists, diseases, and counterfeit products, and to lower the administrative burdens on traders to stay...... to be harmonized in order for a global company to perceive e-Customs as standardized. In doing so, they contribute an explanation of the challenges associated with using a standardization mechanism for harmonizing socio-technical information systems....

  16. Achieving Standardization

    DEFF Research Database (Denmark)

    Henningsson, Stefan

    2014-01-01

    competitive, national customs and regional economic organizations are seeking to establish a standardized solution for digital reporting of customs data. However, standardization has proven hard to achieve in the socio-technical e-Customs solution. In this chapter, the authors identify and describe what has......International e-Customs is going through a standardization process. Driven by the need to increase control in the trade process to address security challenges stemming from threats of terrorists, diseases, and counterfeit products, and to lower the administrative burdens on traders to stay...... to be harmonized in order for a global company to perceive e-Customs as standardized. In doing so, they contribute an explanation of the challenges associated with using a standardization mechanism for harmonizing socio-technical information systems....

  17. Training Standardization

    International Nuclear Information System (INIS)

    Agnihotri, Newal

    2003-01-01

    The article describes the benefits of and required process and recommendations for implementing the standardization of training in the nuclear power industry in the United States and abroad. Current Information and Communication Technologies (ICT) enable training standardization in the nuclear power industry. The delivery of training through the Internet, Intranet and video over IP will facilitate this standardization and bring multiple benefits to the nuclear power industry worldwide. As the amount of available qualified and experienced professionals decreases because of retirements and fewer nuclear engineering institutions, standardized training will help increase the number of available professionals in the industry. Technology will make it possible to use the experience of retired professionals who may be interested in working part-time from a remote location. Well-planned standardized training will prevent a fragmented approach among utilities, and it will save the industry considerable resources in the long run. It will also ensure cost-effective and safe nuclear power plant operation

  18. Development of real-time reverse transcriptase qPCR assays for the detection of Punta Toro virus and Pichinde virus.

    Science.gov (United States)

    Stefan, Christopher P; Chase, Kitty; Coyne, Susan; Kulesh, David A; Minogue, Timothy D; Koehler, Jeffrey W

    2016-03-31

    Research with high biocontainment pathogens such as Rift Valley fever virus (RVFV) and Lassa virus (LASV) is expensive, potentially hazardous, and limited to select institutions. Surrogate pathogens such as Punta Toro virus (PTV) for RVFV infection and Pichinde virus (PICV) for LASV infection allow research to be performed under more permissive BSL-2 conditions. Although used as infection models, PTV and PICV have no standard real-time RT-qPCR assays to detect and quantify pathogenesis. PTV is also a human pathogen, making a standardized detection assay essential for biosurveillance. Here, we developed and characterized two real-time RT-qPCR assays for PICV and PTV by optimizing assay conditions and measuring the limit of detection (LOD) and performance in multiple clinical matrices. Total nucleic acid from virus-infected Vero E6 cells was used to optimize TaqMan-minor groove binder (MGB) real-time RT-qPCR assays. A 10-fold dilution series of nucleic acid was used to perform analytical experiments with 60 replicates used to confirm assay LODs. Serum and whole blood spiked with 10-fold dilutions of PTV and PICV virus were assessed as matrices in a mock clinical context. The Cq, or cycle at which the fluoresce of each sample first crosses a threshold line, was determined using the second derivative method using Roche LightCycler 480 software version 1.5.1. Digital droplet PCR (ddPCR) was utilized to quantitatively determine RNA target counts/μl for PTV and PICV. Optimized PTV and PICV assays had LODs of 1000 PFU/ml and 100 PFU/ml, respectively, and this LOD was confirmed in 60/60 (PTV) and 58/60 (PICV) positive replicates. Preliminary mock clinical LODs remained consistent in serum and whole blood for PTV and PICV at 1000 PFU/ml and 100 PFU/ml. An exclusivity panel showed no cross reaction with near neighbors. PTV and PICV Taq-man MGB based real-time RT-qPCR assays developed here showed relevant sensitivity and reproducibility in samples extracted from a variety of

  19. Short-term test-retest-reliability of conditioned pain modulation using the cold-heat-pain method in healthy subjects and its correlation to parameters of standardized quantitative sensory testing.

    Science.gov (United States)

    Gehling, Julia; Mainka, Tina; Vollert, Jan; Pogatzki-Zahn, Esther M; Maier, Christoph; Enax-Krumova, Elena K

    2016-08-05

    Conditioned Pain Modulation (CPM) is often used to assess human descending pain inhibition. Nine different studies on the test-retest-reliability of different CPM paradigms have been published, but none of them has investigated the commonly used heat-cold-pain method. The results vary widely and therefore, reliability measures cannot be extrapolated from one CPM paradigm to another. Aim of the present study was to analyse the test-retest-reliability of the common heat-cold-pain method and its correlation to pain thresholds. We tested the short-term test-retest-reliability within 40 ± 19.9 h using a cold-water immersion (10 °C, left hand) as conditioning stimulus (CS) and heat pain (43-49 °C, pain intensity 60 ± 5 on the 101-point numeric rating scale, right forearm) as test stimulus (TS) in 25 healthy right-handed subjects (12females, 31.6 ± 14.1 years). The TS was applied 30s before (TSbefore), during (TSduring) and after (TSafter) the 60s CS. The difference between the pain ratings for TSbefore and TSduring represents the early CPM-effect, between TSbefore and TSafter the late CPM-effect. Quantitative sensory testing (QST, DFNS protocol) was performed on both sessions before the CPM assessment. paired t-tests, Intraclass correlation coefficient (ICC), standard error of measurement (SEM), smallest real difference (SRD), Pearson's correlation, Bland-Altman analysis, significance level p test-retest-reliability of the early CPM-effect using the heat-cold-pain method in healthy subjects achieved satisfying results in terms of the ICC. The SRD of the early CPM effect showed that an individual change of > 20 NRS can be attributed to a real change rather than chance. The late CPM-effect was weaker and not reliable.

  20. semen by MTT reduction assay

    African Journals Online (AJOL)

    user

    the concentration of sperm in each semen sample, sperm motility, plasma integrity of sperm in terms of hypo-osmotic swelling (HOS) test, live and dead ratio of sperm and MTT reduction assay of each ejaculate were determined. Plasma membrane integrity of fresh sperm was assessed using a hypo-osmotic swelling (HOS) ...

  1. Modified Folin-Ciocalteu antioxidant capacity assay for measuring lipophilic antioxidants.

    Science.gov (United States)

    Berker, Kadriye Isil; Ozdemir Olgun, F Ayca; Ozyurt, Dilek; Demirata, Birsen; Apak, Resat

    2013-05-22

    The Folin-Ciocalteu (FC) method of performing a total phenolics assay, originally developed for protein determination, has recently evolved as a total antioxidant capacity assay but was found to be incapable of measuring lipophilic antioxidants due to the high affinity of the FC chromophore, that is, multivalent-charged phospho-tungsto-molybdate(V), toward water. Thus, the FC method was modified and standardized so as to enable simultaneous measurement of lipophilic and hydrophilic antioxidants in NaOH-added isobutanol-water medium. Optimal conditions were as follows: dilution ratio of aqueous FC reagent with iso-BuOH (1:2, v/v), final NaOH concentration of 3.5 × 10(-2) M, reaction time of 20 min, and maximum absorption wavelength of 665 nm. The modified procedure was successfully applied to the total antioxidant capacity assay of trolox, quercetin, ascorbic acid, gallic acid, catechin, caffeic acid, ferulic acid, rosmarinic acid, glutathione, and cysteine, as well as of lipophilic antioxidants such as α-tocopherol (vitamin E), butylated hydroxyanisole, butylated hydroxytoluene, tertiary butylhydroquinone, lauryl gallate, and β-carotene. The modified FC method reliably quantified ascorbic acid, whereas the conventional method could not. The modified method was reproducible and additive in terms of total antioxidant capacity values of constituents of complex mixtures such as olive oil extract and herbal tea infusion. The trolox equivalent antioxidant capacities of the tested antioxidant compounds correlated well with those found by the Cupric Reducing Antioxidant Capacity reference method.

  2. Novel fabrication of immunochromatographic assay based on up conversion phosphors for sensitive detection of clenbuterol.

    Science.gov (United States)

    Wang, Peilong; Wang, Ruiguo; Zhang, Wei; Su, Xiaoou; Luo, Haifeng

    2016-03-15

    A novel and ultra sensitive immunochromatographic assay sensor (ICA) based on up conversion phosphor (UCP) for quantitative detection of clenbuterol (CL) was developed. Monoclonal antibody against CL was labeled with UCP beads. The detection strategy is based on competitive immunoreaction between CL antibodies conjugated to UCP beads and CL or CL antigen on the UCP-ICA sensor. It enables ultra sensitive detection of CL in one single test without complicated sample preparation. Sensing results can be obtained within 10 min. Under optimized conditions, visual limit of detection (vLOD) of UCP-ICA for CL was 0.1 ng/mL. Calculated LOD (cLOD) for CL, as low as 0.01 ng/mL, could be achieved with the UCP-ICA sensor. Recoveries of CL in various sample matrixes ranged from 73.0% to 92.2% and relative standard deviations (RSD) were below 12%. The assay was evaluated with spiked and real samples and the results were compared with liquid chromatography-tandem mass. The developed novel assay method based on UCP could be a potential alternative format for on site and rapid detection of CL as well as other illegal drugs. Copyright © 2015 Elsevier B.V. All rights reserved.

  3. New low-viscosity overlay medium for viral plaque assays

    Directory of Open Access Journals (Sweden)

    Garten Wolfgang

    2006-08-01

    Full Text Available Abstract Background Plaque assays in cell culture monolayers under solid or semisolid overlay media are commonly used for quantification of viruses and antiviral substances. To overcome the pitfalls of known overlays, we tested suspensions of microcrystalline cellulose Avicel RC/CL™ as overlay media in the plaque and plaque-inhibition assay of influenza viruses. Results Significantly larger plaques were formed under Avicel-containing media, as compared to agar and methylcellulose (MC overlay media. The plaque size increased with decreasing Avicel concentration, but even very diluted Avicel overlays (0.3% ensured formation of localized plaques. Due to their low viscosity, Avicel overlays were easier to use than methylcellulose overlays, especially in the 96-well culture plates. Furthermore, Avicel overlay could be applied without prior removal of the virus inoculum thus facilitating the assay and reducing chances of cross-contamination. Using neuraminidase inhibitor oseltamivir carboxylate, we demonstrated applicability of the Avicel-based plaque reduction assay for testing of antiviral substances. Conclusion Plaque assay under Avicel-containing overlay media is easier, faster and more sensitive than assays under agar- and methylcellulose overlays. The assay can be readily performed in a 96-well plate format and seems particularly suitable for high-throughput virus titrations, serological studies and experiments on viral drug sensitivity. It may also facilitate work with highly pathogenic agents performed under hampered conditions of bio-safety labs.

  4. Synthesis and evaluation of phosphorescent oligonucleotide probes for hybridisation assays

    Science.gov (United States)

    O’Sullivan, Paul J.; Burke, Martina; Soini, Aleksi E.; Papkovsky, Dmitri B.

    2002-01-01

    Monofunctional, p-isothiocyanatophenyl-derivatives of platinum (II)-coproporphyrin-I (PtCP-NCS) were evaluated as phosphorescent labelling reagents for synthetic oligonucleotides containing a 3′- or 5′-amino modification. Synthesis and purification conditions were optimised to generate high yields and purity of PtCP-labelled oligonucleotide probes. Phosphorescent properties of the PtCP label have been shown to be largely unaffected by conjugation to oligonucleotides of various length, GC composition and label attachment site. 5′-PtCP-labelled oligonucleotides were shown to work efficiently as primers in a standard PCR. A dedicated 532 nm laser-based time-resolved fluorescence plate reader enabled highly sensitive detection of PtCP-labelled oligonucleotides and PCR products, both in solution and in agarose gels, with limits of detection in the order of 0.3 pM. A model system employing two complementary oligonucleotides labelled with PtCP and QSY® 7 dye (dark quencher) showed strong (∼20-fold) and specific proximity quenching of PtCP label upon hybridisation in solution. The potential applications of PtCP-labelled probes in hybridisation assays were discussed. PMID:12409473

  5. LOCAL SITE CONDITIONS INFLUENCING EARTHQUAKE INTENSITIES AND SECONDARY COLLATERAL IMPACTS IN THE SEA OF MARMARA REGION - Application of Standardized Remote Sensing and GIS-Methods in Detecting Potentially Vulnerable Areas to Earthquakes, Tsunamis and Other Hazards.

    Directory of Open Access Journals (Sweden)

    George Pararas-Carayannis

    2011-01-01

    Full Text Available The destructive earthquake that struck near the Gulf of Izmit along the North Anatolian fault in Northwest Turkey on August 17, 1999, not only generated a local tsunami that was destructive at Golcuk and other coastal cities in the eastern portion of the enclosed Sea of Marmara, but was also responsible for extensive damage from collateral hazards such as subsidence, landslides, ground liquefaction, soil amplifications, compaction and underwater slumping of unconsolidated sediments. This disaster brought attention in the need to identify in this highly populated region, local conditions that enhance earthquake intensities, tsunami run-up and other collateral disaster impacts. The focus of the present study is to illustrate briefly how standardized remote sensing techniques and GIS-methods can help detect areas that are potentially vulnerable, so that disaster mitigation strategies can be implemented more effectively. Apparently, local site conditions exacerbate earthquake intensities and collateral disaster destruction in the Marmara Sea region. However, using remote sensing data, the causal factors can be determined systematically. With proper evaluation of satellite imageries and digital topographic data, specific geomorphologic/topographic settings that enhance disaster impacts can be identified. With a systematic GIS approach - based on Digital Elevation Model (DEM data - geomorphometric parameters that influence the local site conditions can be determined. Digital elevation data, such as SRTM (Shuttle Radar Topography Mission, with 90m spatial resolution and ASTER-data with 30m resolution, interpolated up to 15 m is readily available. Areas with the steepest slopes can be identified from slope gradient maps. Areas with highest curvatures susceptible to landslides can be identified from curvature maps. Coastal areas below the 10 m elevation susceptible to tsunami inundation can be clearly delineated. Height level maps can also help locate

  6. A Non-Isotopic In Vitro Assay for Histone Acetylation

    Science.gov (United States)

    Kuninger, David; Lundblad, James; Semirale, Anthony; Rotwein, Peter

    2007-01-01

    We describe a simple, robust, and relatively inexpensive non-radioactive in vitro assay for measuring histone acetyl-transferase activity. The assay takes advantage of easy to purify recombinant E. coli-derived fusion proteins containing the NH2-terminal tails of histones H3 and H4 linked to epitope-tagged maltose binding protein (MBP), and immunoblotting with antibodies specific to acetylated H3 and H4. Here we show the specificity and dynamic range of this assay for the histone acetyl-transferases, p300 and PCAF. This assay may be adapted readily for other substrates by simply generating new fusion proteins and for other acetyl-transferases by modifying reaction conditions. PMID:17698235

  7. Assay strategies and methods for phospholipases

    International Nuclear Information System (INIS)

    Reynolds, L.J.; Washburn, W.N.; Deems, R.A.; Dennis, E.A.

    1991-01-01

    Of the general considerations discussed, the two issues which are most important in choosing an assay are (1) what sensitivity is required to assay a particular enzyme and (2) whether the assay must be continuous. One can narrow the options further by considering substrate availability, enzyme specificity, assay convenience, or the presence of incompatible side reactions. In addition, the specific preference of a particular phospholipase for polar head group, micellar versus vesicular substrates, and anionic versus nonionic detergents may further restrict the options. Of the many assays described in this chapter, several have limited applicability or serious drawbacks and are not commonly employed. The most commonly used phospholipase assays are the radioactive TLC assay and the pH-stat assay. The TLC assay is probably the most accurate, sensitive assay available. These aspects often outweigh the disadvantages of being discontinuous, tedious, and expensive. The radioactive E. coli assay has become popular recently as an alternative to the TLC assay for the purification of the mammalian nonpancreatic phospholipases. The assay is less time consuming and less expensive than the TLC assay, but it is not appropriate when careful kinetics are required. Where less sensitivity is needed, or when a continuous assay is necessary, the pH-stat assay is often employed. With purified enzymes, when free thiol groups are not present, a spectrophotometric thiol assay can be used. This assay is ∼ as sensitive as the pH-stat assay but is more convenient and more reproducible, although the substrate is not available commercially. Despite the many assay choices available, the search continues for a convenient, generally applicable assay that is both sensitive and continuous

  8. 21 CFR 225.158 - Laboratory assays.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 4 2010-04-01 2010-04-01 false Laboratory assays. 225.158 Section 225.158 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) DRUGS... Laboratory assays. Where the results of laboratory assays of drug components, including assays by State feed...

  9. Standardization: colorfull or dull?

    Science.gov (United States)

    van Nes, Floris L.

    2003-01-01

    After mentioning the necessity of standardization in general, this paper explains how human factors, or ergonomics standardization by ISO and the deployment of information technology were linked. Visual display standardization is the main topic; the present as well as the future situation in this field are treated, mainly from an ISO viewpoint. Some observations are made about the necessary and interesting co-operation between physicists and psychologists, of different nationality, who both may be employed by either private enterprise or governmental institutions, in determining visual display requirements. The display standard that is to succeed the present ISO standards in this area: ISO 9241-3, -7, -8 and ISO 13406-1, -2, will have a scope that is not restricted to office tasks. This means a large extension of the contexts for which display requirements have to be investigated and specified especially if mobile use of displays, under outdoor lighting conditions, is included. The new standard will be structured in such a way that it is better accessible than the present ones for different categories of standards users. The subject color in the new standard is elaborated here. A number of questions are asked as to which requirements on color rendering should be made, taking new research results into account, and how far the new standard should go in making recommendations to the display user.

  10. Reference cells and ploidy in the comet assay

    Directory of Open Access Journals (Sweden)

    Gunnar eBrunborg

    2015-02-01

    Full Text Available In the comet assay, single cells are analyzed with respect to their level of DNA damage. Discrimination of the individual cell or cell type based on DNA content, with concomitant scoring of the DNA damage, is useful since this may allow analysis of mixtures of cells. Different cells can then be characterized based on their ploidy, cell cycle stage, or genome size. We here describe two applications of such a cell type-specific comet assay: (i Testicular cell suspensions, analyzed on the basis of their ploidy during spermatogenesis; and (ii reference cells in the form of fish erythrocytes which can be included as internal standards to correct for inter-assay variations. With standard fluorochromes used in the comet assay, the total staining signal from each cell – whether damaged or undamaged – was found to be associated with the cell’s DNA content. Analysis of the fluorescence intensity of single cells is straightforward since these data are available in scoring systems based on image analysis. The analysis of testicular cell suspensions provides information on cell type specific composition, susceptibility to genotoxicants, and DNA repair. Internal reference cells, either untreated or carrying defined numbers of lesions induced by ionizing radiation, are useful for investigation of experimental factors that can cause variation in comet assay results, and for routine inclusion in experiments to facilitate standardization of methods and comparison of comet assay data obtained in different experiments or in different laboratories. They can also be used - in combination with a reference curve - to quantify the DNA lesions induced by a certain treatment. Fish cells of a range of genome sizes, both greater and smaller than human, are suitable for this purpose and they are inexpensive.

  11. The comet assay in testing the potential genotoxicity of nanomaterials

    Directory of Open Access Journals (Sweden)

    Amaya Azqueta

    2015-06-01

    Full Text Available In the last two decades the production and use of nanomaterials (NMs has impressively increased. Their small size, given a mass equal to that of the corresponding bulk material, implies an increase in the surface area and consequently in the number of atoms that can be reactive. They possess different physical, chemical and biological properties compared to bulk materials of the same composition, which makes them very interesting and valuable for many different applications in technology, energy, construction, electronics, agriculture, optics, paints, textiles, food, cosmetics, medicine... Toxicological assessment of NMs is crucial; the same properties that make them interesting also make them potentially harmful for health and the environment. However, the term NM covers many different kinds of particle , and so there is no simple, standard approach to assessing their toxicity. NMs can enter the cell, interact with cell components and even penetrate the nucleus and interfere with the genetic material. Among the different branches of toxicology, genotoxicity is a main area of concern since it is closely related with the carcinogenic potential of compounds. The Organisation for Economic Co-operation and Development (OECD has published internationally agreed in vitro and in vivo validated test methods to evaluate different genotoxic endpoints of chemicals, including chromosome and gene mutations, and DNA breaks. However not all the assays are suitable to study the genotoxic potential of NMs as has been shown by the OECD Working Party on Manufactured Nanomaterials (WPMN. Moreover, alterations to DNA bases, which are precursors to mutations and of great importance in elucidating the mechanism of action of NMs, are not covered by the OECD guidelines. The in vivo standard comet assay (which measures DNA breaks and alkali-labile sites was included in the OECD assays battery in September 2014 while the in vitro standard comet assay is currently under

  12. Complexometric-spectrophotometric assay of tetracyclines in drug formulations.

    Science.gov (United States)

    Sultan, S M; Alzamil, I Z; Alarfaj, N A

    1988-05-01

    An accurate, rapid and very simple spectrophotometric method for the assay of tetracyclines (tetracycline.HCl, chlorotetracycline.HCl, demeclocycline, oxytetracycline.HCl and doxycycline) has been developed. The method is based on the complexation of iron(III) with tetracyclines in 0.001M sulphuric acid. It has been successfully applied to the assay of tetracyclines in drug formulations, and the interferences of excipients have been examined. The results have been statistically compared with those obtained by two standard methods and found to be very satisfactory.

  13. Total free catecholamines assay by identification of its two functional groups and micro-flow injection chemiluminescence.

    Science.gov (United States)

    Nozaki, O; Kawamoto, H; Moriyama, H

    1999-01-01

    We have developed a novel method of assaying total free catecholamines using sulphuric acid-derivatized beads for extracting and identifying catecholamine (CA) on the surface, and assaying the peroxide produced from CA by chemiluminescence (CL). Current assay methods for CA by electrochemical determination, fluorescence and chemiluminescence need a time-consuming separation by high-performance liquid chromatography. We eliminated this separation step by identifying the two functional groups of CA using a derivatized bead and this resulted in a highly specific CA assay. The principle is as follows: the amino group of CA was trapped by ion binding with a sulphuric acid derivative immobilized on a bead, and the diol of the CA bound to the bead was converted to peroxide with imidazole under alkaline conditions. The peroxide produced was assayed by microflow injection-horseradish peroxidase-catalysed luminol chemiluminescence. We synthesized three types of sulphuric acid-derivative immobilized beads (6.5 mm i.d.). The types of immobilized sulphuric acid derivative used were straight-chain, branched chain and benzenesulphonic, respectively. The order of the three types of beads for extracting CA was: bezenesulphonic type > branched type > straight-chain type. The optimal incubation time for generating peroxide was 30 min. The peroxide generated in the reaction solution was stable with within-run reproducibility of CV 5. 7% after incubation for 80 min. The regression equation of a standard curve for dopamine was Y = 12.8 X(2) + 476X - 373 (where Y = light intensity (RLU), X = concentration of dopamine (micromol/L)). The minimum detection limit of dopamine was 0.1 micromol/L, and the within-run reproducibility of dopamine (10.5 micromol/L) was CV 4.7% (n = 5). This method is applicable to assay of total free CA without use of HPLC. Copyright 1999 John Wiley & Sons, Ltd.

  14. Assay of vitamin B12

    International Nuclear Information System (INIS)

    Tovey, K.C.; Carrick, D.T.

    1982-01-01

    A radioassay is described for vitamin B12 which involves denaturing serum protein binding proteins with alkali. In the denaturation step a dithiopolyol and cyanide are used and in the intrinsic factor assay step a vitamin B12 analogue such as cobinamide is used to bind with any remaining serum proteins. The invention also includes a kit in which the dithiopolyol is provided in admixture with the alkali. The dithiopolyol may be dithiothreitol or dithioerythritol. (author)

  15. Multiplexed assay for protein quantitation in the invertebrate Gammarus fossarum by liquid chromatography coupled to tandem mass spectrometry.

    Science.gov (United States)

    Charnot, Aurore; Gouveia, Duarte; Armengaud, Jean; Almunia, Christine; Chaumot, Arnaud; Lemoine, Jérôme; Geffard, Olivier; Salvador, Arnaud

    2017-06-01

    A highly multiplexed liquid chromatography mass spectrometry-selected reaction monitoring (SRM)-based assay for determination of 40 potential protein biomarkers from Gammarus fossarum, an ecotoxicological relevant species, was described. The assay relies on 71 stable isotope-labeled reported peptide standards for the quantitation of proteins of interest in relation to essential physiological functions such as reproductive cycle, defense mechanism, and enzymes involved in homeostasis process and in energy. A direct linear relationship between the spiked peptide concentration and the area under the peak was clearly demonstrated in biological extracts. Precision and accuracy were determined to be between 1.1 and 21% and between 79 and 120%, respectively, depending on the selected protein in a few samples after optimization of digestion conditions. The validity of the assay was documented for several biomarkers linked with reproduction and the molting process was performed with the assessment of protein levels throughout contrasted physiological process (sex, reproductive status). This assay is easy to use, robust, sensitive, and has high-throughput capabilities. The proposed strategy may be extended to any non-model organisms relevant in environmental science. Graphical abstract ᅟ.

  16. Detection of selected antibiotic resistance genes using multiplex PCR assay in mastitis pathogens in the Czech Republic

    Directory of Open Access Journals (Sweden)

    Vladimir Pyatov

    2017-01-01

    Full Text Available The aim of this research was to develop multiplex polymerase chain reaction assays for the detection of aminoglycoside (strA, strB, sulphonamide (sulI, sulII, tetracycline (tetA, tetB, tetK, tetM, tetO, macrolide and lincosamide (msrA, ermA, ermB, ermC, mefA/E genes of resistance in mastitis pathogens (Escherichia coli, Staphylococcus aureus, Streptococcus uberis, Streptococcus agalactiae and Streptococcus dysgalactiae. Applying the established assays, we investigated the distribution of antibiotic resistance genes in the above mentioned species isolated from milk samples in the Czech Republic. Each assay consisted of seven pairs of primers. Six of them amplified fragments of antibiotic resistance genes and one pair a fragment of a species specific gene. Polymerase chain reaction conditions were optimized to amplify seven gene fragments simultaneously in one reaction. In total, 249 isolates were used, among which 111 were positive for E. coli, 52 for S. aureus and 86 for Streptococcus spp. The majority (60.2% of bacteria carried at least one antibiotic resistance gene and 44.6% were multidrug-resistant. The designed multiplex polymerase chain reaction assays may be applied as diagnostic method to replace or complement standard techniques of antibiotic susceptibility testing in the mentioned pathogens.

  17. Effects of standardized Ginkgo biloba extract on the acquisition, retrieval and extinction of conditioned suppression: Evidence that short-term memory and long-term memory are differentially modulated.

    Science.gov (United States)

    Zamberlam, C R; Vendrasco, N C; Oliveira, D R; Gaiardo, R B; Cerutti, S M

    2016-10-15

    Studies in our laboratory have characterized the putative neuromodulatory effects of a standardized extract of the green leaves of Ginkgo biloba (EGb), which comprises a formulation of 24% ginkgo-flavoglycosides and 6% ginkgo-terpenoid lactones, on conditioned suppression. This model comprises a suitable animal model for investigating the behavioral changes and pharmacological mechanisms that underlie fear memory and anxiety. The characterization of the effects on distinct stages of fear memory or fear extinction will help illustrate both the beneficial and harmful effects. Three hundred adult male Wistar rats were randomly assigned to 30 groups according to the treatment as follows: i-ii) control groups (CS-US and CSno-US); iii) vehicle group (12% Tween®80); and iv-vi) EGb groups (250, 500 and 1000mgkg(-1)); or experimental procedures designed to assess the effects of EGb treatment prior to the acquisition (n=20 per group) and retrieval of conditioned fear (n=10 per group) or prior to the extinction training (n=10 per group) and extinction retention test (n=10 per group). Furthermore, to better understand the effects of acute EGb treatment on fear memory, we conducted two additional analyses: the acquisition of within- and between-session extinction of fear memory (short- and long-term memory, respectively). No difference was identified between the control and treatment groups during the retention test (P>0.05), with the exception of the CSno-US group in relation to all groups (Pmemory, which was verified by the suppression ration in the first trial of extinction training (SR=0.39) and the extinction retention test session (SR=0.53, Pmemory acquisition, which were evaluated during the retention test (SR=0.79). Moreover, EGb administered at 1000mgkg(-1) prior to conditioning did not enhance the long-term extinction memory, i.e., it did not prevent the return of extinguished fear memory in the extinction retention test, in which the spontaneous recovery of fear was

  18. "Life-like" assessment of antimicrobial surfaces by a new touch transfer assay displays strong superiority of a copper alloy compared to silver containing surfaces.

    Directory of Open Access Journals (Sweden)

    Johannes Karl-Mark Knobloch

    Full Text Available Transmission of bacteria from inanimate surfaces in healthcare associated environments is an important source of hospital acquired infections. A number of commercially available medical devices promise to fulfill antibacterial activity to reduce environmental contamination. In this study we developed a touch transfer assay modeling fingerprint transmission to investigate the antibacterial activity of surfaces, with confirmed antibacterial activity by a modified ISO 22196 (JIS Z 2801 assay to test such surfaces under more realistic conditions. Bacteria were taken up from a dry standardized primary contaminated surface (PCS with disinfected fingers or fingers covered with sterile and moistened cotton gloves. Subsequently, bacteria were transferred by pressing on secondary contaminated surfaces (SCS with or without potential antibacterial activity and the relative reduction rate was determined after 24 h. A stable transmission rate between PCS and SCS was observed using moistened sterile gloves. A copper containing alloy displayed at least a tenfold reduction of the bacterial load consistently reaching less than 2.5 cfu/cm2. In contrast, no significant reduction of bacterial contamination by silver containing surfaces and matured pure silver was observed in the touch transfer assay. With the touch transfer assay we successfully established a new reproducible method modeling cross contamination. Using the new method we were able to demonstrate that several surfaces with confirmed antimicrobial activity in a modified ISO 22196 (JIS Z 2801 assay lacked effectiveness under defined ambient conditions. This data indicate that liquid based assays like the ISO 22196 should be critically reviewed before claiming antibacterial activity for surfaces in the setting of contamination of dry surfaces by contact to the human skin. We suggest the newly developed touch transfer assay as a new additional tool for the assessment of potential antimicrobial surfaces

  19. The effect of storage conditions on salivary cortisol concentrations using an enzyme immunoassay

    DEFF Research Database (Denmark)

    Nalla, Anjana A; Thomsen, Gerda; Knudsen, Karen Birgitte Moos

    2015-01-01

    Saliva samples are easy to collect and are applicable for home-sampling, e.g. when studying HPA-axis dynamics to characterize diurnal cortisol profiles and the cortisol awakening response. However, the storing and transport conditions might be critical in the home-sampling approach. Here, we tested...... the stability of saliva cortisol in samples stored at different temperatures and after repeated thawing-freezing cycles when measured with an Enzyme Immuno Assay (EIA). Thirteen healthy volunteers, six women and seven men, mean age 31 (range 26-49) years collected saliva either in the morning hours (08...... it was stored at - 80°C. The last tube was stored directly at - 80°C and served as the 'gold standard'. The saliva samples were assayed using Salivary Cortisol Diagnostic EIA. Differences in cortisol measurements between each of the five conditions and the 'gold standard' (- 80°C) were evaluated by one-sample t...

  20. 40 Years of the Salmonella Mutagenicity Assay: Implications for 21st Century Toxicology

    Science.gov (United States)

    The Salmonella (Ames) mutagenicity assay was developed and introduced by Bruce Ames and colleagues in 1971. Since then, it has become the standard assay for hazard identification of mutagens worldwide. It is a first-tier test for mutagenic activity in the pharmaceutical and chemi...

  1. Standardization of positive controls in diagnostic immunohistochemistry

    DEFF Research Database (Denmark)

    Torlakovic, Emina E; Nielsen, Søren; Francis, Glenn

    2015-01-01

    advances in methodology. An Ad Hoc Expert Committee was formed to address the standardization of controls, which is a missing link in demonstrating and assuring standardization of the various components of dIHC. This committee has also developed a concept of immunohistochemistry critical assay performance...

  2. Frequency standards

    CERN Document Server

    Riehle, Fritz

    2006-01-01

    Of all measurement units, frequency is the one that may be determined with the highest degree of accuracy. It equally allows precise measurements of other physical and technical quantities, whenever they can be measured in terms of frequency.This volume covers the central methods and techniques relevant for frequency standards developed in physics, electronics, quantum electronics, and statistics. After a review of the basic principles, the book looks at the realisation of commonly used components. It then continues with the description and characterisation of important frequency standards

  3. Development of an ATP assay for rapid onboard testing to detect living microorganisms in ballast water

    Science.gov (United States)

    Hyun, Bonggil; Cha, Hyung-Gon; Lee, Nayoung; Yum, Seungshic; Baek, Seung Ho; Shin, Kyoungsoon

    2018-03-01

    Ballast water is a principal pathway for the introduction of pathogens and non-indigenous species to ports worldwide. The International Maritime Organization (IMO) and the United States Coast Guard (USCG) have adopted ballast water management regulations that require, e.g., the installation of shipboard ballast water management systems (BWMS). Rapid and simple analytical methods are needed to monitor whether ballast water disinfection ensures compliance with the discharge standards. In this study laboratory and full scale land-based testing was used to investigate the suitability of an adenosine triphosphate (ATP) assay for quantifying living organisms (≥ 10 and < 50 μm minimum dimension) in ballast water. In laboratory experiments the ATP assay was highly sensitive, with a detection limit of < 5 cells 0.1 mL- 1. Diatom species (Chaetoceros simplex and Skeletonema costatum) had low ATP concentrations compared with dinoflagellate, Raphidophyceae, and Chrysophyceae species. This was because of differences in cell volume, as the ATP concentration increased exponentially with increasing cell volume. Using a regression model between ATP concentration and cell volume, an estimated the pass and fail ATP concentration in this study (788-98,610 pg mL- 1) was developed for the discharge of ballast water. In land-based testing the ATP assay also showed a good correlation with the presence of living natural plankton cells in control samples, but the ATP concentration (137 pg mL- 1) was much lower than the ATP guideline. The low ATP concentration in natural plankton cells may reflect a decline in their biological activity because of extended exposure to dark conditions. Although our results need further validation, the ATP assay is a suitable tool for monitoring compliance of ballast water treatment.

  4. Nest expansion assay: a cancer systems biology approach to in vitro invasion measurements

    Directory of Open Access Journals (Sweden)

    Estrada Lourdes

    2009-07-01

    Full Text Available Abstract Background Traditional in vitro cell invasion assays focus on measuring one cell parameter at a time and are often less than ideal in terms of reproducibility and quantification. Further, many techniques are not suitable for quantifying the advancing margin of collectively migrating cells, arguably the most important area of activity during tumor invasion. We have developed and applied a highly quantitative, standardized, reproducible Nest Expansion Assay (NEA to measure cancer cell invasion in vitro, which builds upon established wound-healing techniques. This assay involves creating uniform circular "nests" of cells within a monolayer of cells using a stabilized, silicone-tipped drill press, and quantifying the margin expansion into an overlaid extracellular matrix (ECM-like component using computer-assisted applications. Findings The NEA was applied to two human-derived breast cell lines, MCF10A and MCF10A-CA1d, which exhibit opposite degrees of tumorigenicity and invasion in vivo. Assays were performed to incorporate various microenvironmental conditions, in order to test their influence on cell behavior and measures. Two types of computer-driven image analysis were performed using Java's freely available ImageJ software and its FracLac plugin to capture nest expansion and fractal dimension, respectively – which are both taken as indicators of invasiveness. Both analyses confirmed that the NEA is highly reproducible, and that the ECM component is key in defining invasive cell behavior. Interestingly, both analyses also detected significant differences between non-invasive and invasive cell lines, across various microenvironments, and over time. Conclusion The spatial nature of the NEA makes its outcome susceptible to the global influence of many cellular parameters at once (e.g., motility, protease secretion, cell-cell adhesion. We propose the NEA as a mid-throughput technique for screening and simultaneous examination of factors

  5. Assays for estimating HIV incidence: updated global market assessment and estimated economic value.

    Science.gov (United States)

    Morrison, Charles S; Homan, Rick; Mack, Natasha; Seepolmuang, Pairin; Averill, Megan; Taylor, Jamilah; Osborn, Jennifer; Dailey, Peter; Parkin, Neil; Ongarello, Stefano; Mastro, Timothy D

    2017-11-01

    Accurate incidence estimates are needed to characterize the HIV epidemic and guide prevention efforts. HIV Incidence assays are cost-effective laboratory assays that provide incidence estimates from cross-sectional surveys. We conducted a global market assessment of HIV incidence assays under three market scenarios and estimated the economic value of improved incidence assays. We interviewed 27 stakeholders, and reviewed journal articles, working group proceedings, and manufacturers' sales figures. We determined HIV incidence assay use in 2014, and estimated use in 2015 to 2017 and in 5 to 10-years under three market scenarios, as well as the cost of conducting national and key population surveys using an HIV incidence assay with improved performance. Global 2014 HIV incidence assay use was 308,900 tests, highest in Asia and mostly for case- and population-based surveillance. Estimated 2015 to 2017 use was 94,475 annually, with declines due to China and the United States discontinuing incidence assay use for domestic surveillance. Annual projected 5 to 10 year use under scenario 1 - no change in technology - was 94,475. For scenario 2 - a moderately improved incidence assay - projected annual use was 286,031. Projected annual use for scenario 3 - game-changing technologies with an HIV incidence assay part of (a) standard confirmatory testing, and (b) standard rapid testing, were 500,000 and 180 million, respectively. As HIV incidence assay precision increases, decreased sample sizes required for incidence estimation resulted in $5 to 23 million annual reductions in survey costs and easily offset the approximately $3 million required to develop a new assay. Improved HIV incidence assays could substantially reduce HIV incidence estimation costs. Continued development of HIV incidence assays with improved performance is required to realize these cost benefits. © 2017 The Authors. Journal of the International AIDS Society published by John Wiley & sons Ltd on

  6. Teachers Voices Interpreting Standards

    Directory of Open Access Journals (Sweden)

    Leo C. Rigsby

    2003-11-01

    Full Text Available The State of Virginia has adopted state-mandated testing that aims to raise the standards of performance for children in our schools in a manner that assigns accountability to schools and to teachers. In this paper we argue that the conditions under which the standards were created and the testing implemented undermine the professionalism of teachers. We believe this result has the further consequence of compromising the critical thinking and learning processes of children. We argue this has happened because teachers’ views and experiences have driven neither the setting of standards nor the assessment of their achievement. We use data from essays by teachers in an innovative masters program to compare teachers’ experiences involving the Virginia Standards of Learning with ideal standards for professional development adopted by the National Board for Professional Teaching Standards. We argue that there are serious negative consequences of the failure to include dialogue with K-12 teachers in setting standards and especially in the creation of assessments to measure performances relative to the standards. We believe the most successful, honest, and morally defensible processes must be built on the experience and wisdom of classroom teachers.

  7. Response definition criteria for ELISPOT assays revisited.

    Science.gov (United States)

    Moodie, Z; Price, L; Gouttefangeas, C; Mander, A; Janetzki, S; Löwer, M; Welters, M J P; Ottensmeier, C; van der Burg, S H; Britten, Cedrik M

    2010-10-01

    No consensus has been reached on how to determine if an immune response has been detected based on raw data from an ELISPOT assay. The goal of this paper is to enable investigators to understand and readily implement currently available methods for response determination. We describe empirical and statistical approaches, identifying the strengths and limitations of each approach to allow readers to rationally select and apply a scientifically sound method appropriate to their specific laboratory setting. Five representative approaches were applied to data sets from the CIMT Immunoguiding Program and the response detection and false positive rates were compared. Simulation studies were also performed to compare empirical and statistical approaches. Based on these, we recommend the use of a non-parametric statistical test. Further, we recommend that six medium control wells or four wells each for both medium control and experimental conditions be performed to increase the sensitivity in detecting a response, that replicates with large variation in spot counts be filtered out, and that positive responses arising from experimental spot counts below the estimated limit of detection be interpreted with caution. Moreover, a web-based user interface was developed to allow easy access to the recommended statistical methods. This interface allows the user to upload data from an ELISPOT assay and obtain an output file of the binary responses.

  8. Assaying environmental nickel toxicity using model nematodes

    Science.gov (United States)

    Rudel, David; Douglas, Chandler; Huffnagle, Ian; Besser, John M.; Ingersoll, Christopher G.

    2013-01-01

    Although nickel exposure results in allergic reactions, respiratory conditions, and cancer in humans and rodents, the ramifications of excess nickel in the environment for animal and human health remain largely undescribed. Nickel and other cationic metals travel through waterways and bind to soils and sediments. To evaluate the potential toxic effects of nickel at environmental contaminant levels (8.9-7,600 µg Ni/g dry weight of sediment and 50-800 µg NiCl2/L of water), we conducted assays using two cosmopolitan nematodes, Caenorhabditis elegans and Pristionchus pacificus. We assayed the effects of both sediment-bound and aqueous nickel upon animal growth, developmental survival, lifespan, and fecundity. Uncontaminated sediments were collected from sites in the Midwestern United States and spiked with a range of nickel concentrations. We found that nickel-spiked sediment substantially impairs both survival from larval to adult stages and adult longevity in a concentration-dependent manner. Further, while aqueous nickel showed no adverse effects on either survivorship or longevity, we observed a significant decrease in fecundity, indicating that aqueous nickel could have a negative impact on nematode physiology. Intriguingly, C. elegansand P. pacificus exhibit similar, but not identical, responses to nickel exposure. Moreover, P. pacificus could be tested successfully in sediments inhospitable to C. elegans. Our results add to a growing body of literature documenting the impact of nickel on animal physiology, and suggest that environmental toxicological studies could gain an advantage by widening their repertoire of nematode species.

  9. Assaying environmental nickel toxicity using model nematodes.

    Directory of Open Access Journals (Sweden)

    David Rudel

    Full Text Available Although nickel exposure results in allergic reactions, respiratory conditions, and cancer in humans and rodents, the ramifications of excess nickel in the environment for animal and human health remain largely undescribed. Nickel and other cationic metals travel through waterways and bind to soils and sediments. To evaluate the potential toxic effects of nickel at environmental contaminant levels (8.9-7,600 µg Ni/g dry weight of sediment and 50-800 µg NiCl2/L of water, we conducted assays using two cosmopolitan nematodes, Caenorhabditis elegans and Pristionchus pacificus. We assayed the effects of both sediment-bound and aqueous nickel upon animal growth, developmental survival, lifespan, and fecundity. Uncontaminated sediments were collected from sites in the Midwestern United States and spiked with a range of nickel concentrations. We found that nickel-spiked sediment substantially impairs both survival from larval to adult stages and adult longevity in a concentration-dependent manner. Further, while aqueous nickel showed no adverse effects on either survivorship or longevity, we observed a significant decrease in fecundity, indicating that aqueous nickel could have a negative impact on nematode physiology. Intriguingly, C. elegans and P. pacificus exhibit similar, but not identical, responses to nickel exposure. Moreover, P. pacificus could be tested successfully in sediments inhospitable to C. elegans. Our results add to a growing body of literature documenting the impact of nickel on animal physiology, and suggest that environmental toxicological studies could gain an advantage by widening their repertoire of nematode species.

  10. Assaying environmental nickel toxicity using model nematodes.

    Science.gov (United States)

    Rudel, David; Douglas, Chandler D; Huffnagle, Ian M; Besser, John M; Ingersoll, Christopher G

    2013-01-01

    Although nickel exposure results in allergic reactions, respiratory conditions, and cancer in humans and rodents, the ramifications of excess nickel in the environment for animal and human health remain largely undescribed. Nickel and other cationic metals travel through waterways and bind to soils and sediments. To evaluate the potential toxic effects of nickel at environmental contaminant levels (8.9-7,600 µg Ni/g dry weight of sediment and 50-800 µg NiCl2/L of water), we conducted assays using two cosmopolitan nematodes, Caenorhabditis elegans and Pristionchus pacificus. We assayed the effects of both sediment-bound and aqueous nickel upon animal growth, developmental survival, lifespan, and fecundity. Uncontaminated sediments were collected from sites in the Midwestern United States and spiked with a range of nickel concentrations. We found that nickel-spiked sediment substantially impairs both survival from larval to adult stages and adult longevity in a concentration-dependent manner. Further, while aqueous nickel showed no adverse effects on either survivorship or longevity, we observed a significant decrease in fecundity, indicating that aqueous nickel could have a negative impact on nematode physiology. Intriguingly, C. elegans and P. pacificus exhibit similar, but not identical, responses to nickel exposure. Moreover, P. pacificus could be tested successfully in sediments inhospitable to C. elegans. Our results add to a growing body of literature documenting the impact of nickel on animal physiology, and suggest that environmental toxicological studies could gain an advantage by widening their repertoire of nematode species.

  11. Position paper on standardization

    International Nuclear Information System (INIS)

    1991-04-01

    The ''NPOC Strategic Plan for Building New Nuclear Plants'' creates a framework within which new standardized nuclear plants may be built. The Strategic Plan is an expression of the nuclear energy industry's serious intent to create the necessary conditions for new plant construction and operation. One of the key elements of the Strategic Plan is a comprehensive industry commitment to standardization: through design certification, combined license, first-of-a-kind engineering, construction, operation and maintenance of nuclear power plants. The NPOC plan proposes four stages of standardization in advanced light water reactors (ALWRs). The first stage is established by the ALWR Utility Requirements Document which specifies owner/operator requirements at a functional level covering all elements of plant design and construction, and many aspects of operations and maintenance. The second stage of standardization is that achieved in the NRC design certification. This certification level includes requirements, design criteria and bases, functional descriptions and performance requirements for systems to assure plant safety. The third stage of standardization, commercial standardization, carries the design to a level of completion beyond that required for design certification to enable the industry to achieve potential increases in efficiency and economy. The final stage of standardization is enhanced standardization beyond design. A standardized approach is being developed in construction practices, operating, maintenance training, and procurement practices. This comprehensive standardization program enables the NRC to proceed with design certification with the confidence that standardization beyond the regulations will be achieved. This confidence should answer the question of design detail required for design certification, and demonstrate that the NRC should require no further regulatory review beyond that required by 10 CFR Part 52

  12. Thyroid Histopathology Assessments for the Amphibian Metamorphosis Assay to Detect Thyroid-active Substances

    Science.gov (United States)

    In support of an Organization for Economic Cooperation and Development (OECD) Amphibian Metamorphosis Assay (AMA) Test Guideline for the detection of substances that interact with the hypothalamic-pituitary-thyroid axis, a document was developed that provides a standardized appro...

  13. Relevant Standards

    Indian Academy of Sciences (India)

    .86: Ethernet over LAPS. Standard in China and India. G.7041: Generic Framing Procedure (GFP). Supports Ethernet as well as other data formats (e.g., Fibre Channel); Protocol of ... IEEE 802.3x for flow control of incoming Ethernet data ...

  14. Kynetic resazurin assay (KRA) for bacterial quantification of foodborne pathogens

    Science.gov (United States)

    Arenas, Yaxal; Mandel, Arkady; Lilge, Lothar

    2012-03-01

    Fast detection of bacterial concentrations is important for the food industry and for healthcare. Early detection of infections and appropriate treatment is essential since, the delay of treatments for bacterial infections tends to be associated with higher mortality rates. In the food industry and in healthcare, standard procedures require the count of colony-forming units in order to quantify bacterial concentrations, however, this method is time consuming and reports require three days to be completed. An alternative is metabolic-colorimetric assays which provide time efficient in vitro bacterial concentrations. A colorimetric assay based on Resazurin was developed as a time kinetic assay (KRA) suitable for bacterial concentration measurements. An optimization was performed by finding excitation and emission wavelengths for fluorescent acquisition. A comparison of two non-related bacteria, foodborne pathogens Escherichia coli and Listeria monocytogenes, was performed in 96 well plates. A metabolic and clonogenic dependence was established for fluorescent kinetic signals.

  15. Development of robotic plasma radiochemical assays for positron emission tomography

    International Nuclear Information System (INIS)

    Alexoff, D.L.; Shea, C.; Fowler, J.S.; Gatley, S.J.; Schlyer, D.J.

    1995-01-01

    A commercial laboratory robot system (Zymate PyTechnology II Laboratory Automation System; Zymark Corporation, Hopkinton, MA) was interfaced to standard and custom laboratory equipment and programmed to perform rapid radiochemical analyses for quantitative PET studies. A Zymark XP robot arm was used to carry out the determination of unchanged (parent) radiotracer in plasma using only solid phase extraction methods. Robotic throughput for the assay of parent radiotracer in plasma is 4--6 samples/hour depending on the radiotracer. Robotic assays of parent compound in plasma were validated for the radiotracers [ 11 C]Benztropine, [ 11 C]cocaine, [ 11 C]clorgyline, [ 11 C]deprenyl, [ 11 C]methadone, [ 11 C]methylphenidate, [ 11 C]raclorpride, and [ 11 C]SR46349B. A simple robot-assisted methods development strategy has been implemented to facilitate the automation of plasma assays of new radiotracers

  16. Insulin radioreceptor assay for human erythrocytes

    International Nuclear Information System (INIS)

    Gambhir, K.K.; Archer, J.A.; Carter, L.

    1977-01-01

    Human erythrocytes have specific insulin receptors. Radioreceptor assay for the determination of insulin binding to these receptors is presented. After two passages over a Boyum-type gradient, erythrocytes from freshly collected heparinized blood were isolated and 3.5 x 10 9 erythrocytes per milliliter were incubated for 2.5 h in a modified pH 8.0 4-(2-hydroxyethyl)-1-piperazine ethane sulfonate buffer, iodinated insulin (80 pg/ml), and a range of unlabeled insulin concentrations(0 to 1 x 10 5 ng/ml). Incubation was terminated by pipetting 200 μl of the incubated suspension onto 200 μl of buffer and 200 μl of dibutyl phthalate in pre-chilled microcentrifuge tubes. After centrifugation, supernatant fluid was aspirated, leaving about 0.1 of the dibutyl phthalate on the cell pellets. Percentage of [ 125 I] insulin bound was determined after radioactivity of the cell pellets was measured in a gamma counter. Under these conditions 11 normal volunteers demonstrated a mean of 7.2 +- 0.44% insulin bound specifically to 3.5 x 10 9 cells. The nonspecific binding varied from 8 to 17% of the total insulin bound. Further, a linear increase of specific binding from 1.35 to 13.55% was observed when the cell concentration was increased from 0.72 to 7.2 x 10 9 cells per milliliter, respectively. Insulin, 100 ng/ml, from several animal species inhibited more than half of the binding of porcine 125 I-labeled insulin. Bovine glucagon inhibited 9.8% and bovine somatotropin inhibited 1.1%, whereas desalanine-desasparagine insulin and human choriogonadotropin (10 int. units) did not inhibit binding of 125 I-labeled insulin. For seven duplicates done on a single assay, the CV was 16.1%, whereas that for 11 assays done on different subjects and on different days was 10.7%. Receptor assays utilizing this technique thus have sufficient specificity and sensitivity to be used for further clinical diagnostic and investigative studies of insulin receptors on human erythrocytes

  17. Detection of Streptococcus pyogenes using rapid visual molecular assay.

    Science.gov (United States)

    Zhao, Xiangna; He, Xiaoming; Li, Huan; Zhao, Jiangtao; Huang, Simo; Liu, Wei; Wei, Xiao; Ding, Yiwei; Wang, Zhaoyan; Zou, Dayang; Wang, Xuesong; Dong, Derong; Yang, Zhan; Yan, Xiabei; Huang, Liuyu; Du, Shuangkui; Yuan, Jing

    2015-09-01

    Streptococcus pyogenes is an increasingly important pathogen in many parts of the world. Rapid and accurate detection of S. pyogenes aids in the control of the infection. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed and validated for the specific detection of S. pyogenes. The assay incorporates two methods: a chromogenic analysis using a calcein/Mn(2+) complex and real-time turbidity monitoring to assess the reaction. Both methods detected the target DNA within 60 min under 64°C isothermal conditions. The assay used specifically designed primers to target spy1258, and correctly identified 111 strains of S. pyogenes and 32 non-S. pyogenes strains, including other species of the genus Streptococcus. Tests using reference strains showed that the LAMP assay was highly specific. The sensitivity of the assay, with a detection limit of 1.49 pg DNA, was 10-fold greater than that of PCR. The LAMP assay established in this study is simple, fast and sensitive, and does not rely upon any special equipment; thus, it could be employed in clinical diagnosis. © FEMS 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  18. Radiosotopic assay and binder therefor

    International Nuclear Information System (INIS)

    Caston, J.D.; Kamen, B.A.

    1976-01-01

    A rapid and less costly radioisotopic assay for measuring the concentration of folate in blood serum is described. This procedure utilizes 3 H-pteroylmonoglutamate, unlabeled 5-methyltetrahydrofolic acid, and a partially purified folate binder, such as for example a folate binder extracted from hog kidney. The procedure involves radioisotopically relating the bound amounts of a labeled folate and a known folate at various concentrations of the known folate in a system containing a predetermined amount of the labeled folate, a predetermined amount of the binder factor for the folates, and a predetermined amount of defolated test serum. 16 claims, 8 drawing figures

  19. One or two serological assay testing strategy for diagnosis of HBV and HCV infection? The use of predictive modelling.

    Science.gov (United States)

    Parry, John V; Easterbrook, Philippa; Sands, Anita R

    2017-11-01

    Initial serological testing for chronic hepatitis B virus (HBV) and hepatitis C virus (HCV) infection is conducted using either rapid diagnostic tests (RDT) or laboratory-based enzyme immunoassays (EIA)s for detection of hepatitis B surface antigen (HBsAg) or antibodies to HCV (anti-HCV), typically on serum or plasma specimens and, for certain RDTs, capillary whole blood. WHO recommends the use of standardized testing strategies - defined as a sequence of one or more assays to maximize testing accuracy while simplifying the testing process and ideally minimizing cost. Our objective was to examine the diagnostic outcomes of a one- versus two-assay serological testing strategy. These data were used to inform recommendations in the 2017 WHO Guidelines on hepatitis B and C testing. Few published studies have compared diagnostic outcomes for one-assay versus two-assay serological testing strategies for HBsAg and anti-HCV. Therefore, the principles of Bayesian statistics were used to conduct a modelling exercise to examine the outcomes of a one-assay versus two-assay testing strategy when applied to a hypothetical population of 10,000 individuals. The resulting model examined the diagnostic outcomes (true and false positive diagnoses; true and false negative diagnoses; positive and negative predictive values as a function of prevalence; and total tests required) for both one-assay and two-assay testing strategies. The performance characteristics assumed for assays used within the testing strategies were informed by WHO prequalification assessment findings and systematic reviews for diagnostic accuracy studies. Each of the presumptive testing strategies (one-assay or two-assay) was modelled at varying prevalences of HBsAg (10%, 2% and 0.4%) and of anti-HCV (40%, 10%, 2% and 0.4%), aimed at representing the range of testing populations typically encountered in WHO Member States. When the two-assay testing strategy was considered, the model assumed the independence of the

  20. A 3-plex methylation assay combined with the FGFR3 mutation assay sensitively detects recurrent bladder cancer in voided urine

    DEFF Research Database (Denmark)

    Kandimalla, Raju; Masius, Roy; Beukers, Willemien

    2013-01-01

    is to determine the sensitivity and specificity of a urine assay for the diagnosis of recurrences in patients with a previous primary NMIBC G1/G2 by using cystoscopy as the reference standard. Experimental Design: We selected eight CpG islands (CGI) methylated in bladder cancer from our earlier genome-wide study......Purpose: DNA methylation is associated with bladder cancer and these modifications could serve as useful biomarkers. FGFR3 mutations are present in 60% to 70% of non–muscle invasive bladder cancer (NMIBC). Low-grade bladder cancer recurs in more than 50% of patients. The aim of this study......, and nonmalignant urines (n = 130). Results: The 3-plex assay identified recurrent bladder cancer in voided urine with a sensitivity of 74% in the validation set. In combination with the FGFR3 mutation assay, a sensitivity of 79% was reached (specificity of 77%). Sensitivity of FGFR3 and cytology was 52% and 57...

  1. Localized irradiations, evaluation through 'Comet Assay'

    International Nuclear Information System (INIS)

    Di Giorgio, Marina; Taja, Maria R.; Nasazzi, Nora B.; Bustos, N.; Cavalieri, H.; Bolgiani, A.

    2000-01-01

    During the last 50 years various radiation accidents involving localized irradiations occurred, resulting mainly from improper handling of sealed sources of Cobalt 60, Cesium 137 or Iridium 192 at work placed for industrial gammagraphy and other radiation sources. Severe skin reaction may developed at the contact sites. Such inhomogeneous irradiations lead to a differential exposure of lymphocytes in lymphatic tissues or other organs that may recirculate into the peripheral blood producing a mixed irradiated and unirradiated population of lymphocytes. Applying the mathematical models 'Contaminated Poisson' of Dolphin and Qdr method of Sasaki, a mean dose in the irradiated body area and its size can be estimated from unstable chromosome aberration scoring. There are also different biophysical techniques that can give response in localized irradiations. Biological dosimetry is a necessary complement to physical and clinical dosimetries. Thus, there is increasing interest in the assessment of biological markers that permit the detection of radiation induced damage in the localized irradiations. The 'Comet Assay' (single cell gel electrophoresis) is a sensitive, rapid and relatively inexpensive method for measuring DNA damage in individual cells. Single cells are embedded in agarose on microscope slides, lysed to remove the majority of the proteins, electrophoresed, then stained with ethidium bromide in order to visualize the DNA. When visualized using a fluorescent microscope, DNA of undamaged cells appears as a spherical mass occupying the cavity formed by the lysed cell. Following radiation damage, the smaller the fragment size and the grater the number of fragments of DNA, the grater the percentage of DNA that it is able to migrate in an electric field, forming a comet image. The assay can be performed under alkaline conditions to examine DNA single strand breaks (SSBs), or in non denaturing (neutral) conditions to measure double strand breaks (DSBs) in individual

  2. A novel microculture kinetic assay (MiCK assay) for malignant cell growth and chemosensitivity.

    Science.gov (United States)

    Kravtsov, V D

    1994-01-01

    The THERMOmax microplate reader was adapted for monitoring the growth kinetics of human leukaemic OCI/AML-2 and mouse tumour J-774.1 cell lines in continuous culture. Fluid evaporation from wells, CO2 escape and contamination were prevented by hermetic sealing of the microcultures in wells of a 96-well microplate, thus enabling the cells to grow exponentially for 72 h under the conditions of the incubated microplate reader. For both OCI/AML-2 cells, which grow in suspension, and adherent J-774.1 cells, a linear correlation was demonstrated between the number of unstained cells seeded in a given microplate well and the optical density (OD) of that well. Therefore, the OD/time curve of the culture could be deemed to be its growth curve. By the use of the linear fit equation, the actual number of the cells in the wells was computable at any time point of the assay. In the chemosensitivity test, an inhibitory effect of ARA-C on the growth of the cells could be estimated by viewing of the growth curves plotted on the screen. The maximum kinetic rates (Vmax) of the curves in the control and the ARA-C-treated wells were compared, yielding a growth inhibition index (GII). Comparison of results of the kinetic chemosensitivity assay with those of a [3H]thymidine incorporation assay revealed that the novel assay is suitable for precise quantitation of the cell chemosensitivity, is more informative and has the added technical advantage of performance without recourse to radioactive or chemically hazardous substances.

  3. Hepatitis B assays in serum, plasma and whole blood on filter paper

    Directory of Open Access Journals (Sweden)

    Mayer Theodor K

    2012-05-01

    Full Text Available Abstract Background Screening and determining the immune status of individuals for hepatitis B is usually done by detecting hepatitis B surface antigen (HBsAg and hepatitis B surface antigen-specific antibodies (HBsAb. In some countries with the highest viral burden, performing these assays is currently impractical. This paper explores the use of filter paper as a blood specimen transport medium. Methods Samples, chosen from routine clinical laboratory pool, were applied and dried onto filter paper. Eluates, from the paper samples, were analyzed as routine clinical specimens on ADVIA Centaur 5634® immunoassay analyzers using the standard HBsAg and HBsAb kits. Dried blood samples were subjected to a range of environmental conditions in order to assess stability. Results After drying and elution the assays showed linearity and precision comparable to clinical assays performed on fresh serum. Elutions at various times during a 149 day incubation period showed very little variability in the Index numbers. All analytes were temperature stable except for a decrease in the HBsAg signal at 42°C. Conclusions Filter paper is an acceptable storage and transport medium for serum to be used in the detection of hepatitis B markers if atmospheric variability can be controlled. HBsAg, HBsAb and HBcAb are all stable for at least five months under storage conditions below room temperature. Drying specimens, particularly serum, on filter paper at remote locations, offers a reasonable solution to the problem of hepatitis surveillance in underdeveloped regions, although some attempt at temperature control might be desirable.

  4. Determining airborne concentrations of spatial repellent chemicals in mosquito behavior assay systems.

    Directory of Open Access Journals (Sweden)

    Nicholas J Martin

    Full Text Available BACKGROUND: Mosquito behavior assays have been used to evaluate the efficacy of vector control interventions to include spatial repellents (SR. Current analytical methods are not optimized to determine short duration concentrations of SR active ingredients (AI in air spaces during entomological evaluations. The aim of this study was to expand on our previous research to further validate a novel air sampling method to detect and quantitate airborne concentrations of a SR under laboratory and field conditions. METHODOLOGY/PRINCIPAL FINDINGS: A thermal desorption (TD gas chromatography-mass spectrometry (GC-MS method was used to determine the amount of dichlorodiphenyltrichloroethane (DDT in samples of air. During laboratory experiments, 1 L volumes of air were collected over 10 min intervals from a three-chamber mosquito behavior assay system. Significantly higher levels of airborne DDT were measured in the chamber containing textiles treated with DDT compared to chambers free of AI. In the field, 57 samples of air were collected from experimental huts with and without DDT for onsite analysis. Airborne DDT was detected in samples collected from treated huts. The mean DDT air concentrations in these two huts over a period of four days with variable ambient temperature were 0.74 µg/m(3 (n = 17; SD = 0.45 and 1.42 µg/m(3 (n = 30; SD = 0.96. CONCLUSIONS/SIGNIFICANCE: The results from laboratory experiments confirmed that significantly different DDT exposure conditions existed in the three-chamber system establishing a chemical gradient to evaluate mosquito deterrency. The TD GC-MS method addresses a need to measure short-term (<1 h SR concentrations in small volume (<100 L samples of air and should be considered for standard evaluation of airborne AI levels in mosquito behavior assay systems. Future studies include the use of TD GC-MS to measure other semi-volatile vector control compounds.

  5. Novel Risk Stratification Assays for Acute Coronary Syndrome.

    Science.gov (United States)

    Ahmed, Haitham M; Hazen, Stanley L

    2017-08-01

    Since identification of aspartate aminotransferase as the first cardiac biomarker in the 1950s, there have been a number of new markers used for myocardial damage detection over the decades. There have also been several generations of troponin assays, each with progressively increasing sensitivity for troponin detection. Accordingly, the "standard of care" for myocardial damage detection continues to change. The purpose of this paper is to review the clinical utility, biological mechanisms, and predictive value of these various biomarkers in contemporary clinical studies. As of this writing, a fifth "next" generation troponin assay has now been cleared by the US Food and Drug Administration for clinical use in the USA for subjects presenting with suspected acute coronary syndromes. Use of these high-sensitivity assays has allowed for earlier detection of myocardial damage as well as greater negative predictive value for infarction after only one or two serial measurements. Recent algorithms utilizing these assays have allowed for more rapid rule-out of myocardial infarction in emergency department settings. In this review, we discuss novel assays available for the risk assessment of subjects presenting with chest pain, including both the "next generation" cardiac troponin assays as well as other novel biomarkers. We review the biological mechanisms for these markers, and explore the positive and negative predictive value of the assays in clinical studies, where reported. We also discuss the potential use of these new markers within the context of future clinical care in the modern era of higher sensitivity troponin testing. Finally, we discuss advances in new platforms (e.g., mass spectrometry) that historically have not been considered for rapid in vitro diagnostic capabilities, but that are taking a larger role in clinical diagnostics, and whose prognostic value and power promise to usher in new markers with potential for future clinical utility in acute coronary

  6. Comparison of functional assays used in the clinical development of a placental malaria vaccine.

    Science.gov (United States)

    Pehrson, Caroline; Heno, Kristine K; Adams, Yvonne; Resende, Mafalda; Mathiesen, Line; Soegaard, Max; de Jongh, Willem A; Theander, Thor G; Salanti, Ali; Nielsen, Morten A

    2017-01-23

    Malaria in pregnancy is associated with significant morbidity in pregnant women and their offspring. Plasmodium falciparum infected erythrocytes (IE) express VAR2CSA that mediates binding to chondroitin sulphate A (CSA) in the placenta. Two VAR2CSA-based vaccines for placental malaria are in clinical development. The purpose of this study was to evaluate the robustness and comparability of binding inhibition assays used in the clinical development of placental malaria vaccines. The ability of sera from animals immunised with different VAR2CSA constructs to inhibit IE binding to CSA was investigated in three in vitro assays using 96-well plates, petri dishes, capillary flow and an ex vivo placental perfusion assay. The inter-assay variation was not uniform between assays and ranged from above ten-fold in the flow assay to two-fold in the perfusion assay. The intra-assay variation was highest in the petri dish assay. A positive correlation between IE binding avidity and the level of binding after antibody inhibition in the petri dish assay indicate that high avidity IE binding is more difficult to inhibit. The highest binding inhibition sensitivity was found in the 96-well and petri dish assays compared to the flow and perfusion assays where binding inhibition required higher antibody titers. The inhibitory capacity of antibodies is not easily translated between assays and the high sensitivity of the 96-well and petri dish assays stresses the need for comparing serial dilutions of serum. Furthermore, IE binding avidity must be in the same range when comparing data from different days. There was an overall concordance in the capacity of antibody-mediated inhibition, when comparing the in vitro assays with the perfusion assay, which more closely represents in vivo conditions. Importantly the ID1-ID2a protein in a liposomal formulation, currently in a phase I trial, effectively induced antibodies that inhibited IE adhesion in placental tissue. Copyright © 2016

  7. ImmunoCAP assays: Pros and cons in allergology.

    Science.gov (United States)

    van Hage, Marianne; Hamsten, Carl; Valenta, Rudolf

    2017-10-01

    Allergen-specific IgE measurements and the clinical history are the cornerstones of allergy diagnosis. During the past decades, both characterization and standardization of allergen extracts and assay technology have improved. Here we discuss the uses, advantages, misinterpretations, and limitations of ImmunoCAP IgE assays (Thermo Fisher Scientific/Phadia, Uppsala, Sweden) in the field of allergology. They can be performed as singleplex (ImmunoCAP) and, for the last decade, as multiplex (Immuno Solid-phase Allergen Chip [ISAC]). The major benefit of ImmunoCAP is the obtained quantified allergen-specific IgE antibody level and the lack of interference from allergen-specific IgG antibodies. However, ImmunoCAP allergen extracts are limited to the composition of the extract. The introduction of allergen molecules has had a major effect on analytic specificity and allergy diagnosis. They are used in both singleplex ImmunoCAP and multiplex ImmunoCAP ISAC assays. The major advantage of ISAC is the comprehensive IgE pattern obtained with a minute amount of serum. The shortcomings are its semiquantitative measurements, lower linear range, and cost per assay. With respect to assay performance, ImmunoCAP allergen extracts are good screening tools, but allergen molecules dissect the IgE response on a molecular level and put allergy research on the map of precision medicine. Copyright © 2017 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

  8. Inflammatory markers in Huntington's disease plasma—A robust nanoLC–MRM-MS assay development

    Directory of Open Access Journals (Sweden)

    Melinda Rezeli

    2014-06-01

    Full Text Available The development of an MRM assay for the measurements of six inflammatory markers is presented. We report a robust and sensitive quantitative assay with a relative standard deviation of <15% that accounts for the entire sample processing. The assay has a dynamic range with 4 orders of magnitude and the LOQs are in the attomolar range. We used plasma from Huntington's disease gene carriers and healthy controls to compare our MRM method with antibody based methods. Importantly, we found a good agreement between assays for the measurement of C-reactive protein, in contrast to complement component 3 and complement factor H.

  9. Nondestructive assay of plutonium residue in horizontal storage tanks

    International Nuclear Information System (INIS)

    Marsh, S.F.

    1985-01-01

    Aqueous plutonium recovery and purification processes often involve the temporary storage of plutonium solutions in holding tanks. Because plutonium is known to precipitate from aqueous solutions under certain conditions, there is a continuing need to assay emptied tanks for plutonium residue. A portable gamma spectrometer system, specifically designed for this purpose, provides rapid assay of such plutonium residues in horizontal storage tanks. A means is thus available for the nondestructive analysis of these tanks on a regular schedule to ensure that significant deposits of plutonium are not allowed to accumulate. 5 figs

  10. Conditionals in Interaction

    Directory of Open Access Journals (Sweden)

    Trafford James

    2017-03-01

    Full Text Available There are several issues with the standard approach to the relationship between conditionals and assertions, particularly when the antecedent of a conditional is (or may be false. One prominent alternative is to say that conditionals do not express propositions, but rather make conditional assertions that may generate categorical assertions of the consequent in certain circumstances. However, this view has consequences that jar with standard interpretations of the relationship between proofs and assertion. Here, I analyse this relationship, and say that, on at least one understanding of proof, conditional assertions may reflect the dynamics of proving, which (sometimes generate categorical assertions. In particular, when we think about the relationship between assertion and proof as rooted in a dialogical approach to both, the distinction between conditional and categorical assertions is quite natural.

  11. Microcoupon Assay Of Adhesion And Growth Of Bacterial Films

    Science.gov (United States)

    Pierson, Duane L.; Koenig, David W.

    1994-01-01

    Microbiological assay technique facilitates determination of some characteristics of sessile bacteria like those that attach to and coat interior walls of water-purification systems. Biofilms cause sickness and interfere with purification process. Technique enables direct measurement of rate of attachment of bacterial cells, their metabolism, and effects of chemicals on them. Used to quantify effects of both bactericides and growth-stimulating agents and in place of older standard plate-count and tube-dilution techniques.

  12. Capillary Electrophoresis Analysis of Conventional Splicing Assays

    DEFF Research Database (Denmark)

    de Garibay, Gorka Ruiz; Acedo, Alberto; García-Casado, Zaida

    2014-01-01

    of these assays is often challenging. Here, we explore this issue by conducting splicing assays in 31 BRCA2 genetic variants. All variants were assessed by RT-PCR followed by capillary electrophoresis and direct sequencing. If assays did not produce clear-cut outputs (Class-2 or Class-5 according to analytical...

  13. Multicentre comparison of a diagnostic assay

    DEFF Research Database (Denmark)

    Waters, Patrick; Reindl, Markus; Saiz, Albert

    2016-01-01

    ) assays in neuromyelitis optica spectrum disorders (NMOSD). METHODS: Coded samples from patients with neuromyelitis optica (NMO) or NMOSD (101) and controls (92) were tested at 15 European diagnostic centres using 21 assays including live (n=3) or fixed cell-based assays (n=10), flow cytometry (n=4...

  14. Fast facility spent-fuel and waste assay instrument. [Fluorinel Dissolution and Fuel Storage (FAST) Facility

    Energy Technology Data Exchange (ETDEWEB)

    Eccleston, G.W.; Johnson, S.S.; Menlove, H.O.; Van Lyssel, T.; Black, D.; Carlson, B.; Decker, L.; Echo, M.W.

    1983-01-01

    A delayed-neutron assay instrument was installed in the Fluorinel Dissolution and Fuel Storage Facility at Idaho National Engineering Laboratory. The dual-assay instrument is designed to measure both spent fuel and waste solids that are produced from fuel processing. A set of waste standards, fabricated by Los Alamos using uranium supplied by Exxon Nuclear Idaho Company, was used to calibrate the small-sample assay region of the instrument. Performance testing was completed before installation of the instrument to determine the effects of uranium enrichment, hydrogenous materials, and neutron poisons on assays. The unit was designed to measure high-enriched uranium samples in the presence of large neutron backgrounds. Measurements indicate that the system can assay low-enriched uranium samples with moderate backgrounds if calibrated with proper standards.

  15. An investigation into the suitability of a commercial real-time PCR assay to screen for Taylorella equigenitalis in routine prebreeding equine genital swabs.

    Science.gov (United States)

    Ousey, J C; Palmer, L; Cash, R S G; Grimes, K J; Fletcher, A P; Barrelet, A; Foote, A K; Manning, F M; Ricketts, S W

    2009-12-01

    Standard bacteriological methods for identifying Taylorella equigenitalis in cervical smears are time consuming. Therefore, a more rapid real-time PCR assay was evaluated for its suitability in screening swabs. To compare the results of a commercially available real-time PCR assay with routine microbiological culture for the identification of T. equigenitalis, the causative organism of contagious equine metritis, in equine genital swab samples, under 'field trial' conditions. Routine prebreeding genital swabs (n=2072) collected from Thoroughbred mares and stallions during 2009 were examined together with stored T. equigenitalis positive material. Swabs were cultured for T. equigenitalis using standard microbiological techniques. Bacterial lysates were isolated from the swabs and examined for the presence of a 16S DNA fragment of T. equigenitalis, using a commercial multiplex real-time PCR assay system. There was complete concordance between positive and negative results obtained by the 2 methods. Real-time PCR also detected T. equigenitalis DNA from swabs that were negative using standard microbiological culture after 6 months' storage at +4 degrees C but from which T. equigenitalis had been isolated following collection. The sensitivities of real-time PCR and bacterial culture were both 10(-3) (equivalent to 3 colony-forming units). Routine bacterial culture of T. equigenitalis requires an incubation period of not less than 7 days before a conclusive negative result can be obtained, whereas bacterial extraction and real-time PCR assay can be completed in less than 6 h. The commercially-available PCR assay tested provided a rapid and reliable method for the identification of T. equigenitalis from equine genital swabs and could be usefully employed for the screening of mares and stallions for preseason Horserace Betting Levy Board (HBLB) Code of Practice and in other situations such as for bloodstock sales screening requirements, overcoming the current delays

  16. Non destructive assay (NDA) techniques

    International Nuclear Information System (INIS)

    Mafra Guidicini, Olga; Llacer, Carlos D.; Rojo, Marcelo

    2001-01-01

    In the IAEA Safeguards System the basic verification method used is nuclear material accountancy, with containment and surveillance as important complementary measures. If nuclear material accountancy is to be effective, IAEA inspectors have to make independent measurements to verify declared material quantities. Most of the equipment available to the inspectors is designed to measure gamma rays and/or neutrons emitted by various nuclear materials. Equipment is also available to measure the gross weight of an item containing nuclear material. These types of measurement techniques are generally grouped under the title of nondestructive assay (NDA). The paper describes the NDA techniques and instruments used to verify the total amount of nuclear material held at a nuclear facility. (author)

  17. Development of glomerulus-, tubule-, and collecting duct-specific mRNA assay in human urinary exosomes and microvesicles.

    Directory of Open Access Journals (Sweden)

    Taku Murakami

    Full Text Available Urinary exosomes and microvesicles (EMV are promising biomarkers for renal diseases. Although the density of EMV is very low in urine, large quantity of urine can be easily obtained. In order to analyze urinary EMV mRNA, a unique filter device to adsorb urinary EMV from 10 mL urine was developed, which is far more convenient than the standard ultracentrifugation protocol. The filter part of the device is detachable and aligned to a 96-well microplate format, therefore multiple samples can be processed simultaneously in a high throughput manner following the isolation step. For EMV mRNA quantification, the EMV on the filter is lysed directly by adding lysis buffer and transferred to an oligo(dT-immobilized microplate for mRNA isolation followed by cDNA synthesis and real-time PCR. Under the optimized assay condition, our method provided comparable or even superior results to the standard ultracentrifugation method in terms of mRNA assay sensitivity, linearity, intra-assay reproducibility, and ease of use. The assay system was applied to quantification of kidney-specific mRNAs such as NPHN and PDCN (glomerular filtration, SLC12A1 (tubular absorption, UMOD and ALB (tubular secretion, and AQP2 (collecting duct water absorption. 12-hour urine samples were collected from four healthy subjects for two weeks, and day-to-day and individual-to-individual variations were investigated. Kidney-specific genes as well as control genes (GAPDH, ACTB, etc. were successfully detected and confirmed their stable expressions through the two-week study period. In conclusion, this method is readily available to clinical studies of kidney diseases.

  18. Making transuranic assay measurements using modern controllers

    International Nuclear Information System (INIS)

    Kuckertz, T.H.; Caldwell, J.T.; Medvick, P.A.; Kunz, W.E.; Hastings, R.D.

    1987-01-01

    This paper describes methodology and computer-controlled instrumentation developed at the Los Alamos National Laboratory that accurately performs nondestructive assays of large containers bearing transuranic wastes and nonradioactive matrix materials. These assay systems can measure fissile isotopes with 1-mg sensitivity and spontaneous neutron-emitting isotopes at a 10-mg sensitivity. The assays are performed by neutron interrogation, detection, and counting in a custom assay chamber. An International Business Machines Personal Computer (IBM-PC) is used to control the CAMAC-based instrumentation system that acquires the assay data. 6 refs., 7 figs

  19. Functional assay of the alternative complement pathway of rat serum

    International Nuclear Information System (INIS)

    Coonrod, J.D.; Jenkins, S.D.

    1979-01-01

    Two functional assays of the alternative pathway of complement activation in rat serum were developed. In the first assay, conditions were established for titration of alternative pathway activity by use of the 50% hemolytic end-point of rabbit red blood cells (RaRBC) in serum treated with ethyleneglycol-bis-(beta-aminoethyl ether)-N, N'-tetraacetic acid (EGTA). The second assay of alternative pathway activity was based on the opsonization of heat-killed radiolabeled pneumococci of serotype 25 (Pn25). Opsonization of Pn25 was shown to proceed entirely via the alternative pathway in rat serum. There was excellent correlation between the results obtained with the RaRBC lysis test and those obtained with the opsonization test. Because of its technical simplicity, the RaRBC lysis test appeared to be the single most useful test of alternative pathway activity in rat serum. (Auth.)

  20. QA Objectives for NDA with the Residues Segmented Gamma Scan Assay System at the Plutonium Finishing Plant (PFP)

    International Nuclear Information System (INIS)

    WESTSIK, G.A.

    2001-01-01

    The PFP facility utilizes a Segmented Gamma Scanner Assay System (SGSAS) to perform assays on cans of ash for WIPP characterization measurements. This report documents the conformance of SGSAS to the precision and accuracy radioassay QAOs, and reports the minimum detectable concentration (MDC). The QAO measurement runs supplied in this document were for a billet can geometry. The measurements were performed in August 2000. This document covers assays performed until October 27, 2000. The billet cans containing stabilized residues will be loaded into pipe overpack containers (POC) for shipment to WIPP. The WIPP-WAC defines four nominal test levels for NDA, which are in alpha curies and grams of weapons grade (WG) Pu. Due to intended utilization of the SGSAS system for the materials mentioned above, it is presently only being qualified for the two highest QAO ranges. The sources used for the QAO measurements are plutonium standards, which have been calibrated using calorimetry techniques. This report documents the analysis of test data for the SGSAS system at the nominal 10 gram and 160 gram levels. The MDC was determined using a billet can filled with diatomaceous earth but no plutonium present. Since the system is not being qualified for TRU vs low-level waste (LLW) sorting the MDC will primarily provide verification that the detection level for the system is well below the QAO ranges for which the system is being qualified. The MDC reflects the best sensitivity for a particular assay system and specific assay conditions (i.e. count time, sample configuration) when no added radioactivity is present. As such, no radioactive sources were required for the MDC determination. As with the accuracy and precision QAOs, the MDC is valid for the billet cans

  1. Antibiotic microbial assay using kinetic-reading microplate system

    Directory of Open Access Journals (Sweden)

    Felipe Rebello Lourenço

    2011-09-01

    Full Text Available The aim of this study was to determine the optimal experimental conditions to develop a methodology for microbiological assay of apramycin employing microplate and kinetic reading mode, and to validate the developed method, through evaluation of parameters of selectivity, linearity, linear range, limits of detection and quantification, accuracy and precision. The turbidimetric assay principle is simple: the test solution is added to a suspension of test microorganism in culture media, the mixture is incubated under appropriate conditions and the microbial growth is measured by photometric reading. Microplate with kinetic reading mode employed in antibiotic assay is of considerable interest since it allows reduction of material and analysis time and enables a large number of samples to be analyzed simultaneously, with automated reading and calculating. Established conditions considered the standard-curve of apramycin at concentrations from 5.0 to 35.0 μg mL-1, and tryptic soy broth inoculated with 5% Escherichia coli (ATCC 8739 suspension. Satisfactory results were obtained with 2 hours of incubation. The developed method showed appropriate selectivity, linearity in the range from 5.0 to 35.0 μg mL-1, limits of detection and quantification of 0.1 and 0.4 μg mL-1, respectively, as well as satisfactory accuracy (recuperation = 98.5% and precision (RSD = 6.0%. Microplate assay combined the characteristics of microbiological (evaluation of antibiotic activity against sensitive test microorganism and physico-chemical (operationally straightforward and faster results assays.O objetivo deste trabalho é determinar as condições experimentais ideais para o desenvolvimento de metodologia para a dosagem microbiológica de apramicina empregando microplacas e modo de leitura cinético e validar o método desenvolvido, através da avaliação dos parâmetros de especificidade e seletividade, linearidade, faixa ou intervalo linear, limite de detecção e

  2. Novel interferon-gamma assays for diagnosing tuberculosis in young children in India

    Science.gov (United States)

    Shaikh, N.; Gupte, A.; Dharmshale, S.; Pokkali, S.; Thakar, M.; Upadhye, V. J.; Ordonez, A. A.; Kinikar, A.; Gupte, N.; Mave, V.; Kagal, A.; Gupta, A.; Lalvani, A.; Paranjpe, R.; Bharadwaj, R.; Jain, S. K.

    2017-01-01

    SUMMARY SETTING The tuberculin skin test (TST) and interferon-gamma release assays (IGRAs) are used as supportive evidence to diagnose active tuberculosis (TB). Novel IGRAs could improve diagnosis, but data are lacking in young children. DESIGN Children (age ≤ 5 years) with suspected TB were prospectively screened at a tertiary hospital in Pune, India; the children underwent TST, and standard (early secretory antigenic target 6 and culture filtrate protein 10) and enhanced (five additional novel antigens) enzyme-linked immunospot (ELISpot) assays. RESULTS Of 313 children (median age 30 months) enrolled, 92% had received bacille Calmette-Guérin vaccination, 53% were malnourished and 9% were coinfected with the human immunodeficiency virus (HIV); 48 (15%) had TB, 128 (41%) did not, and TB could not be ruled out in 137 (44%). The sensitivity of enhanced (45%) and standard (42%) ELISpot assays for diagnosing TB was better than that of TST (20%) (P ≤ 0.03); however, enhanced ELISpot was not more sensitive than the standard ELISpot assay (P= 0.50). The specificity of enhanced ELISpot, standard ELISpot and TST was respectively 82% (95%CI 74–89), 88% (95%CI 81–94) and 98% (95%CI 93–100). Rv3879c and Rv3615c, previously reported to be promising antigens, failed to improve the diagnostic performance of the ELISpot assay. CONCLUSION The TST and the standard and novel ELISpot assays performed poorly in diagnosing active TB among young children in India. PMID:28284256

  3. Oxytetracycline Assay in Pond Sediment

    OpenAIRE

    L. Nepejchalová; Z. Svobodová; J. Kolářová; K. Frgalová; J. Valová; D. Némethová

    2008-01-01

    The fate of drug residues and their metabolites in the environment is relatively rarely investigated in the conditions of the Czech Republic, resulting in limited availability of scientific information. To demonstrate one example, we prepared a model study with medicated feedstuff containing oxytetracycline hydrochloride (OTC HCl), which was used in fish under normal conditions of use. The oxytetracycline (OTC) contents were determined in the sediments of the pond where the fish were treated....

  4. A novel in vitro assay for murine haematopoietic stem cells.

    Science.gov (United States)

    Eckmann, L; Freshney, M; Wright, E G; Sproul, A; Wilkie, N; Pragnell, I B

    1988-12-01

    Study of the biology of haematopoietic stem cells is crucially dependent on the availability of suitable in vitro assays. Existing assays have suffered from the fact that they detect small subcompartments of the total stem cell compartment. This limits experiments where it is required to assay a high proportion of stem cells, e.g. the enumeration of stem cell numbers under varying conditions or the identification and purification of stem cell regulators. We describe an in vitro assay which shows macroscopic colony formation and limited self-renewal capacity in vitro. The detected cell (CFU-A) has a low cycling status in normal bone marrow (NBM) and responds to known stem cell regulators. The incidence (100-200 per 10(5) in NBM), the proliferative characteristics under stress and some of the physical properties are similar to stem cells detected by colony formation after transplantation into lethally irradiated recipients (CFU-S). These data indicate that our assay detects a high proportion of haematopoietic stem cells in vitro. This will facilitate experiments on stem cell behaviour which have previously been difficult to conduct.

  5. Irradiation detection of food by DNA Comet Assay

    International Nuclear Information System (INIS)

    Khan, A.A.; Delincee, H.

    1999-01-01

    Microgel electrophoresis of single cells or nuclei (DNA Comet Assay) has been investigated to detect irradiation treatment of more than 50 food commodities e.g. meats, seafood, cereals, pulses, nuts, fruits and vegetables, and spices. The foodstuffs have been exposed to radiation doses covering the range of potential commercial irradiation for inactivation of pathogenic and spoilage micro-organisms, for insect disinfestation and for shelf-life extension. The Comet Assay is based on detection of DNA fragments presumptive to irradiation. For most of the food items investigated, the assay can be applied successfully for irradiation detection by working out different conditions of the assay. However, with some of the foods difficulties arose due to - lack of discrimination between the irradiated and unirradiated food samples due to the presence of the same kinds of comets in both cases and the total absence of the typical intact cells in unirradiated samples. - Sufficient DNA material was not available from some of the foods. - Insufficient lysis of the cell walls in case of some plant foods. In conclusion, the DNA Comet Assay can help to detect the irradiation treatment of several varieties of foods using low-cost equipment in a short time of analysis. (orig.)

  6. Widespread nanoparticle-assay interference: implications for nanotoxicity testing.

    Science.gov (United States)

    Ong, Kimberly J; MacCormack, Tyson J; Clark, Rhett J; Ede, James D; Ortega, Van A; Felix, Lindsey C; Dang, Michael K M; Ma, Guibin; Fenniri, Hicham; Veinot, Jonathan G C; Goss, Greg G

    2014-01-01

    The evaluation of engineered nanomaterial safety has been hindered by conflicting reports demonstrating differential degrees of toxicity with the same nanoparticles. The unique properties of these materials increase the likelihood that they will interfere with analytical techniques, which may contribute to this phenomenon. We tested the potential for: 1) nanoparticle intrinsic fluorescence/absorbance, 2) interactions between nanoparticles and assay components, and 3) the effects of adding both nanoparticles and analytes to an assay, to interfere with the accurate assessment of toxicity. Silicon, cadmium selenide, titanium dioxide, and helical rosette nanotubes each affected at least one of the six assays tested, resulting in either substantial over- or under-estimations of toxicity. Simulation of realistic assay conditions revealed that interference could not be predicted solely by interactions between nanoparticles and assay components. Moreover, the nature and degree of interference cannot be predicted solely based on our current understanding of nanomaterial behaviour. A literature survey indicated that ca. 95% of papers from 2010 using biochemical techniques to assess nanotoxicity did not account for potential interference of nanoparticles, and this number had not substantially improved in 2012. We provide guidance on avoiding and/or controlling for such interference to improve the accuracy of nanotoxicity assessments.

  7. Widespread nanoparticle-assay interference: implications for nanotoxicity testing.

    Directory of Open Access Journals (Sweden)

    Kimberly J Ong

    Full Text Available The evaluation of engineered nanomaterial safety has been hindered by conflicting reports demonstrating differential degrees of toxicity with the same nanoparticles. The unique properties of these materials increase the likelihood that they will interfere with analytical techniques, which may contribute to this phenomenon. We tested the potential for: 1 nanoparticle intrinsic fluorescence/absorbance, 2 interactions between nanoparticles and assay components, and 3 the effects of adding both nanoparticles and analytes to an assay, to interfere with the accurate assessment of toxicity. Silicon, cadmium selenide, titanium dioxide, and helical rosette nanotubes each affected at least one of the six assays tested, resulting in either substantial over- or under-estimations of toxicity. Simulation of realistic assay conditions revealed that interference could not be predicted solely by interactions between nanoparticles and assay components. Moreover, the nature and degree of interference cannot be predicted solely based on our current understanding of nanomaterial behaviour. A literature survey indicated that ca. 95% of papers from 2010 using biochemical techniques to assess nanotoxicity did not account for potential interference of nanoparticles, and this number had not substantially improved in 2012. We provide guidance on avoiding and/or controlling for such interference to improve the accuracy of nanotoxicity assessments.

  8. Optimization of the T-cell proliferation assay in fascioliasis using a ...

    African Journals Online (AJOL)

    T-cell proliferation studies are traditionally carried out with radioactive reagents or fluorescent reagents that require measurement with advanced technology instrumentation. We attempted to calibrate the optimal conditions suitable for the use of a non-radioactive assay for the measurement of a T-cell proliferation assay in ...

  9. A dual immunocytochemical assay for oestrogen and epidermal growth factor receptors in tumour cell lines

    NARCIS (Netherlands)

    A.K. Sharma (Anisha K.); J.H. Horgan; R.L. McClelland (Robyn); A.G. Douglas-Jones (A.); T. van Agthoven (Ton); L.C.J. Dorssers (Lambert); R.I. Nicholson (R.)

    1994-01-01

    textabstractA new dual immunocytochemical assay for oestrogen receptor (ER) and epidermal growth factor receptor (EGFR) has been developed. It has been tested in a variety of conditions using cell culture lines and the results correlate well with those obtained from single immunocytochemical assays.

  10. Human anti-animal antibody interferences in immunological assays.

    Science.gov (United States)

    Kricka, L J

    1999-07-01

    The scope and significance of human anti-animal antibody interference in immunological assays is reviewed with an emphasis on human anti-animal immunoglobulins, particularly human anti-mouse antibodies (HAMAs). Anti-animal antibodies (IgG, IgA, IgM, IgE class, anti-isotype, and anti-idiotype specificity) arise as a result of iatrogenic and noniatrogenic causes and include human anti-mouse, -rabbit, -goat, -sheep, -cow, -pig, -rat, and -horse antibodies and antibodies with mixed specificity. Circulating antibodies can reach gram per liter concentrations and may persist for years. Prevalence estimates for anti-animal antibodies in the general population vary widely and range from HAMA, which causes both positive and negative interferences in two-site mouse monoclonal antibody-based assays. Strategies to prevent the development of human anti-animal antibody responses include immunosuppressant therapy and the use of humanized, polyethylene glycolylated, or Fab fragments of antibody agents. Sample pretreatment or assay redesign can eliminate immunoassay interferences caused by anti-animal antibodies. Enzyme immunoassays, immunoradiometric assays, immunofluorescence, and HPLC assays have been designed to detect HAMA and other anti-animal antibodies, but intermethod comparability is complicated by differences in assay specificity and lack of standardization. Human anti-animal antibodies often go unnoticed, to the detriment of patient care. A heightened awareness on the part of laboratory staff and clinicians of the problems caused by this type of interference in routine immunoassay tests is desirable. Efforts should be directed at improving methods for identifying and eliminating this type of analytical interference.

  11. Standardization of depression measurement

    DEFF Research Database (Denmark)

    Wahl, Inka; Löwe, Bernd; Bjørner, Jakob

    2014-01-01

    OBJECTIVES: To provide a standardized metric for the assessment of depression severity to enable comparability among results of established depression measures. STUDY DESIGN AND SETTING: A common metric for 11 depression questionnaires was developed applying item response theory (IRT) methods. Data...... of 33,844 adults were used for secondary analysis including routine assessments of 23,817 in- and outpatients with mental and/or medical conditions (46% with depressive disorders) and a general population sample of 10,027 randomly selected participants from three representative German household surveys....... RESULTS: A standardized metric for depression severity was defined by 143 items, and scores were normed to a general population mean of 50 (standard deviation = 10) for easy interpretability. It covers the entire range of depression severity assessed by established instruments. The metric allows...

  12. Mouse lung adhesion assay for Bordetella pertussis

    International Nuclear Information System (INIS)

    Burns, K.A.; Freer, J.H.

    1982-01-01

    The ability of Bordetella pertussis to adhere to cell surfaces has been demonstrated by adhesion to tissue culture cells and adhesion to chicken, hamster or rabbit trachea in organ culture. In this report a mouse lung assay for adhesion is described and the results obtained using two virulent strains of B. pertussis and their avirulent counterparts. These were a C modulation of one of the original virulent strains and a phase IV variant of the other virulent strain. Organisms were radiolabelled by adding 1 μCi (37 K Bq) of [ 14 C]glutamic acid per 10 ml of culture medium before inoculation and incubation for 5 days. The lungs were washed by perfusion in situ with at least two volumes (1 ml) of sterile 1% (w/v) casamino acids. The percentage of the inoculated organisms retained in the lungs was determined, after removal of the lungs, by one of the following two methods: viable count or radioactive count. Results for both methods were expressed as the percentage of the inoculum retained in the lungs plus or minus one standard deviation. (Auth.)

  13. A dot hybridization assay for the diagnosis of bacterial keratitis

    Science.gov (United States)

    Fang, Po-Chiung; Chien, Chun-Chih; Yu, Hun-Ju; Ho, Ren-Wen; Tseng, Shin-Ling; Lai, Yu-Hsuan

    2017-01-01

    Purpose To evaluate a bacterial dot hybridization (BDH) assay for the diagnosis of bacterial keratitis (BK). Methods Sixty-one qualified corneal scrapings from 61 patients with suspected microbial keratitis were collected consecutively and prospectively. Among the 61 patients, 16 cases were BK and 45 cases were non-BK, including fungal keratitis, viral keratitis, parasitic keratitis, and non-microbial keratitis. Molecular diagnosis of BK in these corneal scrapes was performed using the BDH assay with three universal bacterial probes (PB1, PB2, and PB3) and three genus-specific probes (Aci, Klb, and Psu) to detect Acinetobacter, Klebsiella, and Pseudomonas, respectively. Signals were standardized after grayscale image transformation for objective validation using receiver operating characteristic (ROC) curves. Results The standardized intensities for the three universal probes differed statistically significantly between the BK group and the non-BK group. Based on the ROC curves, the sensitivities of PB1, PB2, and PB3 were 81.3%, 81.3%, and 93.8%, and the specificities were 71.1%, 88.9%, and 91.1%, respectively. The sensitivity and specificity of the Psu probe were 92% and 100%, respectively, while those of the Aci and Klb probes could not be estimated because there were no BK cases caused by Acinetobacter spp. or Klebsiella spp. Conclusions The BDH assay is an effective molecular approach to improve the diagnosis of BK. Because the bias from bacterial contamination on the ocular surface can be minimized with signal standardization, the assay has the potential to be adopted for routine clinical practice. PMID:28484310

  14. After TGN1412: recent developments in cytokine release assays.

    Science.gov (United States)

    Stebbings, R; Eastwood, D; Poole, S; Thorpe, R

    2013-01-01

    The failure of regulatory science to keep pace with and support the development of new biological medicines was very publically highlighted in March 2006 when the first-in-man Phase I clinical trial of the immunomodulatory CD28-specific monoclonal antibody (mAb) TGN1412 ended in disaster when all six volunteers suffered a life-threatening adverse reaction termed a 'Cytokine Storm'. The poor predictive value of standard pre-clinical safety tests and animal models applied to TGN1412 demonstrated the need for a new generation of immunotoxicity assays and animal models that are both sensitive and predictive of clinical outcome in man. The non-predictive result obtained from pre-clinical safety testing in cynomolgus macaques has now been attributed to a lack of CD28 expression on CD4+ effector memory T-cells that therefore cannot be stimulated by TGN1412. In contrast, high levels of CD28 are expressed on human CD4+ effector memory T-cells, the source of most TGN1412-stimulated pro-inflammatory cytokines. Standard in vitro safety tests with human cells were also non-predictive as they did not replicate in vivo presentation of TGN1412. It was subsequently shown that, if an immobilized therapeutic mAb-based assay or endothelial cell co-culture assay was used to evaluate TGN1412, then these would have predicted a pro-inflammatory response in man. New in vitro assays based on these approaches are now being applied to emerging therapeutics to hopefully prevent a repeat of the TGN1412 incident. It has emerged that the mechanism of pro-inflammatory cytokine release stimulated by TGN1412 is different to that of other therapeutic mAbs, such that standard pro-inflammatory markers such as TNFα and IL-8 are not discriminatory. Rather, IL-2 release and lymphoproliferation are optimal readouts of a TGN1412-like pro-inflammatory response.

  15. Cell titration assay for measuring blastogenesis of bovine lymphocytes.

    Science.gov (United States)

    Baldwin, C L; Antczak, D F; Winter, A J

    1985-08-01

    The blastogenic response of bovine peripheral blood lymphocytes to phytohemagglutinin (PHA) and to microbial antigens was measured using a lymphocyte titration assay. Culture conditions, including lymphocyte concentrations, incubation periods and medium formulation, were established which produced linear or nearly linear responses over a range of cell concentrations. These conditions were established by testing lymphocytes from unimmunized cattle and from heifers infected with Brucella abortus with PHA and a B. abortus extract. Four cell concentrations in 2-fold increments were selected for measuring responses to PHA (3.125 X 10(3) to 2.5 X 10(4) cells/well) and to antigens (5.0 X 10(4) to 4.0 X 10(5) cells/well). The strength of response varied among animals and also over time for individual animals, but the titration assay allowed exponential proliferation to be distinguished from decline, which may have been due to overcrowding of microtiter wells, exhaustion of nutrients or induction of regulatory events. This assay provided a more reliable and discriminating method of evaluating lymphocyte proliferation responses than that achieved by single point assays. The displacement of the titration curves could be used to estimate the relative frequency of lymphocytes responding to antigens or mitogens.

  16. Expert system for transuranic waste assay

    International Nuclear Information System (INIS)

    Zoolalian, M.L.; Gibbs, A.; Kuhns, J.D.

    1989-01-01

    Transuranic wastes are generated at the Savannah River Site (SRS) as a result of routine production of nuclear materials. These wastes contain Pu-238 and Pu-239 and are placed into lined 55-gallon waste drums. The drums are placed on monitored storage pads pending shipment to the Waste Isolation Pilot Plant in New Mexico. A passive-active neutron (PAN) assay system is used to determine the mass of the radioactive material within the waste drums. Assay results are used to classify the wastes as either low-level or transuranic (TRU). During assays, the PAN assay system communicates with an IBM-AT computer. A Fortran computer program, called NEUT, controls and performs all data analyses. Unassisted, the NEUT program cannot adequately interpret assay results. To eliminate this limitation, an expert system shell was used to write a new algorithm, called the Transuranic Expert System (TRUX), to drive the NEUT program and add decision making capabilities for analysis of the assay results. The TRUX knowledge base was formulated by consulting with human experts in the field of neutron assay, by direct experimentation on the PAN assay system, and by observing operations on a daily basis. TRUX, with its improved ability to interpret assay results, has eliminated the need for close supervision by a human expert, allowing skilled technicians to operate the PAN assay system. 4 refs., 1 fig., 4 tabs

  17. The Salmonella Mutagenicity Assay: The Stethoscope of Genetic Toxicology for the 21st Century

    Science.gov (United States)

    Claxton, Larry D.; de A. Umbuzeiro, Gisela; DeMarini, David M.

    2010-01-01

    Objectives According to the 2007 National Research Council report Toxicology for the Twenty-First Century, modern methods (e.g., “omics,” in vitro assays, high-throughput testing, computational methods) will lead to the emergence of a new approach to toxicology. The Salmonella mammalian microsome mutagenicity assay has been central to the field of genetic toxicology since the 1970s. Here we document the paradigm shifts engendered by the assay, the validation and applications of the assay, and how the assay is a model for future in vitro toxicology assays. Data sources We searched PubMed, Scopus, and Web of Knowledge using key words relevant to the Salmonella assay and additional genotoxicity assays. Data extraction We merged the citations, removing duplicates, and categorized the papers by year and topic. Data synthesis The Salmonella assay led to two paradigm shifts: that some carcinogens were mutagens and that some environmental samples (e.g., air, water, soil, food, combustion emissions) were mutagenic. Although there are > 10,000 publications on the Salmonella assay, covering tens of thousands of agents, data on even more agents probably exist in unpublished form, largely as proprietary studies by industry. The Salmonella assay is a model for the development of 21st century in vitro toxicology assays in terms of the establishment of standard procedures, ability to test various agents, transferability across laboratories, validation and testing, and structure–activity analysis. Conclusions Similar to a stethoscope as a first-line, inexpensive tool in medicine, the Salmonella assay can serve a similar, indispensable role in the foreseeable future of 21st century toxicology. PMID:20682480

  18. Clinical impact of prostate specific antigen (PSA) inter-assay variability on management of prostate cancer.

    Science.gov (United States)

    Murthy, Vedang; Rishi, Anupam; Gupta, Sanjeev; Kannan, Sadhana; Mahantshetty, Umesh; Tongaonkar, Hemant; Bakshi, Ganesh; Prabhash, Kumar; Bhanushali, Paresh; Shinde, Bhoopal; Inamdar, Nitin; Shrivastava, Shyamkishore

    2016-01-01

    To evaluate the inter-assay variability of six commercially available prostate specific antigen (PSA) assays, its clinical impact in prostate cancer (PCa) and comparison of automated versus manual assays. Sera from 495 patients (425 with PCa and 70 men with Benign Prostatic Hyperplasia (BPH), were measured with six different assays [three automated assays (a-PSA) and three manual ELISA based assay (m-PSA)]. Variability, agreement and bias were measured and compared among assays using Bland Altman plots and Passing and Bablok regression analysis. The possible impact of inter-assay variability on important clinical scenarios was also studied. All the assays were well correlated (r: 0.88-0.98); however there was significant disagreement and bias between the systems, which were more pronounced among the a-PSA assays. The Bland Altman plot showed that the variability was high between the m-PSA assays and the standard Abbott system with mean difference of 3.8-5.8ng/ml. In contrast, the a-PSA had better agreement with mean difference of 0.8-2.3ng/ml. Beckman Coulter showed the best agreement to the institutional reference (slope-1.097; 95% CI: 1.06-1.14; p<0.05, and intercept-0.20; 95% CI-0.38-0.58; p<0.05, Passing Bablok). It led to significant variability in PCa risk stratification and failure to detect biochemical failure in more than 50% cases. The discrepancies between the assays lead to significant clinical misinterpretation with risk group migration and detection of biochemical failure post radiotherapy. There are significant discordances between automated and ELISA based assays. Copyright © 2015 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

  19. A Fully Automated High-Throughput Zebrafish Behavioral Ototoxicity Assay.

    Science.gov (United States)

    Todd, Douglas W; Philip, Rohit C; Niihori, Maki; Ringle, Ryan A; Coyle, Kelsey R; Zehri, Sobia F; Zabala, Leanne; Mudery, Jordan A; Francis, Ross H; Rodriguez, Jeffrey J; Jacob, Abraham

    2017-08-01

    Zebrafish animal models lend themselves to behavioral assays that can facilitate rapid screening of ototoxic, otoprotective, and otoregenerative drugs. Structurally similar to human inner ear hair cells, the mechanosensory hair cells on their lateral line allow the zebrafish to sense water flow and orient head-to-current in a behavior called rheotaxis. This rheotaxis behavior deteriorates in a dose-dependent manner with increased exposure to the ototoxin cisplatin, thereby establishing itself as an excellent biomarker for anatomic damage to lateral line hair cells. Building on work by our group and others, we have built a new, fully automated high-throughput behavioral assay system that uses automated image analysis techniques to quantify rheotaxis behavior. This novel system consists of a custom-designed swimming apparatus and imaging system consisting of network-controlled Raspberry Pi microcomputers capturing infrared video. Automated analysis techniques detect individual zebrafish, compute their orientation, and quantify the rheotaxis behavior of a zebrafish test population, producing a powerful, high-throughput behavioral assay. Using our fully automated biological assay to test a standardized ototoxic dose of cisplatin against varying doses of compounds that protect or regenerate hair cells may facilitate rapid translation of candidate drugs into preclinical mammalian models of hearing loss.

  20. Assay development status report for total cyanide

    Energy Technology Data Exchange (ETDEWEB)

    Simpson, B.C. [Westinghouse Hanford Co., Richland, WA (United States); Jones, T.E.; Pool, K.H. [Pacific Northwest Lab., Richland, WA (United States)

    1993-02-01

    A validated cyanide assay that is applicable to a variety of tank waste matrices is necessary to resolve certain waste tank safety issues and for purposes of overall waste characterization. The target for this effort is an assay with an applicable range of greater than 1,000 ppM (0.10 wt%) total cyanide and a confidence level greater than 80%. Figure 1 illustrates the operating regime of the proposed cyanide assay method. The Assay Development Status Report for Total Cyanide will summarize the past experience with cyanide analyses on-tank waste matrices and will rate the status of the analytical methods used to assay total cyanide (CN{sup {minus}} ion) in the tank waste matrices as acceptable or unacceptable. This paper will also briefly describe the current efforts for improving analytical resolution of the assays and the attempts at speciation.

  1. Improved curve-fitting, parallelism testing, characterisation of sensitivity and specificity, validation, and optimization for radioligand assays

    International Nuclear Information System (INIS)

    Rodbard, D.; Munson, P.J.; DeLean, A.

    1977-01-01

    A new computer program has been developed to simultaneously 'fit' a family of curves, based on the four parameter logistic equation. This provides an optimal test of 'parallelism', permits estimation of shared parameters based on all of the available data, and provides 'combined potency estimates'. Several simple general methods have been developed to permit testing of 'parallelism' of standard and unknown preparations, irrespective of the method used for dose interpolation. The use of the mass-action law equations to describe RIA dose response curves and for dose interpolation, has been extented to apply to cases involving up to three independant classes of sites (one 'non-saturable'), corresponding to a 5 parameter Scatchard plot model. We discuss a statistically meaningful definition of the minimal detectable dose in an assay. Simple computer programs have been developed for analysis of the typical 'recovery' experiment, i.e. when graded doses of unlabeled ligand are added to serum, to test the validity of the assay. A new program has been developed, to provide least squares estimation of parameters for the association and dissociation kinetics of RIA and radioreceptor assay systems, based on simlutaneous analysis of all relevant data. Improved programs have been developed for optimization of conditions of RIAs employing delayed addition of labeled ligand. (orig./AJ) [de

  2. Development of a rapid ferricyanide mediated assay for biochemical oxygen demand using a mixed microbial consortium.

    Science.gov (United States)

    Catterall, Kylie; Zhao, Huijun; Pasco, Neil; John, Richard

    2003-06-01

    Ferricyanide-mediated (FM) microbial reactions were used for the rapid determination of the biochemical oxygen demand (BOD) of a range of synthetic and real wastewater samples. Four single-species microbial seeds and a synthetically prepared microbial consortium were compared. In all cases, the microbial consortium exhibited a greater extent and rate of biodegradation compared to the individual microbial seeds. Markedly improved correlation to the standard BOD5 method was also noted for the microbial consortium (compared to the single-species seeds). A linear dynamic range up to 200 mg BOD5 L(-1) was observed, which is considerably greater than the linear range of the standard BOD5 assay and most other rapid BOD assays reported. In addition, biodegradation efficiencies comparable to the 5-day BOD5 assay (and much greater than other rapid BOD assays) were observed in 3 h. A highly significant correlation (R = 0.935, p = 0.000, n = 30) between the FM-BOD method and the standard BOD5 method was found for a wide diversity of real wastewater samples. The results indicate that the FM-BOD assay is a promising, rapid, alternative to the standard 5-day BOD5 assay.

  3. New automated pellet/powder assay system

    International Nuclear Information System (INIS)

    Olsen, R.N.

    1975-01-01

    This paper discusses an automated, high precision, pellet/ powder assay system. The system is an active assay system using a small isotopic neutron source and a coincidence detection system. The handling of the pellet powder samples has been automated and a programmable calculator has been integrated into the system to provide control and data analysis. The versatile system can assay uranium or plutonium in either active or passive modes

  4. Development of Plaque Assay Systems for Poliovirus.

    Science.gov (United States)

    1982-04-01

    inter- action must be ascertained for each type of virus to be collected and assayed. The vaccine strain of poliovirus type 1 (Sabin) was chosen as a...1067 DEVELOPMENT OF PLAQUE ASSAY SYSTEMS FOR POLIOVIRUS (U) by R.E. Fulton and K. Munroe Abstract During the summer months of 1978, Ms. Krista Munroe...quantitation of infectious poliovirus type 1. .Two different plaque assay techniques were developed and compared. The results of this work are presented

  5. A lateral electrophoretic flow diagnostic assay

    OpenAIRE

    Lin, R; Skandarajah, A; Gerver, RE; Neira, HD; Fletcher, DA; Herr, AE

    2015-01-01

    © 2015 The Royal Society of Chemistry. Immunochromatographic assays are a cornerstone tool in disease screening. To complement existing lateral flow assays (based on wicking flow) we introduce a lateral flow format that employs directed electrophoretic transport. The format is termed a "lateral e-flow assay" and is designed to support multiplexed detection using immobilized reaction volumes of capture antigen. To fabricate the lateral e-flow device, we employ mask-based UV photopatterning to ...

  6. Development of in vitro assay method with radioisotope

    International Nuclear Information System (INIS)

    Choi, Chang Woon; Lim, S. M.; An, S. H.; Woo, K. S.; Chung, W. S.; Lim, S. J.; Hong, S. W.; Oh, O. D.

    1999-04-01

    Radioimmunoassay (RIA) and related competitive protein-binding methods began a little over 20 years ago as a cumbersome research methodology in a few specialized laboratories. Endocrinology has been greatly enriched by the new knowledge that has come as a direct result of RIA methods. Establishment of the taxol RIA system will be expected to develop RIA for drug monitoring. Scintillation proximity assay was useful since any separation step is not required, it has the advantage of dealing with multiple samples. The increased sensitivity of the new assay in determining HCV RT([ 125 I]dUTP) suggests that it would be worth investigating whether the system can be applied to analysis. [ 125 I] lodotyramine with 98.5% radiochemical purity. Optimal background counts was certificated using varied radioactivity of radionuclides. Appropriate standard curve was obtained from SPA method successively, and the concentration of hCG from unknown serum was determined by standard curve. The result concentration of hCG from unknown serum was determined by synthesized successively and purified by HPLC system. Hybridoma reducing monoclonal anti thyroglobulin antibodies titer is measured by ELISA. These studies play an important role in development of in vitro assay with radionuclides

  7. Development of in vitro assay method with radioisotope

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Chang Woon; Lim, S. M.; An, S. H.; Woo, K. S.; Chung, W. S.; Lim, S. J.; Hong, S. W. [Korea Atomic Energy Research Institute. Korea Cancer Center Hospital, Seoul (Korea, Republic of); Oh, O. D. [Yonsei University, Seoul (Korea, Republic of)

    1999-04-01

    Radioimmunoassay (RIA) and related competitive protein-binding methods began a little over 20 years ago as a cumbersome research methodology in a few specialized laboratories. Endocrinology has been greatly enriched by the new knowledge that has come as a direct result of RIA methods. Establishment of the taxol RIA system will be expected to develop RIA for drug monitoring. Scintillation proximity assay was useful since any separation step is not required, it has the advantage of dealing with multiple samples. The increased sensitivity of the new assay in determining HCV RT([{sup 125}I]dUTP) suggests that it would be worth investigating whether the system can be applied to analysis. [{sup 125}I] lodotyramine with 98.5% radiochemical purity. Optimal background counts was certificated using varied radioactivity of radionuclides. Appropriate standard curve was obtained from SPA method successively, and the concentration of hCG from unknown serum was determined by standard curve. The result concentration of hCG from unknown serum was determined by synthesized successively and purified by HPLC system. Hybridoma reducing monoclonal anti thyroglobulin antibodies titer is measured by ELISA. These studies play an important role in development of in vitro assay with radionuclides.

  8. Ecogenotoxicity testing of aquatic environment by comet assay in plants

    Directory of Open Access Journals (Sweden)

    Anita Mukherjee

    2015-05-01

    Full Text Available One of the goals of environmental monitoring is the detection of potentially hazardous compounds in water. We have set up a standard method to apply the Comet assay in aquatic plants that could be of great interest to evaluate cytotoxicity, genotoxicity and oxidative stress on the same species regarded as most sensitive to environmental pollutants. The aim of the present study was to set up of standardized procedure to evaluate genotoxicity in aquatic plants- Ceratophyllum demersum one that is submerged free floating and the other is Lemna minor - a fresh water floating plant by Comet assay. Electrophoresis and unwinding times were adapted to obtain minimum DNA migration evaluated as tail intensity % or tail moment in the control group and, at the same time maximum sensitivity for DNA damage with known genotoxicants. We further investigated the cytotoxicity and oxidative stress induced in the same species. Based on the repeatability of results obtained we suggest that Ceratophyllum, Lemna can serve as model species and Comet assay could be adopted to monitor the eco-genotoxicity of water pollutants.

  9. Development of Best practices document for Peptide Standards | Office of Cancer Clinical Proteomics Research

    Science.gov (United States)

    The Assay Development Working Group (ADWG) of the CPTAC Program is currently drafting a document to propose best practices for generation, quantification, storage, and handling of peptide standards used for mass spectrometry-based assays, as well as interpretation of quantitative proteomic data based on peptide standards. The ADWG is seeking input from commercial entities that provide peptide standards for mass spectrometry-based assays or that perform amino acid analysis.

  10. Matrix effects of TRU [transuranic] assays using the SWEPP PAN assay system

    International Nuclear Information System (INIS)

    Smith, J.R.

    1990-08-01

    The Drum Assay System (DAS) at the Stored Waste Experimental Pilot Plant (SWEPP) is a second-generation active-passive neutron assay system. It has been used to assay over 5000 208-liter drums of transuranic waste from the Rocky Flats Plant (RFP). Data from these assays have been examined and compared with the assays performed at Rocky Flats, mainly utilize counting of 239 Pu gamma rays. For the most part the passive assays are in very good agreement with the Rocky Flats assays. The active assays are strongly correlated with the results of the other two methods, but require matrix-dependent correction factors beyond those provided by the system itself. A set of matrix-dependent correction factors has been developed from the study of the assay results. 3 refs., 4 figs., 3 tabs

  11. Safeguards and Non-destructive Assay

    International Nuclear Information System (INIS)

    Carchon, R.; Bruggeman, M.

    2001-01-01

    SCK-CEN's programme on safeguards and non-destructive assay includes: (1) various activities to assure nuclear materials accountancy; (2) contributes to the implementation of Integrated Safeguards measures in Belgium and to assist the IAEA through the Belgian Support Programme; (3) renders services to internal and external customers in the field of safeguards; (4) improves passive neutron coincidence counting techniques for waste assay and safeguards verification measurements by R and D on correlation algorithms implemented via software or dedicated hardware; (5) improves gamma assay techniques for waste assay by implementing advanced scanning techniques and different correlation algorithms; and (6) develops numerical calibration techniques. Major achievements in these areas in 2000 are reported

  12. Proximity assays for sensitive quantification of proteins.

    Science.gov (United States)

    Greenwood, Christina; Ruff, David; Kirvell, Sara; Johnson, Gemma; Dhillon, Harvinder S; Bustin, Stephen A

    2015-06-01

    Proximity assays are immunohistochemical tools that utilise two or more DNA-tagged aptamers or antibodies binding in close proximity to the same protein or protein complex. Amplification by PCR or isothermal methods and hybridisation of a labelled probe to its DNA target generates a signal that enables sensitive and robust detection of proteins, protein modifications or protein-protein interactions. Assays can be carried out in homogeneous or solid phase formats and in situ assays can visualise single protein molecules or complexes with high spatial accuracy. These properties highlight the potential of proximity assays in research, diagnostic, pharmacological and many other applications that require sensitive, specific and accurate assessments of protein expression.

  13. Thyroglobulin measurement by highly sensitive assays

    DEFF Research Database (Denmark)

    Giovanella, Luca; Feldt-Rasmussen, Ulla; Verburg, Frederik A

    2015-01-01

    of thyroglobulin (Tg), circulating Tg serves as a biochemical marker of persistent or recurrent disease in the follow-up of DTC. Due to the suboptimal clinical detection rate of older Tg assays endogenous or exogenous thyrotropin (TSH) stimulations are recommended for unmasking occult disease. However...... in current clinical guidelines, such assays are now increasingly used in clinical practice. As serum Tg measurement is a technically challenging assay and criteria to define a 'highly sensitive' assay may be different, a good knowledge of the technical difficulties and interpretation criteria is of paramount...

  14. 233U Assay A Neutron NDA System

    Energy Technology Data Exchange (ETDEWEB)

    Hensley, D.C.; Lucero, A.J.; Pierce, L.

    1998-11-17

    The assay of highly enriched {sup 233}U material presents some unique challenges. Techniques which apply to the assay of materials of Pu or enriched {sup 235}U do not convert easily over to the assay of {sup 233}U. A specialized neutron assay device is being fabricated to exploit the singles neutron signal, the weak correlated neutron signal, and an active correlated signal. These pieces of information when combined with {gamma} ray isotopics information should give a good overall determination of {sup 233}U material now stored in bldg. 3019 at the Oak Ridge National Laboratory.

  15. Parameter estimation and use of gamma interferon assay for the diagnosis of bovine tuberculosis in Brazil

    Directory of Open Access Journals (Sweden)

    Luciano B. Lopes

    2012-04-01

    Full Text Available This study aimed to evaluate the interference of tuberculin test on the gamma-interferon (INFg assay, to estimate the sensitivity and specificity of the INFg assay in Brazilian